phytoestrogens and Disease-Models--Animal

A wide range of health benefits have been ascribed to soya intake including a lowered risk of osteoporosis, heart disease, breast cancer, and menopausal symptoms. Because it is a hormonally active diet, however, soya can also be endocrine disrupting, suggesting that intake has the potential to cause adverse health effects in certain circumstances, particularly when exposure occurs during development. Consequently, the question of whether or not soya phyto-oestrogens are beneficial or harmful to human health is neither straightforward nor universally applicable to all groups. Possible benefits and risks depend on age, health status, and even the presence or absence of specific gut microflora. As global consumption increases, greater awareness and consideration of the endocrine-disrupting properties of soya by nutrition specialists and other health practitioners is needed. Consumption by infants and small children is of particular concern because their hormone-sensitive organs, including the brain and reproductive system, are still undergoing sexual differentiation and maturation. Thus, their susceptibility to the endocrine-disrupting activities of soya phyto-oestrogens may be especially high. As oestrogen receptor partial agonists with molecular and cellular properties similar to anthropogenic endocrine disruptors such as bisphenol A, the soya phyto-oestrogens provide an interesting model for how attitudes about what is 'synthetic' v. what is 'natural,' shapes understanding and perception of what it means for a compound to be endocrine disrupting and/or potentially harmful. This review describes the endocrine-disrupting properties of soya phyto-oestrogens with a focus on neuroendocrine development and behaviour.

Topics: Animals; Benzhydryl Compounds; Diet; Disease Models, Animal; Endocrine Disruptors; Glycine max; Humans; Isoflavones; Neurosecretory Systems; Phenols; Phytoestrogens; Reproduction; Sexual Behavior; Sexual Behavior, Animal

Phytoestrogens are polyphenols similar to human estrogens found in plants or derived from plant precursors. Phytoestrogens are found in high concentration in soya, flaxseed and other seeds, fruits, vegetables, cereals, tea, chocolate, etc. They comprise several classes of chemical compounds (stilbenes, coumestans, isoflavones, ellagitannins, and lignans) which are structurally similar to endogenous estrogens but which can have both estrogenic and antiestrogenic effects. Although epidemiological and experimental evidence indicates that intake of phytoestrogens in foods may be protective against certain chronic diseases, discrepancies have been observed between in vivo and in vitro experiments. The microbial transformations have not been reported so far in stilbenes and coumestans. However, isoflavones, ellagitanins, and lignans are metabolized by intestinal bacteria to produce equol, urolithins, and enterolignans, respectively. Equol, urolithin, and enterolignans are more bioavailable, and have more estrogenic/antiestrogenic and antioxidant activity than their precursors. Moreover, equol, urolithins and enterolignans have anti-inflammatory effects and induce antiproliferative and apoptosis-inducing activities. The transformation of isoflavones, ellagitanins, and lignans by intestinal microbiota is essential to be protective against certain chronic diseases, as cancer, cardiovascular disease, osteoporosis, and menopausal symptoms. Bioavailability, bioactivity, and health effects of dietary phytoestrogens are strongly determined by the intestinal bacteria of each individual.

Topics: Animals; Chocolate; Chronic Disease; Coumarins; Disease Models, Animal; Edible Grain; Flax; Fruit; Gastrointestinal Microbiome; Glycine max; Humans; Hydrolyzable Tannins; Intestines; Isoflavones; Lignans; Phytoestrogens; Polyphenols; Stilbenes; Tea; Vegetables

Isoflavones are the most widely consumed phytoestrogens. Besides being a dietary constituent, their consumption has been increasing in the form of herbal supplements and as promising alternatives to hormonal replacement therapy, in conjunction with prescription medicines. Isoflavones are extensively metabolized by phase I and II enzymes and are substrates of drug transporters. At high concentrations isoflavones may interact with drug metabolizing enzymes and drug transporters and modulate their activity, thus, altering the absorption, metabolism, distribution, excretion and toxicity profile of the co-administered drugs. This review summarizes the up-to-date literature of isoflavone-drug interactions giving insight into the possible mechanisms of interactions, in vitro-in vivo correlation and their implications on clinical outcomes.

Topics: Animals; Cytochrome P-450 Enzyme System; Disease Models, Animal; Herb-Drug Interactions; Humans; Inactivation, Metabolic; Isoflavones; Phytoestrogens; Phytotherapy; Plants, Medicinal; Prescription Drugs

Some studies have considered the association between diet and uterine fibroid risk, but the issue is largely unsettled. To identify potential modifiable risk factors for fibroid development, we have herein systematically reviewed prior publications dealing with this aspect. Comprehensive searches in electronic databases were conducted to collect studies published on association between uterine leiomyomas and both nutrients and food groups. We identified 13 publications deriving from 4 case-control, 3 cross-sectional, and 4 cohort studies. A protective effect has been demonstrated for consumption of fruits and green vegetables in both case-control and cohort studies. Moreover, very recent cross-sectional and case-control studies evaluating serum levels of 25-hydroxyvitamin-D3 tend to indicate that vitamin D insufficiency, which may in part be due to the diet intake, may play an important role in the development of uterine fibroids. No association was found with the intake of fibers, vitamin C and E, phytoestrogens and carotenoids, whereas association was controversial for the consumption of meat, fish, dairy products, and vitamin A. Most data have also been discussed herein in light of the available experimental and animal model results. These findings may be useful in devising nutritional strategies to reduce leiomyoma risk in humans.

Topics: Animals; Ascorbic Acid; Carotenoids; Dairy Products; Diet; Dietary Fats; Dietary Fiber; Disease Models, Animal; Folic Acid; Fruit; Humans; Leiomyoma; Meat; Phytoestrogens; Risk Factors; Seafood; Soy Foods; Vegetables; Vitamin A; Vitamin D; Vitamin E

Flaxseed (FS), rich in the phytoestrogen lignans and α-linolenic acid-rich oil, has been suggested to have an anticancer effect. Questions remain whether FS and its lignan and oil components are effective in reducing breast cancer risk and tumour growth, and can interact beneficially with breast cancer drugs. To find answers, in vitro, animal, observational, and clinical studies on FS and its lignan and oil components were reviewed. The majority of studies in various rodent models show that 2.5%-10% FS diet or the equivalent amount of lignan or oil reduces tumour growth. Ten percent FS and equivalent lignans do not interfere with but rather increase the effectiveness of tamoxifen (80 mg/day) while the 4% FS oil increases trastuzumab/Herceptin (2.5 mg/kg) effectiveness. Observational studies show that FS and lignan intake, urinary excretion, or serum levels are associated with reduced risk, particularly in postmenopausal women. Lignans reduce breast cancer and all-cause mortality by 33%-70% and 40%-53%, respectively, without reducing tamoxifen effectiveness. Clinical trials show that FS (25 g/day with 50 mg lignans; 32 days) reduces tumour growth in breast cancer patients and lignans (50 mg/day; 1 year) reduces risk in premenopausal women. Mechanisms include decreased cell proliferation and angiogenesis and increased apoptosis through modulation of estrogen metabolism and estrogen receptor and growth factor receptor signalling pathways. More clinical trials are needed but current overall evidence indicates that FS and its components are effective in the risk reduction and treatment of breast cancer and safe for consumption by breast cancer patients.

Topics: alpha-Linolenic Acid; Animals; Antineoplastic Agents; Breast Neoplasms; Disease Models, Animal; Drug Interactions; Female; Flax; Humans; Lignans; Phytoestrogens; Risk Factors

Orange juice (OJ) is among the most consumed fruit juices worldwide, and its chemopreventive action is fairly addressed in the literature. This review critically presents the available evidence linking OJ with cancer chemoprevention and on discussing the putative mechanisms and negative health effects. The chemopreventive action of OJ is related to its effect on metabolic enzymes and its antiinflammatory, cytoprotective/apoptotic, hormonal, cell signaling-modulating, antioxidant, and antigenotoxic effects. Most studies on OJ are in vitro, and few are conducted in vivo. Results from in vitro studies must be interpreted carefully because these findings do not consider in vivo bioavailability. However, such results are useful for studying the impact of different processing and storage methods on OJ's chemopreventive effect. Evidence of OJ's chemoprevention in humans is limited. OJ is antimutagenic in bacteria and antigenotoxic in humans and rodents. Studies using rodent cancer models showed that OJ is cancer chemopreventive, influencing either the induction stage or the promotion stage. The composition and, therefore, the chemopreventive action of OJ might be influenced by different cultivars, climates, extraction methods, packaging, storage temperatures, and shelf lives, among other factors. Epidemiological studies and randomized controlled intervention studies in humans evaluating the chemopreventive effect of OJ, taking into consideration variability in OJ composition, are needed.

Topics: Animals; Anti-Infective Agents; Anti-Inflammatory Agents; Antioxidants; Beverages; Chemoprevention; Citrus; Cytoprotection; Disease Models, Animal; DNA Damage; Fruit; Humans; Neoplasms; Phytoestrogens; Randomized Controlled Trials as Topic

Developmental exposure to estrogenic compounds can disrupt sexual differentiation and adult reproductive function in many animals including humans. Phytoestrogens (plant estrogens) in the diet comprise a significant source of estrogenic exposure to humans, particularly in infants who are fed soy-based infant formula. Animal models have been developed to test the effects of phytoestrogen exposure on the developing fetus and neonate. Here we review studies quantifying the amount of phytoestrogen exposure in human adults and infants and discuss the few available epidemiological studies that have addressed long-term consequences of developmental phytoestrogen exposure. We then describe in detail rodent models of developmental exposure to the most prevalent phytoestrogen in soy products, genistein, and the effects of this exposure on female reproductive function. These models have used various dosing strategies to mimic the phytoestrogen levels in human populations. Serum circulating levels of genistein following each of the models and their correlation to reproductive outcomes are also discussed. Taken together, the studies clearly demonstrate that environmentally relevant doses of genistein have significant negative impacts on ovarian differentiation, estrous cyclicity, and fertility in the rodent model. Additional studies of reproductive function in human populations exposed to high levels of phytoestrogens during development are warranted.

Topics: Adult; Animals; Animals, Newborn; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Administration Routes; Environmental Exposure; Estrous Cycle; Female; Fertility; Genistein; Humans; Infant; Infant, Newborn; Isoflavones; Male; Maternal Exposure; Organ Size; Ovary; Phytoestrogens; Pregnancy; Prenatal Exposure Delayed Effects; Uterus

A high intake of fruits and vegetables is associated with a lower risk of cancer. In this context, considerable attention is paid to Asian populations who consume high amounts of soy and soy-derived isoflavones, and have a lower risk for several cancer types such as breast and prostate cancers than populations in Western countries. Hence, interest focuses on soyfoods, soy products, and soy ingredients such as isoflavones with regard to their possible beneficial effects that were observed in numerous experiments and studies. The outcomes of the studies are not always conclusive, are often contradictory depending on the experimental conditions, and are, therefore, difficult to interpret. Isoflavone research revealed not only beneficial but also adverse effects, for instance, on the reproductive system. This is also the case with tumor-promoting effects on, for example, breast tissue. Isoflavone extracts and supplements are often used for the treatment of menopausal symptoms and for the prevention of age-associated conditions such as cardiovascular diseases and osteoporosis in postmenopausal women. In relation to this, questions about the effectiveness and safety of isoflavones have to be clarified. Moreover, there are concerns about the maternal consumption of isoflavones due to the development of leukemia in infants. In contrast, men may benefit from the intake of isoflavones with regard to reducing the risk of prostate cancer. Therefore, this review examines the risks but also the benefits of isoflavones with regard to various kinds of cancer, which can be derived from animal and human studies as well as from in vitro experiments.

Topics: Animals; Anticarcinogenic Agents; Biological Availability; Carcinogenicity Tests; Carcinogens; Cardiovascular Diseases; Diet; Disease Models, Animal; Drug Combinations; Female; Glycine max; Hormone Replacement Therapy; Humans; Isoflavones; Male; Neoplasms; Osteoporosis, Postmenopausal; Phytoestrogens; Phytotherapy; Plant Extracts; Risk Assessment

Biological sex plays an important role in normal cardiac physiology as well as in the heart's response to cardiac disease. Women generally have better cardiac function and survival than do men in the face of cardiac disease; however, this sex difference is lost when comparing postmenopausal women with age-matched men. Animal models of cardiac disease mirror what is seen in humans. Sex steroid hormones contribute significantly to sex-based differences in cardiac disease outcomes. Estrogen is generally considered to be cardioprotective, whereas testosterone is thought to be detrimental to heart function. Environmental estrogen-like molecules, such as phytoestrogens, can also affect cardiac physiology in both a positive and a negative manner.

Topics: Animals; Disease Models, Animal; Estrogens; Female; Heart; Heart Diseases; Humans; Male; Mice; Phytoestrogens; Sex Characteristics; Testosterone

Isoflavones and their related flavonoid compounds exert antiviral properties in vitro and in vivo against a wide range of viruses. Genistein is, by far, the most studied soy isoflavone in this regard, and it has been shown to inhibit the infectivity of enveloped or nonenveloped viruses, as well as single-stranded or double-stranded RNA or DNA viruses. At concentrations ranging from physiological to supraphysiological (3.7-370 muM), flavonoids, including genistein, have been shown to reduce the infectivity of a variety of viruses affecting humans and animals, including adenovirus, herpes simplex virus, human immunodeficiency virus, porcine reproductive and respiratory syndrome virus, and rotavirus. Although the biological properties of the flavonoids are well studied, the mechanisms of action underlying their antiviral properties have not been fully elucidated. Current results suggest a combination of effects on both the virus and the host cell. Isoflavones have been reported to affect virus binding, entry, replication, viral protein translation and formation of certain virus envelope glycoprotein complexes. Isoflavones also affect a variety of host cell signaling processes, including induction of gene transcription factors and secretion of cytokines. The efficacy of isoflavones and related flavonoids in virus infectivity in in vitro bioassays is dependent on the dose, frequency of administration and combination of isoflavones used. Despite promising in vitro results, there is lack of data confirming the in vivo efficacy of soy isoflavones. Thus, investigations using appropriate in vivo virus infectivity models to examine pharmacological and especially physiological doses of flavonoids are warranted.

Topics: Animals; Anticarcinogenic Agents; Antiviral Agents; Biological Availability; Diet; Disease Models, Animal; Flavonoids; Genistein; Glycine max; Humans; Isoflavones; Phytoestrogens; Species Specificity; Virus Diseases; Viruses

Recent large clinical trials demonstrating deleterious effects of postmenopausal hormone replacement therapy have raised interest in the use of food products and extracts containing phytoestrogens as potential safe alternatives for menopausal symptoms, age-related cognitive decline and neurodegenerative diseases. While numerous preclinical studies and various clinical trials point to beneficial effects of estrogens on the brain, phytoestrogens from several sources share many of these estrogenic effects, in addition to having unique activities distinct from natural estrogens. Numerous in vitro and in vivo studies show potential neuroprotective properties of phytoestrogens on the brain in conditions ranging from aging to neurodegenerative disease and cerebral ischemia. Although dosage, timing and safety concerns remain to be addressed before their therapeutic use in human populations can be recommended, their safety profile and some intriguing studies on human cognition in aging suggest that further clinical study of these compounds for brain health is warranted.

Topics: Animals; Cell Line; Disease Models, Animal; Humans; Molecular Structure; Neurodegenerative Diseases; Neuroprotective Agents; Phytoestrogens

Due to their ability to mimic the actions of mammalian estrogens, soy phytoestrogens have been proposed as potential therapeutic agents to aid in preventing postmenopausal bone loss. In vitro, phytoestrogens promote osteoblastogenesis and inhibit osteoclastogenesis. Although a relatively large number of intervention studies have been undertaken in animals and humans, the efficacy of phytoestrogens as bone-protective agents in vivo remains unclear. Differences in the bioactivities of individual phytoestrogens, differences in phytoestrogen metabolism and bioavailability within different study populations, and imprecise reporting of the dose of phytoestrogens administered in intervention studies may have contributed to the disparity in study findings.

Topics: Animals; Bone and Bones; Bone Density Conservation Agents; Clinical Trials as Topic; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Glycine max; Humans; Middle Aged; Osteoporosis, Postmenopausal; Phytoestrogens

Phytoestrogens are non-steroidal estrogens widely distributed in many kinds of plants. They are natural compounds structurally similar to estrogen and with estrogenic or anti-androgenic activities. Prostate cancer (PCa) and benign prostatic hyperplasia (BPH) are androgen-dependent and associated with age. Recently, in many epidemiological and experimental researches, it has been reported that phytoestrogens play a role in the prevention and treatment of PCa and BPH. Regulation of sexual hormones, inhibition of cell proliferation, induction of cell apoptosis and anti-oxidation of such plant estrogens may be involved in the mechanisms.

Topics: Animals; Disease Models, Animal; Humans; Male; Phytoestrogens; Prostatic Hyperplasia; Prostatic Neoplasms; Rats; Rats, Sprague-Dawley; Rats, Wistar

The impact of soyfood intake on breast cancer risk has been investigated extensively. Much of this focus can be attributed to the soybean being a dietary source that is uniquely rich in isoflavones. The chemical structure of isoflavones is similar to that of estrogen, and isoflavones bind to both estrogen receptors (ER alpha and ER beta) (although they preferentially bind to and activate ER beta) and exert estrogen-like effects under some experimental conditions. Isoflavones also possess nonhormonal properties that are associated with the inhibition of cancer cell growth. Thus, there are several possible mechanisms by which soy may reduce the risk of breast cancer. However, the role of isoflavones in breast cancer has become controversial because, in contrast to the possible beneficial effects, some data from in vitro and animal studies suggest that isoflavones, especially genistein, the aglycone of the main soybean isoflavone genistin, may stimulate the growth of estrogen-sensitive tumors. Limited human data directly address the tumor-promoting effects of isoflavones and soy. Because the use of soyfoods and isoflavone supplements is increasing, it is important from a public health perspective to understand the impact of these products on breast cancer risk in women at high risk of the disease and on the survival of breast cancer patients. To this end, a workshop was held in November 2005 to review the existing literature and to make research recommendations. This paper summarizes the workshop findings and recommendations. The primary research recommendation is that the impact of isoflavones on breast tissue needs to be evaluated at the cellular level in women at high risk for breast cancer.

Topics: Animals; Anticarcinogenic Agents; Breast Neoplasms; Carcinogens; China; Clinical Trials as Topic; Congresses as Topic; Disease Models, Animal; Female; Genistein; Humans; Isoflavones; Los Angeles; Neoplasms, Hormone-Dependent; Phytoestrogens; Population Surveillance; Receptors, Estrogen; Risk Factors; Soy Foods

This review presents findings with clear statements from the literature as well as own results of effects of soy, red clover and their isoflavones as well as of the Cimicifuga racemosa extract BNO 1055. Experimental and clinical effects on climacteric complaints, osteoprotective effects, activity in the urogenital tract, and risks concerning cardiovascular diseases and mammary and endometrial tissue will be compared, also in comparison to classical hormone preparations. The question whether soy and red clover products and/or Cimicifuga racemosa (CR) preparations are endocrine disrupters or may fulfill the criteria of the so-called phyto-SERMs will be discussed.. Review of selected publications since 1980 and summary of unpublished own results of the authors.. Experimental and clinical evidences suggest that soy/red clover and their isoflavones do not fulfill the criteria of an ideal SERM. They appear to have mild osteoprotective effects but do not improve climacteric complaints. Furthermore, they seem to stimulate uterine growth and mammary epithelial proliferation. In ovariectomized rats, the CR extract BNO 1055 showed many of the beneficial effects of 17beta-estradiol, including effects in the brain/hypothalamus to reduce serum LH levels, effects in the bone to prevent osteoporosis and estrogenic effects in the urinary bladder. The CR extract BNO 1055 had no uterotrophic effect.. If clinical studies confirm these results, the Cimicifuga racemosa preparation BNO 1055 would appear as an ideal SERM and may therefore be an alternative to hormone replacement therapy.

Topics: Animals; Bone and Bones; Breast; Cimicifuga; Disease Models, Animal; Endocrine System; Endometrium; Estrogen Replacement Therapy; Estrogens, Non-Steroidal; Female; Glycine max; Hot Flashes; Humans; Isoflavones; Menopause; Phytoestrogens; Phytotherapy; Plant Extracts; Plant Preparations; Rats; Selective Estrogen Receptor Modulators; Trifolium; Urinary Bladder; Uterus

Approximately 50% of Americans use dietary supplements on a regular basis spending an estimated $20 billion on supplements in the year 2000. Soy contains genistein and daidzein, two phytoestrogens, which work through the estrogen receptor and cause alterations in serum lipids, bone metabolism, and possibly cognition. In this article, we review the issues regarding the interpretation with studies using soy-based isoflavones, discuss their mechanism of action, and review the literature on the effect of these bio-active compounds on lipid metabolism, osteoblasts and osteoclasts, bone markers, bone mineral density, and cognition.

Topics: Animals; Bone Density; Clinical Trials as Topic; Cognition; Dietary Supplements; Disease Models, Animal; Estrogens, Non-Steroidal; Female; Glycine max; Humans; Isoflavones; Menopause; Osteoporosis, Postmenopausal; Phytoestrogens; Phytotherapy; Plant Extracts; Plant Preparations; Rats

Androgcns are required to maintain the integrity of the prostate and the survival of androgen dependent epithelial cells within the gland. Anti-androgens arc the primary treatment strategy for non-localized prostate cancer, but ultimately fail over time with the development of androgen independent tumors. Estrogens affect the growth and development of the prostate and may affect the development of prostate cancer. Because of the side effects of estrogen treatment alternative therapies include the use of phytoestrogens as chemopreventative and chemotherapeutic treatment modalities. Phytoestrogens, can cause growth arrest and in some cases apoptosis in prostate cancer cells in vivo and in vitro. This may be due to the estrogenic properties of the compounds or alternative mechanisms of action. A number of phytoestrogens have been shown to have anti-androgenic effects and anti-oxidant activities. Other mechanisms include inhibition of 5alpha-reductase, 17beta-hydroxysteroid dehydrogenase, aromatase, tyrosine specific protein kinases and DNA topoisomerase II. This review examines the possible relation between phytoestrogens and prostate cancer and their possible use in prostate cancer prevention or management.

Topics: Animals; Anticarcinogenic Agents; Antineoplastic Agents, Phytogenic; Biological Availability; Disease Models, Animal; Estradiol; Estrogens, Non-Steroidal; Humans; Isoflavones; Male; Mice; Phytoestrogens; Plant Preparations; Prostatic Neoplasms; Receptors, Estrogen

Impressive data from the many studies on cultured bone cells and rat models of postmenopausal osteoporosis support a significant bone-sparing effect of the soy isoflavones genistein and daidzein. Translating this research to the clinic has been more challenging, and thus far only a few clinical studies have attempted to tease out the influence of phytoestrogens on bone from the many other components of the diet. Human studies have shown promising although variable results. Studies have been mostly of short duration and with relatively small sample sizes, making it difficult to observe significant and accurate changes in bone. Levels of intake of the soy protein and isoflavones are varied, and the optimal isoflavone intake for bone-sparing effects remains to be determined. Clinical studies thus far performed can be broadly divided into those that have assessed biochemical evidence of reduced bone turnover from measurement of surrogate markers of osteoblast and osteoclast activity, and those that have examined changes in bone mineral density. There are no studies examining effects on fracture rate. This review focuses specifically on the potential influence of phytoestrogens on bone by examining the evidence from 17 in vitro studies of cultured bone cells, 24 in vivo studies of animal models for postmenopausal osteoporosis, 15 human observational/epidemiologic studies, and 17 dietary intervention studies. On balance, the collective data suggest that diets rich in phytoestrogens have bone-sparing effects in the long term, although the magnitude of the effect and the exact mechanism(s) of action are presently elusive or speculative.

Topics: Animals; Biomarkers; Bone and Bones; Clinical Trials as Topic; Disease Models, Animal; Estrogens, Non-Steroidal; Female; Humans; Isoflavones; Osteoporosis, Postmenopausal; Phytoestrogens; Plant Preparations

The use of dietary phyto-oestrogens as a possible option for the prevention of osteoporosis has raised considerable interest because of the increased concern about the risks associated with the use of hormone-replacement therapy. However, the evidence in support of a bone-sparing effect in post-menopausal women is still not sufficiently convincing. Most studies have been performed on soyabean isoflavones (genistein and daidzein), either in the purified form or as a soyabean-based product or extract. In vitro studies using primary cell cultures or stabilised cell lines indicate that treatment with genistein may lead to a reduction in bone resorption, but effects on bone formation have also been shown. Investigations using animal models have provided convincing evidence of major improvements in bone mass or bone turnover following soyabean feeding. Cross-sectional observations in South-East Asian populations with moderately high intakes of soyabean isoflavones (50 mg/d) have shown that women in the high quartile of intake have higher bone mineral density (BMD) and reduced bone turnover, an effect that has not been shown in populations with low average intakes. Human trials have given an indication of a possible effect on lumbar spine BMD, although they have been either short term (<6 months) or methodologically weak. Unresolved issues are: the optimal dose compatible with safety; the individual differences in response that can be related to diet and genotypes; the duration of exposure. Furthermore, there needs to be an evaluation of the relative biological effects of phyto-oestrogens other than isoflavones (lignans, resorcylic acid lactones, flavanols, coumestans) that are also present in European diets.

Topics: Animals; Bone and Bones; Bone Density; Bone Development; Bone Resorption; Disease Models, Animal; Glycine max; Humans; Isoflavones; Osteoporosis; Phytoestrogens; Plant Preparations

Epidemiological data of phytoestrogens and prostate cancer strongly supports the cancer protective effects of isoflavones found in soy products. Inhibition of cell proliferation via hormone-dependent and hormone-independent mechanisms by soy phytochemicals has been studied extensively in cell culture and animal studies. Herein, we review the current literature on the epidemiology and effects of two soy phytoestrogens, genistein and daidzein, and would stress the need for controlled human trials to assess the true preventive and therapeutic effects of these compounds.

Topics: Animals; Disease Models, Animal; Estrogens, Non-Steroidal; Genistein; Glycine max; Humans; Isoflavones; Male; Phytoestrogens; Plant Preparations; Prostate-Specific Antigen; Prostatic Neoplasms; Rats; Rats, Wistar; Signal Transduction

Topics: Animals; Colonic Neoplasms; Disease Models, Animal; Epidemiologic Studies; Estrogens, Non-Steroidal; Glycine max; Humans; Isoflavones; Phytoestrogens; Plant Preparations; Rats; Risk Factors; Soybean Proteins

This article documents and quantitatively assesses the capacity of estrogen, phytoestrogens, and antiestrogens to affect biphasic dose-response relationships in animal/human models and across a broad range of cell types, affecting multiple endpoints. The range of endpoints displaying such biphasic dose responses includes plasminogen activation, oxytocin secretion, angiogenesis, cell proliferation, bone growth, monocyte chemotaxis, secretion of various cytokines, and other effects. The quantitative features of the dose response relationships revealed that the magnitude of the stimulatory responses was typically less than twofold, whereas the stimulatory responses were markedly variable ranging from about 5- to 10(6)-fold. Mechanistic explanations of the biphasic responses are addressed.

Topics: Animals; Bone Development; Cell Division; Chemotaxis; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Estrogen Receptor Modulators; Estrogens; Estrogens, Non-Steroidal; Homeostasis; Humans; Isoflavones; Monocytes; Neovascularization, Physiologic; Oxytocin; Phytoestrogens; Plant Preparations; Plasminogen Activators

The evidence that natural isoflavones protect against several chronic diseases is both observational and experimental. In humans, epidemiologic findings clearly show a higher incidence of some common types of cancer (i.e., breast, prostate, and colon) and of coronary heart diseases in Western populations exposed to limited amounts of soybean isoflavones (i.e., genistein, daidzein) in the diet. Further evidence for cancer and cardiac protection and antiatherogenic effects resulting from soybean isoflavones administration has been noted in various experimental animal models. Isoflavones may also prevent postmenopausal bone loss and osteoporosis. In fact, genistein has been reported to be as active as estrogens in maintaining bone mass in ovariectomized rats. Moreover, the synthetic isoflavone derivative ipriflavone is able to reduce bone loss in various types of animal models of experimental osteoporosis providing a rationale on its use in the prevention and treatment of postmenopausal and senile osteoporosis in humans. The mechanism through which isoflavones may exert the above-mentioned effects seems to depend, at least in part, on their mixed estrogen agonist-antagonist properties. An alternative hypothetical mechanism could derive from other biochemical actions of isoflavones such as inhibition of enzymatic activity, in particular protein kinases, or activation of an "orphan" receptor distinct from the estrogen type I receptor.

Topics: Animals; Coronary Disease; Diet; Disease Models, Animal; Estrogens, Non-Steroidal; Female; Glycine max; Humans; Isoflavones; Neoplasms; Osteoporosis, Postmenopausal; Phytoestrogens; Plant Preparations; Plants; Receptors, Estrogen

Reliable and adequate animal models are required, not only for investigation of etiology, pathogenesis, and treatment of prostate cancer, but also for chemoprevention of prostatic carcinogenesis.. Animal models for the study of premalignant changes in the prostate are reviewed in the paper, with specific reference to the neonatally estrogenized mouse model.. Neonatal treatment of newborn Han:NMRI mice with synthetic non-steroidal estrogen, diethylstilbestrol (DES; 2 micrograms/pup on days 1-3 after birth) promoted hyperplastic and dysplastic changes in the periurethral region of the prostate at the age of 9-18 months. Dietary soy partially inhibited the development of prostatic dysplasia in these neonatally estrogenized animals, which may be due to phytoestrogens contained in soy-rich food.. Prostatic cancer and its possible precursors develop spontaneously, or can be induced by different chemical and hormonal manipulations in certain animal species and strains. Neonatal estrogenization of the mouse results in prostatic dysplasia, which can be partially prevented by dietary soy. There are morphological similarities between human prostatic intraepithelial neoplasia (PIN) and dysplastic changes in rodent prostates, but more data is needed before these dysplastic lesions can be considered equivalent to human PIN.

Topics: Animals; Animals, Newborn; Diet; Diethylstilbestrol; Disease Models, Animal; Estrogens, Non-Steroidal; Glycine max; Isoflavones; Male; Mice; Phytoestrogens; Plant Preparations; Prostate; Prostatic Hyperplasia; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Rats; Rats, Inbred F344

Topics: Animals; Antioxidants; Diet; Disease Models, Animal; Estrogens; Estrogens, Non-Steroidal; Female; Genistein; Growth Inhibitors; Heart Diseases; Humans; Isoflavones; Male; Neoplasms; Neoplasms, Experimental; Neovascularization, Pathologic; Phytoestrogens; Plant Preparations

Microglia, resident immune cells in the brain, are shown to mediate the crosstalk between psychological stress and depression. Interestingly, increasing evidence indicates that sex hormones, particularly estrogen, are involved in the regulation of immune system. In this study, we aimed to understand the potential effects of chronic social defeat stress (CSDS) and genistein (GEN), an estrogenic compound of the plant origin, on neuron-microglia interactions in the mouse hippocampus. The time spent in the avoidance zone in the social interaction test was increased by CSDS 1 day after the exposure, while the avoidance behavior returned to control levels 14 days after the CSDS exposure. Similar results were obtained from the elevated plus-maze test. However, the immobility time in the forced swim test was increased by CSDS 14 days after the exposure, and the depression-related behavior was in part alleviated by GEN. The numerical densities of microglia in the hippocampus were increased by CSDS, and they were decreased by GEN. The voxel densities of synaptic structures and synaptic puncta colocalized with microglia were decreased by CSDS, and they were increased by GEN. Neither CSDS nor GEN affected the gene expressions of major pro-inflammatory cytokines. Conversely, the expression levels of genes related to neurotrophic factors were decreased by CSDS, and they were partially reversed by GEN. These findings show that GEN may in part alleviate stress-related symptoms, and the effects of GEN may be associated with the modulation of neuron-microglia signaling via chemokines and neurotrophic factors in the hippocampus.

Topics: Animals; Behavior, Animal; Depression; Disease Models, Animal; Genistein; Hippocampus; Mice; Microglia; Phytoestrogens; Signal Transduction; Social Defeat; Stress, Psychological; Synapses

The effective treatment of polycystic ovary syndrome (PCOS)-related hormonal disorders necessitates the development of novel treatment strategies. Resveratrol is found in certain food products, and is known to exhibit phytoestrogen properties. The present study was to assess whether resveratrol exhibits beneficial phytoestrogenic effects and associated hormonal modulation in a rat model of PCOS.. This model was established by administering oral letrozole to female Sprague-Dawley (SD) rats prior to randomizing them into control, model and resveratrol treatment groups (40, 80, or 160 mg/kg). Animals were treated for 30 days, after which time ovarian tissues were collected and evaluated. We found that resveratrol administration was associated with increased levels of plasma adiponectin and estradiol levels and restoration of normal ovarian morphology in PCOS model animals. In addition, this treatment was linked to the increased ovarian expression of nesfatin-1 and aromatase at the RNA and protein levels.. Together things findings suggest that resveratrol may represent an effective tool for treating PCOS owing to its phytoestrogenic properties.

Topics: Adiponectin; Animals; Aromatase; Aromatase Inhibitors; Disease Models, Animal; Estradiol; Female; Letrozole; Nucleobindins; Ovary; Phytoestrogens; Polycystic Ovary Syndrome; Random Allocation; Rats; Resveratrol

The antioxidant and anti-inflammatory features of Formononetin, an isoflavone constituent extracted from traditional Chinese medicine, have been reported. The present study investigated that whether Formononetin plays a benefit on hyperoxic ALI.. C57BL/6 mice were exposed to hyperoxia for 72 h to produce experimental hyperoxic ALI model. Formononetin or vehicle was administrated intraperitoneally. Samples from the lung were collected at 72 h post hyperoxia exposure for further study. Pulmonary microvascular endothelial cells isolated from the lung of C57BL/6 mice were used for in vitro study.. Formononetin pretreatment notably attenuated hyperoxia-induced elevating pulmonary water content, upregulation of proinflammatory cytokine levels and increasing infiltration of neutrophil in the lung. Western blot analyses showed that Formononetin enhanced the expression of nuclear factor erythroid-2-related factor 2 (Nrf2) which is a key transcription factor regulating the expression of heme oxygenase-1 (HO-1). Formononetin increased HO-1 expression and activity compared with vehicle-treated animals. Moreover, Formononetin reversed hyperoxia-caused the reduction of M2 macrophage polarization. However, pretreatment of a HO-1 inhibitor reduced the protective effect of Formononetin on hyperoxic ALI. Cell study showed that the Formononetin-induced upregulation of HO-1 was abolished when the Nrf2 was silenced.. Formononetin pretreatment reduces hyperoxia-induced ALI via Nrf2/HO-1-mediated antioxidant and anti-inflammatory effects.

Topics: Acute Lung Injury; Animals; Disease Models, Animal; Endothelial Cells; Heme Oxygenase-1; Hyperoxia; Isoflavones; Lung; Membrane Proteins; Mice; NF-E2-Related Factor 2; Phytoestrogens

The use of animal models in fundamental or pre-clinical research remains an absolute requirement for understanding human pathologies and developing new drugs. In order to transpose these results into clinical practice, many parameters must be taken into account to limit bias. Attention has recently been focused on the sex, age or even strain of each animal, but the impact of diet has been largely neglected. Soy, which is commonly used in the diet in varying quantities can affect their physiology. In order to assess whether the presence of soy can impact the obtained results, we studied the impact of a soy-based diet

Topics: Animal Feed; Animals; Diastole; Diet; Disease Models, Animal; Female; Glycine max; Heart Ventricles; Humans; Male; Phytoestrogens; Rats; Rats, Transgenic; Receptors, Adrenergic, beta-3

Diabetes-induced oxidative stress is vital in initiating neuronal damage in the diabetic retina, leading to diabetic retinopathy (DR). This study investigates the possible effects of coumestrol (CMS) on streptozotocin (STZ)-induced DR. First, we established a rat model of DR by STZ injection and a cell model involving high-glucose (HG) exposure of human retinal microvascular endothelial cells (hRMECs). We characterized the expression patterns of oxidative stress indicators, pro-inflammatory cytokines, and pro-apoptotic proteins in hRMECs. Polymerase chain reaction showed sirtuin 1 (SIRT1) to be poorly expressed in the retinal tissues of STZ-treated rats and HG-exposed hRMECs, but its expression was upregulated upon treatment with CMS treatment. Furthermore, CMS treatment attenuated the STZ-induced pathologies such as oxidative stress, inflammation, and cell apoptosis. Consistent with the

Topics: Animals; Apoptosis; Coumestrol; Diabetes Mellitus, Experimental; Diabetic Retinopathy; Disease Models, Animal; Endothelial Cells; Glucose; Humans; Inflammation; Oxidative Stress; Phytoestrogens; Rats; Retina; Retinal Vessels; Sirtuin 1

Topics: Animals; Cognitive Dysfunction; Cytokines; Disease Models, Animal; Estradiol; Female; Gene Expression Regulation; Hippocampus; Malondialdehyde; Maze Learning; Mice; Ovariectomy; Oxidative Stress; Phytoestrogens; Pueraria

Supplementation with phytoestrogens and insoluble fibers has been reported to reduce duodenal polyps in colectomized familial adenomatous polyposis patients, with a mechanism involving, at least in part, upregulation of estrogen receptor-β subtype, whose expression is lowered during intestinal tumorigenesis. These data suggest a protective effect also in the colon, the main target organ for tumorigenesis in familial adenomatous polyposis and a major cancer type in non-familial (sporadic) cancers. Therefore, we tested whether a similar preparation might reduce tumorigenesis in the colon of Pirc rats (F344/NTac-Apc) mutated in the Apc gene and thus, like familial adenomatous polyposis patients, spontaneously developing multiple tumors in the colon. We first demonstrate that estrogen receptor-β expression in Pirc rat colon is significantly down-regulated compared to age-matched wt rats. Then, Pirc rats aged 1 month were treated for 3 months with Adipol (Adi), a patented preparation containing phytoestrogens and insoluble fibers. Colon tumorigenesis was significantly reduced by Adi treatment (colon tumors/rat were 5.3 ± 0.8 and 2.9 ± 0.3, Mucin Depleted Foci/rat 127 ± 6.6 and 97.1 ± 8.6 in Controls and Adi-treated rats, respectively, means ± SE, P < 0.01). The treatment also normalized colon proliferation pattern along the crypt and significantly increased apoptosis in colon tumors. Estrogen receptor-β expression was increased by Adi treatment, especially in the tumors. These positive effects suggest that Adipol may be exploited as a chemopreventive agent to reduce cancer risk in familial adenomatous polyposis patients and to postpone prophylactic colectomy. Moreover, given the similarities between familial adenomatous polyposis and sporadic colorectal cancer, it might also be used as chemopreventive agent in colorectal cancer patients at risk.

Topics: Adenomatous Polyposis Coli; Adenomatous Polyposis Coli Protein; Animals; Apoptosis; Carcinogenesis; Colon; Colonic Neoplasms; Dietary Fiber; Dietary Supplements; Disease Models, Animal; Estrogen Receptor beta; Gene Expression Regulation, Neoplastic; Humans; Intestinal Mucosa; Male; Mutation; Phytoestrogens; Rats; Rats, Transgenic

The exact mechanisms of polycystic ovary syndrome (PCOS) are unknown and there is no effective cure for the disease. The aim of this study was to evaluate the alterations in serum oestradiol and adiponectin levels and in the expression of some important genes in the uterine and ovarian tissues of PCOS rats. The therapeutic effect of quercetin on PCOS was also assessed. Rats were divided into five groups: control, ethanol, quercetin (Q), PCOS and PCOS+Q. After 30 days of oral treatments, the rats' ovaries and uteri were removed and nesfatin-1, aromatase and adipoR1 expressions were quantified with real-time polymerase chain reaction. Serum adiponectin and oestradiol levels were evaluated using enzyme-linked immunosorbent assay technique. The results of this study showed that expression of nesfatin-1 and adipoR1 genes and adiponectin serum levels decreased in the PCOS rats, but aromatase expression and oestradiol level increased. Treatment with quercetin increased the adiponectin level and expression of adipoR1 and nesfatin-1 and decreased both the expression of aromatase and the oestradiol level. Quercetin improved PCOS by phytoestrogenic effects and mimicking oestrogen's function. Quercetin also affects important factors in both the uterus and ovary and could improve the obesity and the diabetic and infertility symptoms of PCOS.

Topics: Adiponectin; Animals; Aromatase; Dehydroepiandrosterone; Disease Models, Animal; Estradiol; Female; Nucleobindins; Ovary; Phytoestrogens; Polycystic Ovary Syndrome; Quercetin; Rats, Sprague-Dawley; Receptors, Adiponectin; Uterus

Estrogenic deficiency is considered a risk of coronary disease in women. The phytoestrogen genistein could be a safe preventive strategy. The first aim of this work was to validate a model of cardiac stunning in which natural estrogenic deficiency rats, ie, adult young male (YM) and aged female (AgF), are compared with young female rats (YF). The second aim was to study whether the in vivo administration of genistein prevents the stunning in estrogenic deficiency rats. The third aim was to evaluate whether in our estrogenic deficiency model exists a synergy between genistein and estradiol. The fourth aim was to characterize the underlying mechanisms of genistein. Stunning was induced by ischemia/reperfusion (I/R) in isolated hearts inside a calorimeter. The left ventricular pressure (P) and total heat rate (Ht) were simultaneously measured, while diastolic contracture and muscle economy (P/Ht) were calculated. During R, P/Ht and P recovered less in AgF and YM than in YF rat hearts. Genistein through i.p. (GST-ip) improved P and P/Ht in AgF and YM, but not in YF. In YM, the cardioprotections of GST-ip and estradiol were synergistic. After ischemia, GST-ip increased SR Ca leak causing diastolic contracture. The GST-ip cardioprotection neither was affected by blockade of PI3K-Akt, NO synthases, or phosphatases, but it was sensitive to blockade of protein-kinase C and mKATP channels. Results suggest that (1) estrogenic deficiency worsens cardiac stunning, (2) GST-ip was more cardioprotective in estrogenic deficiency and synergistic with estradiol, and (3) cardioprotection of GST-ip depends on the protein-kinase C and mKATP channel pathway activation.

Topics: Age Factors; Animals; Calcium Signaling; Disease Models, Animal; Energy Metabolism; Estradiol; Female; Genistein; Isolated Heart Preparation; Male; Mitochondria, Heart; Myocardial Reperfusion Injury; Myocardial Stunning; Myocytes, Cardiac; Phytoestrogens; Potassium Channels; Protein Kinase C; Rats, Sprague-Dawley; Sex Factors; Ventricular Function, Left; Ventricular Pressure

Trigonelline is a potent phytochemical present in fenugreek, which has strong anti-oxidant and phytoestrogenic activities. This study was carried out to investigate this estrogenic activity as a possible mechanisms involved in preventing the symptoms of osteoporosis in dexamethasone induced osteoporosis in rats.. Wistar rats were randomly divided into eight groups, six animals in each group. Osteoporosis was induced using dexamethasone 0.1mg/kg subcutaneously in rats for three times per week for 8 consecutive weeks and treatment with drugs up to 12 weeks as per the treatment schedule described. After 12 weeks, rats were sacrificed; blood samples were collected from each rat and the clear, non hemolysed supernatant sera was used for biochemical examinations. Femurs were used for Bone Mineral Density (BMD), microcomputed tomography (Micro CT), histology and biochemical examinations.. BMD, bone micro structure, serum calcium, phosphorus level and serum estradiol levels were decreased while serum PTH levels, SAP and acid phosphatase (ACP) were elevated in dexamethasone treated rats as compared to control (p<0.01). Dexmethasone treated animals showed loss of marrow at multifocal area, cartilage and trabeculae and thinning of trabeculae (bone resorption), zone of cartilage was poorly seen and fat cells in marrow. Trigonelline showed significant improvement and prevent the progression of osteoporosis by enhancing the BMD, restoring bone physiology.. Our results confirm the estrogenic activity of triogonelline, which is responsible for its effects; still, it needs further evaluation in other animal models to provide a more conclusive view for its therapeutic usefulness in osteoporosis.

Topics: Alkaloids; Animals; Bone Density; Dexamethasone; Disease Models, Animal; Femur; Humans; Osteoporosis; Phytoestrogens; Rats; Rats, Wistar; Trigonella; X-Ray Microtomography

Estrogen is involved in lipid metabolism. Menopausal women with low estrogen secretion usually gain weight and develop steatosis associated with abnormal lipid metabolism. A previous study showed that blackcurrant (

Topics: Adipocytes; Adiponectin; Alanine Transaminase; Animals; Aspartate Aminotransferases; Body Weight; Cholesterol, HDL; Cholesterol, LDL; Diet, High-Fat; Dietary Supplements; Disease Models, Animal; Dyslipidemias; Fatty Liver; Female; gamma-Glutamyltransferase; Intra-Abdominal Fat; Leptin; Lipid Metabolism; Liver; Menopause; Non-alcoholic Fatty Liver Disease; Ovariectomy; Phytoestrogens; Plant Extracts; Rats; Rats, Sprague-Dawley; Ribes; Triglycerides

Chronic neurocognitive impairments, commonly associated with pediatric human immunodeficiency virus type 1 (PHIV), are a detrimental consequence of early exposure to HIV-1 viral proteins. Strong evidence supports S-Equol (SE) as an efficacious adjunctive neuroprotective and/or neurorestorative therapeutic for neurocognitive impairments in adult ovariectomized female HIV-1 transgenic (Tg) rats. There remains, however, a critical need to assess the therapeutic efficacy of SE when treatment occurs at an earlier age (i.e., resembling a therapeutic for children with PHIV) and across the factor of biological sex. Utilization of a series of signal detection operant tasks revealed prominent, sex-dependent neurocognitive deficits in the HIV-1 Tg rat, characterized by alterations in stimulus-reinforcement learning, the response profile, and temporal processing. Early (i.e., postnatal day 28) initiation of SE treatment precluded the development of chronic neurocognitive impairments in all (i.e., 100%) HIV-1 Tg animals, albeit not for all neurocognitive domains. Most notably, the therapeutic effects of SE are generalized across the factor of biological sex, despite the presence of endogenous hormones. Results support, therefore, the efficacy of SE as a neuroprotective therapeutic for chronic neurocognitive impairments in the post-cART era; an adjunctive therapeutic that demonstrates high efficacy in both males and females. Optimizing treatment conditions by evaluating multiple factors (i.e., age, neurocognitive domains, and biological sex) associated with PHIV and HIV-1 associated neurocognitive disorders (HAND) affords a key opportunity to improve the therapeutic efficacy of SE.

Topics: Animals; Child; Cognitive Dysfunction; Conditioning, Operant; Disease Models, Animal; Female; HIV Infections; HIV-1; Humans; Male; Mental Status and Dementia Tests; Neuroprotective Agents; Phytoestrogens; Rats; Rats, Inbred F344; Rats, Transgenic; Reinforcement, Psychology; Sex Factors

Phytoestrogens are nonsteroidal plant compounds with similar chemical structures to mammalian estrogen capable of mimicking the effect of estrogen in selective tissues. A diet rich in phytoestrogens is associated with a variety of health benefits including decreased risks for heart disease, breast cancer, and osteoporosis. Obesity has long thought to be associated with improved bone density due to increased mechanical loading, but recent literature suggests obesity may actually decrease bone health. Daidzein, a soy-derived phytoestrogen, has been shown to improve parameters of bone health in lean animal models of osteoporosis but has not been tested in obese animals. Following a one-week acclimation to a standard AIN-93G diet, 19 five-week-old female obese Zucker rats (OZR) were randomly assigned to a modified AIN-93G diet containing either high daidzein (HD, 0.121 g kg-1 feed) or low daidzein (LD, 0.01 g kg-1 feed). After 8 weeks, tibias and femurs were removed to assess true density (Archimedes principal), mechanical strength (three-point bending test), and femoral osteogenic gene expression. Serum was collected to assess osteocalcin and deoxypyridinoline. Our results indicated that there were no significant differences between the measures for tibial or femoral true density or mechanical strength for the rats in the HD and LD diet groups. Similarly, there were no significant differences in gene expressions related to osteogenic pathways, or serum biomarkers of bone formation and resorption. Overall, an increased dose of daidzein from soy protein supplementation does not elicit an improvement in markers of bone health in obese Zucker rats.

Topics: Amino Acids; Animals; Biomarkers; Body Weight; Bone and Bones; Bone Density; Diet; Dietary Supplements; Disease Models, Animal; Energy Intake; Female; Femur; Gene Expression; Isoflavones; Obesity; Osteocalcin; Osteogenesis; Osteoporosis; Phytoestrogens; Rats; Rats, Zucker

One of the major female sex hormones, estrogen, can influence a variety of mental states. Individuals with multiple sclerosis (MS) often suffer from mental health issues, which are correlated with the pathology of gray matter. In this study, we aimed to elucidate the validity of phytoestrogen genistein (GEN) for treating the gray matter lesions in MS using the mouse model of cuprizone (CPZ)-induced demyelination. First, we confirmed that 5-week 0.2% CPZ intoxication induced demyelination in the hippocampus, and that myelination was successfully recovered by GEN. Loss of mature oligodendrocytes following CPZ intoxication was counteracted by GEN. Neither CPZ nor GEN affected the densities of oligodendrocyte precursor cells and astrocytes. CPZ-induced activation and proliferation of microglia were not inhibited by GEN. Upregulation of gene expression of the pro-inflammatory cytokine, interleukin-1β, in sorted microglia by CPZ was not inhibited by GEN either. However, the expression levels of genes related to phagocytosis, such as cluster of differentiation 68 and lysosomal-associated membrane protein 1, in sorted microglia were elevated by CPZ but lowered by GEN. Notably, physical contact of microglia with myelin was increased by CPZ but decreased by GEN. The expression levels of myelin-related genes, such as myelin basic protein and myelin oligodendrocyte glycoprotein, in the whole hippocampal tissue were decreased by CPZ but recovered by GEN. These results show that GEN may act on mature oligodendrocytes in the hippocampus by promoting their survival and myelin formation, and suggest the therapeutic potential of phytoestrogens for treating MS patients suffering from mental health issues.

Topics: Animals; Cell Differentiation; Cell Survival; Cuprizone; Disease Models, Animal; Down-Regulation; Female; Genistein; Hippocampus; Humans; Male; Mice; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Myelin Sheath; Phagocytosis; Phytoestrogens; Treatment Outcome

Oxidant toxicity has been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS), an insidiously progressive neurodegenerative disorder involving upper and lower motor neurons. Here, we investigated the cellular and molecular mechanisms underlying the neuroprotective effects of an anti-oxidant genistein in SOD1-G93A transgenic mouse model of ALS. Rotarod test, hanging wire test and hindlimb clasping test were used to determined disease onset and assess motor performance. Immunostaining together with neuronal size measurement were used to count viable motor neurons. In addition, immunostaining procedure and ELISA kit were used to assess the inflammatory response in the spinal cord. Our results showed that Genistein administration suppressed the production of pro-inflammatory cytokines and alleviated gliosis in the spinal cord of SOD1-G93A mice. In addition, genistein administration induced autophagic processes and enhanced the viability of spinal motor neurons. As a result, genistein alleviated ALS-related symptoms and slightly prolonged the lifespan of SOD1-G93A mice. Taken together, our results indicate that genistein is neuroprotective in SOD1-G93A mice, suggesting genistein could be a promising treatment for human ALS. Graphical Abstract Genistein protects impariments in SOD1-G93A transgenic mouse model.

Topics: Amyotrophic Lateral Sclerosis; Animals; Disease Models, Animal; Female; Genistein; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neuroprotective Agents; Phytoestrogens; Superoxide Dismutase

Postmenopausal osteoporosis is a common disorder accompanied with estrogen deficiency in women. Plants containing phytoestrogens and amino acids have been used in the osteoporosis treatment. The present study aims to evaluate the estrogen-like activity of the Cicer arietinum extract (CAE) and its ability to inhibit osteoclastogenesis process. These achieved by investigating the binding of its active phytoestrogens (genistein, daidzein, formononetin and biochanin A) to the estrogen receptors (ER) α and β of rats and human in silico. In addition, in vivo study on ovariectomized (OVX) rats is performed. For in vivo study, twenty four rats were divided into four groups (n= 6). Group I is the sham control rats which administered distilled water. Groups II, III, and IV are OVX groups which administered distilled water, CAE (500 mg/kg), and alendronate; respectively. The docking study revealed that the phytoestrogens docked into the protein active site with binding energies comparable with that of estrogens (estriol and β-estradiol) which means the similarity between the estrogenic contents of CAE and the ensogenous ones. Additionally, in vivo study revealed that CAE reverse TRAP5b and RANKL levels that drastically increased in the untreated OVX group. But, it trigger upregulation of OPG, enhance the OPG/RANKL ratio and modulate the bone and uterus alterations of OVX group. Phytoestrogens and the bone-protective amino acids contents of CAE could be responsible for their estrogen-like effect and antiosteoporotic activity. These results concluded that CAE is an attractive candidate for developing a potential therapeutic cheap agent used as an alternative to the synthetic estrogen replacement therapy. Further, in vivo validation is required for its clinical application.

Topics: Alendronate; Animals; Bone Density Conservation Agents; Calcium-Binding Proteins; Cicer; Disease Models, Animal; Estradiol; Estrogen Receptor alpha; Estrogen Receptor beta; Female; Gene Expression Regulation; Genistein; Humans; Isoflavones; Membrane Glycoproteins; Molecular Docking Simulation; Osteogenesis; Osteoporosis; Osteoprotegerin; Ovariectomy; Phytoestrogens; Phytotherapy; Protein Structure, Secondary; RANK Ligand; Rats; Receptors, Cytoplasmic and Nuclear; Receptors, Peptide

Radix Astragali, has long been used to alleviate allergic diseases (ADs). Formononetin is one of the major active components in Radix Astragali, but its mechanism on ADs is not definitively known. The fluorescein isothiocyanate isomer-induced atopic contact dermatitis mouse model and poly I:C or lipopolysaccharide-treated HaCaT cells were used to examine thymic stromal lymphopoietin (TSLP)/interleukin (IL)-33 production and expression of E-cadherin. After administration of formononetin, TSLP/IL-33 levels decreased both in vitro and in vivo, while E-cadherin was increased in vivo and restored in vitro. Furthermore, small interference RNA silencing of E-cadherin resulted in elevated levels of TSLP, whereas the inhibitory effect of formononetin on TSLP was no longer observed. In addition, TSLP resulted in no detectable changes in delocalization or protein expression of E-cadherin in HaCaT cells. These results indicated that formononetin showed a protective effect in ADs, which was correlated with decreasing TSLP/IL-33 production via regulation of E-cadherin.

Topics: Animals; Astragalus propinquus; Cadherins; Cells, Cultured; Cytokines; Dermatitis, Allergic Contact; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Humans; Interleukin-33; Isoflavones; Mice; Mice, Inbred BALB C; Phytoestrogens; RNA, Small Interfering; Thymic Stromal Lymphopoietin

The bone regeneration and healing effect of formononetin was evaluated in a cortical bone defect model that predominantly heals by intramembranous ossification. For this study, female Balb/c mice were ovariectomised (OVx) and a drill-hole injury was generated in the midfemoral bones of all animals. Treatment with formononetin commenced the day after and continued for 21 d. Parathyroid hormone (PTH1-34) was used as a reference standard. Animals were killed at days 10 and 21. Femur bones were collected at the injury site for histomorphometry studies using microcomputed tomography (μCT) and confocal microscopy. RNA and protein were harvested from the region surrounding the drill-hole injury. For immunohistochemistry, 5 µm sections of decalcified femur bone adjoining the drill-hole site were cut. μCT analysis showed that formononetin promoted bone healing at days 10 and 21 and the healing effect observed was significantly better than in Ovx mice and equal to PTH treatment in many aspects. Formononetin also significantly enhanced bone regeneration as assessed by calcein-labelling studies. In addition, formononetin enhanced the expression of osteogenic markers at the injury site in a manner similar to PTH. Formononetin treatment also led to predominant runt-related transcription factor 2 and osteocalcin localisation at the injury site. These results support the potential of formononetin to be a bone-healing agent and are suggestive of its promising role in the fracture-repair process.

Topics: Animals; Bone Regeneration; Core Binding Factor Alpha 1 Subunit; Cortical Bone; Disease Models, Animal; Fabaceae; Femur; Fractures, Bone; Isoflavones; Mice, Inbred BALB C; Osteocalcin; Osteogenesis; Ovariectomy; Parathyroid Hormone; Phytoestrogens; Phytotherapy; Plant Extracts; Wound Healing

Aucubin (AU) is an iridoid glycoside that has been shown to display estrogenic properties and has various pharmacological effects. Herein, we described the angiogenic properties of AU. In the study, hindlimb ischemia was induced by ligation of femoral artery on the right leg of ovariectomized mice. AU treatment significantly accelerated perfusion recovery and reduced tissue injury in mice muscle. Quantification of CD31-positive vessels in hindlimb muscles provided evidences that AU promoted angiogenesis in peripheral ischemia. In addition, results from quantitative PCR and western blot suggested AU induced angiogenesis via vascular endothelial cell growth factor (VEGF)/Akt/endothelial nitric oxide synthase (eNOS) signaling pathway. More interestingly, AU's angiogenic effects could be completely abolished in estrogen receptor beta (ERβ) knockout mice. In conclusion, the underlying mechanisms were elucidated that AU produced pro-angiogenic effects through ERβ-mediated VEGF signaling pathways. These results expand knowledge about the beneficial effects of AU in angiogenesis and blood flow recovery. It might provide insight into the ERβ regulating neovascularisation in hindlimb ischemia and identify AU as a potent new compound used for the treatment of peripheral vascular disease.

Topics: Angiogenesis Inducing Agents; Animals; Disease Models, Animal; Estrogen Receptor beta; Female; Femoral Artery; Gene Expression Regulation; Hindlimb; Iridoid Glucosides; Ischemia; Ligation; Mice; Mice, Inbred C57BL; Mice, Knockout; Neovascularization, Physiologic; Nitric Oxide Synthase Type III; Ovariectomy; Phytoestrogens; Platelet Endothelial Cell Adhesion Molecule-1; Proto-Oncogene Proteins c-akt; Recovery of Function; Signal Transduction; Vascular Endothelial Growth Factor A

As a phytoestrogen, 3,3'-diindolylmethane (DIM) plays a chemopreventive role by inhibiting cancer progression. In this study, we examined the effects of 17β-estradiol (E2), two endocrine disrupting chemicals (EDCs), triclosan (TCS) and bisphenol A (BPA), and DIM on epithelial-mesenchymal transition (EMT) and metastatic behaviors of estrogen receptor (ER)-positive MCF-7 breast cancer cells. An in vitro assay revealed that E2 (10

Topics: Animals; Benzhydryl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Disease Models, Animal; Endocrine Disruptors; Female; Heterografts; Humans; Indoles; Mice; Mice, Inbred BALB C; Phenols; Phytoestrogens; Receptors, CXCR4; Signal Transduction

Phytoestrogen genistein may be useful to treat pulmonary arterial hypertension (PAH). However, its mechanism is still not clear. The aim of the present study was to confirm the therapeutic effects of phytoestrogen genistein on PAH in monocrotaline-induced rat model and to explore its mechanism.. Sprague-Dawley male rats were randomly divided into 4 groups: control group (n=8), PAH group (n=8), genistein treament group with three different doses (n=8 in each dose group) and group of PI3K inhibitor LY294002. The rat model of PAH was induced by monocrotaline (MCT). The situation of survival of rats was observed. Pathological studies of lung and heart tissues were performed. Western-blot detection of P-Akt and P-eNOS expression levels in lung tissue was carried out. Nitrate reductase analysis was used to measure nitric oxide (NO) in lung tissue.. Genistein treatment resulted in significant improvement in the speed of tricuspid regurgitation, diameter of pulmonary artery, mean pulmonary artery pressure and right ventricular hypertrophy index. Genistein treatment also resulted in significant improvement in the stenosis of pulmonary artery, proliferation of smooth muscle, right ventricular hypertrophy and myocardial hypertrophy. These therapeutic effects were more obvious with increasing dose of genistein. After genistein treatment, amelioration in survival rates of PAH rats was observed. PI3K inhibitor LY294002 could block these therapeutic effects. In rat lung tissue, P-Akt, P-eNOS and NO expressions were increased significantly in genistein treatment group when compared with PAH group (p<0.05, respectively). The increase in expression level of P-Akt, P-eNOS and NO was correlated with genistein dose. P-Akt, P-eNOS and NO expressions in lung tissue increased slightly in the PI3K inhibitor LY294002 group when compared with PAH group, but the difference was not statistically significant (p>0.05).. We confirmed that genistein could relax pulmonary vascular resistance, reduce pulmonary artery pressure, improve right heart function and ameliorate survival rate in the rat model of PAH. Our study suggested that its mechanism was related with PI3K/Akt/eNOS signal pathway. Phytoestrogen genistein may become a new and effective drug for patients with PAH.

Topics: Animals; Blotting, Western; Disease Models, Animal; Genistein; Heart Ventricles; Hypertension, Pulmonary; Lung; Male; Monocrotaline; Nitric Oxide Synthase Type III; Phosphatidylinositol 3-Kinases; Phytoestrogens; Proto-Oncogene Proteins c-akt; Random Allocation; Rats; Rats, Sprague-Dawley; Signal Transduction

Soy is the main source of phytoestrogens, which has long been used as traditional food. One major subtype of phytoestrogens includes isoflavones and they are scientifically validated for their beneficial actions on many hormone-dependent conditions.. The present study examines the effect of soy isoflavones on letrozole-induced polycystic ovary syndrome (PCOS) rat model.. PCOS was induced in Sprague-Dawley rats with of 1 mg/kg letrozole, p.o. once daily for 21 consecutive days. Soy isoflavones (50 and 100 mg/kg) was administered for 14 days after PCOS induction. Physical parameters (body weight, oestrous cycle determination, ovary and uterus weight) metabolic parameters (oral glucose tolerance test, total cholesterol), steroidal hormone profile (testosterone and 17β-oestradiol), steroidogenic enzymes (3β-hydroxy steroid dehydrogenase (HSD) and 17β-HSD), oxidative stress and histopathology of ovary were studied.. Soy isoflavones (100 mg/kg) treatment significantly altered the letrozole-induced PCOS symptoms as observed by decreased body weight gain (p < 0.05), percentage diestrous phase (p < 0.001), testosterone (p < 0.001), 3β-HSD (p < 0.01) and 17β-HSD (p < 0.001) enzyme activity and oxidative stress. Histological results reveal that soy isoflavones treatment in PCOS rats resulted in well-developed antral follicles and normal granulosa cell layer in rat ovary.. Treatment with soy isoflavones exerts beneficial effects in PCOS rats (with decreased aromatase activity) which might be due to their ability to decrease testosterone concentration in the peripheral blood.. Analysis of physical, biochemical and histological evidences shows that soy isoflavones may be beneficial in PCOS.

Topics: 17-Hydroxysteroid Dehydrogenases; 3-Hydroxysteroid Dehydrogenases; Androgen Antagonists; Animals; Antioxidants; Biomarkers; Blood Glucose; Cholesterol; Disease Models, Animal; Dose-Response Relationship, Drug; Estradiol; Estrous Cycle; Female; Glycine max; Isoflavones; Letrozole; Nitriles; Ovary; Oxidative Stress; Phytoestrogens; Phytotherapy; Plants, Medicinal; Polycystic Ovary Syndrome; Rats, Sprague-Dawley; Testosterone; Time Factors; Triazoles; Uterus; Weight Gain

Ischemic heart disease is the leading cause of death in postmenopausal women. The expression of caveolin, a membrane protein and a negative regulator of nitric oxide (NO), increases after menopause. The present study was designed to determine the effect of daidzein (DDZ), a phytoestrogen in attenuated cardioprotective effect of ischemic preconditioning (IPC) in ovariectomized rat heart.. Heart was isolated from ovariectomized rat and mounted on Langendorff's apparatus, subjected to 30 min ischemia and 120 min reperfusion. IPC was mediated by four cycles of 5 min ischemia and 5 min reperfusion. The infarct size was estimated using triphenyltetrazolium chloride stain, and coronary effluent was analyzed for lactate dehydrogenase and creatine kinase MB (CK-MB) release to assess the degree of myocardial injury. The release of NO was estimated indirectly by measuring the release of nitrite in coronary effluent.. IPC-induced cardioprotection was significantly attenuated in ovariectomized rats as compared to normal rats, which was restored by treatment of DDZ, a caveolin inhibitor (0.2 mg/kg subcutaneously) for 1 week. However, this observed cardioprotection was significantly attenuated by perfusion of l-nitroarginine methyl ester, an endothelial nitric oxide synthase (eNOS) inhibitor (100 µM/L) and glibenclamide, an adenosine triphosphate-sensitive potassium ion channel blocker (10 µM/L) alone or in combination, noted in terms of increase in myocardial infarct size, release of LDH and CK-MB, and also decrease in the release of NO.. Thus, it is suggested that DDZ restores the attenuated cardioprotective effect in ovariectomized rat heart, which may be due to downregulation of caveolin and subsequent increase in the activity of eNOS.

Topics: Animals; Caveolin 1; Creatine Kinase, MB Form; Disease Models, Animal; Female; Heart; In Vitro Techniques; Ischemic Preconditioning, Myocardial; Isoflavones; L-Lactate Dehydrogenase; Myocardial Infarction; Nitrites; Ovariectomy; Phytoestrogens; Rats, Wistar

Hormone replacement therapy has been used as an effective treatment for the prevention of bone loss in postmenopausal women. In our previous study, QiBaoMeiRan formula (QBMR) had estrogenic activity and could relieve symptoms of hot flushes and body weight increase induced by estrogen decline. However, no evidence base links QBMR to preventing bone loss. The aim of this study is to investigate the effect of QBMR on bone loss.. The ovariectomized rat model was established, and ovariectomized rats were treated with QBMR at doses of 0.875, 1.75, and 3.5 g/kg per day for 8 weeks. Biochemical parameters, bone mineral density, structural morphometric traits and histological characteristics of trabecular bone were assessed.. QBMR treatment significantly decreased the increase in serum alkaline phosphatase, bone Gla-protein and C-telopeptide fragments of type I collagen and decreased the decline of serum calcium and phosphorus in the circulation of ovariectomized rats. QBMR completely corrected the decrease in bone mineral density in lumbar vertebrae (L4-L6) comparable to the sham group. In addition, QBMR treatment also significantly ameliorated the decrease of structural parameters of femur trabecular bone, bone volume fraction, trabecular number, trabecular thickness and bone mineral density as well as the increase in trabecular separation by micro-computerized tomography scanning. These were also confirmed by bone histological results that showed its protective action.. Our results indicated that QBMR had a definite anti-bone loss effect and will have potential to be used for the treatment of postmenopausal osteoporosis.

Topics: Alkaline Phosphatase; Animals; Bone Density; Bone Diseases, Metabolic; Calcium; Collagen Type I; Disease Models, Animal; Drugs, Chinese Herbal; Female; Femur; Lumbar Vertebrae; Ovariectomy; Peptides; Phosphorus; Phytoestrogens; Rats; Rats, Sprague-Dawley

Soy isoflavones have been shown to be an alternative to hormone therapy at menopause, without causing side-effects such as breast cancer. However, the effects of early and late treatment with isoflavones on the mammary gland remain controversial.. To investigate the effects of early and late treatment with soy isoflavones on the mammary gland of ovariectomized rats.. Thirty 3-month-old rats were ovariectomized and divided equally into groups: Control, treated with vehicle solution; or with 150 mg/kg/body weight of isoflavones by gavage; or subcutaneously treated with 10 μg/kg/body weight with 17β-estradiol. Treatments started 3 days (early treatment) or 30 days (late treatment) after ovariectomy and lasted for 30 consecutive days. Thereafter, the animals were euthanized and the mammary glands were removed and processed for paraffin embedding. Sections were stained with hematoxylin and eosin for histomorphometry or subjected to immunohistochemical detection of Ki-67 and VEGF-A.. The ductal, lobular and total epithelial fractions were similar between controls and the early/late isoflavone groups, but they were significantly higher in the groups treated with estradiol. In both epithelial and stromal regions, the immunoreactivity of VEGF-A and the percentage of Ki-67-positive cells were significantly higher in the groups treated with estradiol, while they were similar in the early/late isoflavone groups and control groups.. Our results indicate that early and late treatment with soy isoflavones at the dose of 150 mg/kg/body weight does not show proliferative and angiogenic effects on the mammary gland of ovariectomized rats.

Topics: Animals; Disease Models, Animal; Estradiol; Female; Glycine max; Isoflavones; Ki-67 Antigen; Mammary Glands, Animal; Menopause; Ovariectomy; Phytoestrogens; Random Allocation; Rats; Rats, Wistar; Vascular Endothelial Growth Factor A

Huntington's disease (HD) is a neurodegenerative disorder, characterized by selective atrophy in the striatum, particularly the medium spiny GABAergic efferent neurons. This results in striatal sensorimotor gating deficits. Systemic administration of 3-nitropropionic acid (3-NPA) produces selective lesions mimicking those of HD. Males were found to be more susceptible to 3-NPA-induced neurotoxicity than females, suggesting neuroprotective effects of estrogens. Phytoestrogens, including genistein, are good estrogenic alternatives that keep their beneficial effects on non-reproductive organs and lack the potential hazardous side effects. The current study was designed to investigate the potential beneficial effects of genistein in 3-NPA-induced HD in ovariectomized rats. Results showed that 3-NPA (20 mg/kg) administration caused significant disruption of the rats' locomotor activity and prepulse inhibition. In addition, it decreased striatal ATP levels and increased oxidative stress, inflammatory and apoptotic markers with striatal focal hemorrhage and gliosis. Pretreatment with 17β-estradiol (2.5 mg/kg) or genistein (20 mg/kg) led to a significant improvement of behavioral parameters, increased ATP production, decreased oxidative stress, attenuated inflammation and apoptosis. Therefore, this study suggests potential neuroprotective effects of genistein in ovariectomized rats challenged with 3-NPA.

Topics: Adenosine Triphosphate; Animals; Antioxidants; Apoptosis; Corpus Striatum; Dinoprostone; Disease Models, Animal; Estradiol; Estrogens; Female; Genistein; Huntington Disease; Inflammation Mediators; Locomotion; Neuroprotective Agents; Nitro Compounds; Oxidative Stress; Phytoestrogens; Prepulse Inhibition; Propionates; Rats; Rats, Sprague-Dawley

Autism spectrum disorders (ASDs) are a group of devastating neurodevelopmental syndromes that affect up to 1 in 68 children. Despite advances in the identification of ASD risk genes, the mechanisms underlying ASDs remain unknown. Homozygous loss-of-function mutations in Contactin Associated Protein-like 2 (CNTNAP2) are strongly linked to ASDs. Here we investigate the function of Cntnap2 and undertake pharmacological screens to identify phenotypic suppressors. We find that zebrafish cntnap2 mutants display GABAergic deficits, particularly in the forebrain, and sensitivity to drug-induced seizures. High-throughput behavioral profiling identifies nighttime hyperactivity in cntnap2 mutants, while pharmacological testing reveals dysregulation of GABAergic and glutamatergic systems. Finally, we find that estrogen receptor agonists elicit a behavioral fingerprint anti-correlative to that of cntnap2 mutants and show that the phytoestrogen biochanin A specifically reverses the mutant behavioral phenotype. These results identify estrogenic compounds as phenotypic suppressors and illuminate novel pharmacological pathways with relevance to autism.

Topics: Animals; Animals, Genetically Modified; Autistic Disorder; Disease Models, Animal; Estrogens; Gene Expression Regulation; Genistein; Green Fluorescent Proteins; Humans; Larva; Luminescent Proteins; Membrane Proteins; Motor Activity; Mutation; Nerve Tissue Proteins; Phenotype; Phytoestrogens; Psychotropic Drugs; Seizures; Sleep-Wake Transition Disorders; Vesicular Glutamate Transport Protein 2; Zebrafish

Recent studies have shown that immune system plays a major role in pathophysiology of postmenopausal osteoporosis. Previously we have shown that phytoestrogens like daidzein and medicarpin exhibit immunoprotective effects, by virtue of which they alleviate bone loss. With this background, methoxyisoflavones like formononetin (formo) and isoformononetin (isoformo) that have been studied for preventing bone loss in ovariectomized rats were tested for their immunomodulatory effects in estrogen-deficient bone loss mice model.. Adult Balb/c mice (N = 8/group) were given oral dose of formo and isoformo at 10 mg/kg body weight, post ovariectomy (Ovx) daily for 6 weeks. Animals were autopsied and long bones were harvested to study bone microarchitecture. Peripheral blood mononuclear cells were isolated for fluorescence-activated cell sorting and RNA analysis. Serum was collected for enzyme-linked immunosorbent assay.. It was observed that formo and isoformo treatment to Ovx mice led to significant restoration of Ovx-induced deterioration of trabecular microarchitecture. Pro-osteoclastogenic subset Th17 and B cells were decreased in formo/isoformo-treated Ovx mice in comparison with vehicle-treated Ovx group. Formo and isoformo treatment to Ovx mice also led to decreased expression of Th17 diffentiation factors and promoted T-regulatory cell differentiation. Formo was more effective in enhancing the FOXP3 expression compared with isoformo. IL-17A-induced osteoclastogenesis and inhibition of osteoblast apoptosis were also suppressed by formo and isoformo treatment, with formo having a more potent effect.. Our study demonstrates the immunomodulatory activity of methoxyisoflavones, formo, and isoformo, which translate into improved skeletal parameters, thereby preventing Ovx-induced bone loss.

Topics: Animals; Apoptosis; B-Lymphocytes; Cell Differentiation; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Humans; Isoflavones; Leukocytes, Mononuclear; Lymphopoiesis; Menopause; Mice; Mice, Inbred BALB C; Osteoblasts; Osteogenesis; Osteoporosis, Postmenopausal; Ovariectomy; Phytoestrogens; Th17 Cells

Daidzein is an isoflavone derived from soybeans that exerts preventive effects on bone loss in ovariectomized (OVX) animals. These effects have been correlated with increasing serum equol levels. In the present study, we investigated the effects of antibiotic intake on equol metabolism from daidzein, and the corresponding levels of bone loss in OVX mice.. Eight-week-old female ddY mice (n = 42) were either ovariectomized (OVX) or subjected to a sham operation (sham). OVX mice were then divided into six dietary subgroups: control diet (control), 0.3 % kanamycin diet (KN), 0.1 % daidzein diet (Dz), 0.1 % daidzein and 0.0375 % kanamycin diet (Dz+KN3.75), 0.1 % daidzein and 0.075 % kanamycin diet (Dz+KN7.5), and 0.1 % daidzein and 0.3 % kanamycin diet (Dz+KN30). The mice were fed their respective diets for 4 weeks.. Uterine weight and femoral bone mineral density (BMD) were significantly lower in the OVX mice compared in the sham mice. No significant differences in uterine weight were observed among all OVX dietary subgroups. The Dz subgroup was found to exhibit higher plasma equol and O-desmethylangolensin (O-DMA) concentrations, as well as greater femoral BMD, compared to all other OVX subgroups. Furthermore, when compared to the Dz group, kanamycin intake decreased plasma equol and O-DMA concentrations, as well as femoral BMD in the OVX mice.. These results suggest that kanamycin intake inhibited the conversion of daidzein to equol and O-DMA, blocking the preventive effects of daidzein on bone loss in OVX mice. Therefore, the bone-protective effects of daidzein intake may be predominantly associated with increased plasma concentrations of either equol or O-DMA.

Topics: Administration, Oral; Animals; Biotransformation; Body Weight; Bone Density; Diet; Disease Models, Animal; Equol; Female; Femur; Humans; Isoflavones; Kanamycin; Mice; Organ Size; Osteoporosis; Ovariectomy; Phytoestrogens; Uterus

Oxidative stress is the major cause of neuronal cell degeneration observed in neurodegenerative diseases including vascular dementia (VaD), and hypertension has been found to increase the probability of VaD. Here, we investigated the effects of equol in deoxycorticosterone acetate (DOCA)-salt-induced hypertensive rats (DHRs) and the associated VaD. The systolic blood pressure of rats treated with low- (10 mg per kg body weight) and high-dose (20 mg per kg body weight) equol for 4 weeks was lower than that of the control group by 12.18 and 17.48% in a dose-dependent manner, respectively (p < 0.05), which was regulated by inhibiting angiotensin-converting enzyme (ACE) activity and increasing the nitric oxide (NO) production. Equol-treated DHRs showed a significant decrease in both the swimming distance and time required to reach the escape platform (78.20 to 82.56%, p < 0.05). In addition, the probe trial session and working memory test indicated that equol improved the long- and short-term memory of the rats. Moreover, the brain antioxidant activity was increased by elevating the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels, and the malondialdehyde (MDA) content and acetylcholinesterase (AChE) activity were decreased, indicating that equol suppressed oxidative stress. In conclusion, we demonstrated that equol exhibited comparable blood pressure (BP)-lowering and VaD-improving effects with the clinically used drug, lisinopril in DHRs while there was a positive correlation between the doses. Therefore, this bioactive compound may be useful for developing functional foods, thereby extending the application of equol-containing crops.

Topics: Acetylcholinesterase; Animals; Blood Pressure; Body Weight; Brain; Catalase; Dementia, Vascular; Desoxycorticosterone Acetate; Disease Models, Animal; Dose-Response Relationship, Drug; Equol; Glutathione Peroxidase; Hypertension; Malondialdehyde; Memory, Short-Term; Nitric Oxide; Oxidative Stress; Physical Conditioning, Animal; Phytoestrogens; Rats; Renin-Angiotensin System; Superoxide Dismutase; Swimming

Curcuma comosa Roxb. (C. comosa) or Wan Chak Motluk, Zingiberaceae family, has been used in Thai traditional medicine for the treatment of gynecological problems and inflammation.. This study aimed to investigate the therapeutic potential of C. comosa by determining the changes in the lipid profiles in the ovariectomized rats, as a model of estrogen-deficiency-induced hyperlipidemia, after treatment with different components of C. comosa using an untargeted lipidomics approach.. Lipids were extracted from the serum of adult female rats subjected to a sham operation (SHAM; control), ovariectomy (OVX), or OVX with 12-week daily doses of estrogen (17β-estradiol; E. Levels of five classes of lipids (ceramide, ceramide-1-phosphate, sphingomyelin, 1-O-alkenyl-lysophosphatidylethanolamine and lysophosphatidylethanolamine) were elevated in the OVX rats compared to those in the SHAM rats, while the monoacylglycerols and triacylglycerols were decreased. The E. The findings suggest the potential beneficial effects of C. comosa on preventing the increased ceramide levels in OVX rats, a possible cause of metabolic disturbance under estrogen deficiency. Overall, the results demonstrated the power of untargeted lipidomics in discovering disease-relevant biomarkers, as well as evaluating the effectiveness of treatment by C. comosa components (DPHD, extract or powder) as utilized in Thai traditional medicine, and also providing scientific support for its folklore use.

Topics: Animals; Biomarkers; Chromatography, Liquid; Curcuma; Diarylheptanoids; Discriminant Analysis; Disease Models, Animal; Estradiol; Estrogen Replacement Therapy; Ethanol; Female; Heptanol; Hyperlipidemias; Hypolipidemic Agents; Lipids; Metabolomics; Multivariate Analysis; Ovariectomy; Phytoestrogens; Phytotherapy; Plant Extracts; Plants, Medicinal; Powders; Rats, Sprague-Dawley; Rhizome; Solvents; Tandem Mass Spectrometry

This study aimed to ascertain the optimal range of red clover dry extracts (RC) and dried pomegranate concentrate powder (PCP) to induce anti-climacteric effects. Thus, the dose ranges showing protective effect of mixed formulae consisting of RC and PCP were examined in ovariectomized mice. At 28 days after bilateral ovariectomy (OVX), mixed herbal compositions (RC:PCP = 1:1, 1:2, 1:4, 1:8, 2:1, 4:1, and 8:1) were administered orally, at 120 mg/kg once daily for 84 days. We evaluated that RC and PCP mixture attenuate OVX-caused obesity, hyperlipidemia, hepatic steatosis, and osteoporosis. Compared to OVX-induced control mice, body weight and abdominal fat weight in OVX-induced mice were significantly decreased, concomitantly with increase of uterus weight by RC:PCP mixture. Additionally, significant increases in serum estradiol levels were observed in all RC:PCP-treated mice. RC:PCP mixture also showed protective effect against OVX-induced hyperlipidemia, hepatic steatosis. Total body and femur mean bone mineral density (BMD), osteocalcin, bALP contents were effectively increased by RC:PCP mixture. Taken together, RC:PCP mixture (2:1, 1:1, and 4:1) has remarkable protective effects against the changes induced by OVX. In particular, RC:PCP mixture (2:1) shows the strongest effect and may be considered as a potential protective agent against climacteric symptoms.

Topics: Animals; Animals, Outbred Strains; Anti-Obesity Agents; Biomarkers; Bone Density Conservation Agents; Dietary Supplements; Disease Models, Animal; Fatty Liver; Female; Fruit; Humans; Hyperlipidemias; Lipid Regulating Agents; Lythraceae; Mice; Obesity; Osteoporosis, Postmenopausal; Phytoestrogens; Plant Extracts; Plant Leaves; Specific Pathogen-Free Organisms; Trifolium

Obesity emerged as the major risk factor for metabolic syndrome. Postmenopausal women are more prone to develop obesity than premenopausal women. The absence of safe and effective conventional treatments for postmenopausal obesity has changed the focus to natural products as alternative remedy. We investigated the molecular basis of the effect of soy isoflavones (SIFs) on hypertriglyceridemia and hepatic steatosis in an animal model of postmenopausal obesity. Ovariectomized (OVX) and sham-operated Wistar rats were fed with high-fat diet (HFD) and normal diet for 8 weeks with and without SIF extract (150mg/kg body weight/day). Both OVX and HFD per se and when combined caused hypertriglyceridemia, hypercholesterolemia and atherogenic lipid profile. Proteomic studies revealed that both OVX and HFD caused overexpression of hepatic lipogenic proteins, such as LXR, SREBP1, PPARγ, ACC and FAS, in association with reduced expression of lipolytic proteins, such as FXR, PPARα, insig2 and SHP. Histological analysis showed fat accumulation and morphological abnormalities in the liver of OVX and HFD rats. All these metabolic derangements were further augmented when OVX was followed by HFD. In conclusion, these findings suggest that there was a synergism in the development of deranged lipid metabolism with the coexistence of postmenopausal state and the intake of fat-rich diet. SIF extract markedly alleviated the derangement of lipid metabolism suggesting the use of this natural phytoestrogen as a strategy for relieving dyslipidemia and hepatic steatosis associated with the postmenopausal women.

Topics: Animals; Biomarkers; Diet, High-Fat; Dietary Supplements; Disease Models, Animal; Dyslipidemias; Female; Humans; Hypertriglyceridemia; Isoflavones; Lipid Metabolism; Liver; Metabolic Syndrome; Non-alcoholic Fatty Liver Disease; Obesity; Organ Size; Osteoporosis, Postmenopausal; Ovariectomy; Phytoestrogens; Random Allocation; Rats, Wistar; Soy Foods

The development of atherosclerosis is closely related to excessive endoplasmic reticulum stress (ERs). Equol reportedly protects against cardiovascular disease; however, the underlying mechanism for this protection remains unknown. Herein, the mechanisms contributing to the atheroprotective effect of equol were addressed using apolipoprotein E knockout (apoE-/-) mice fed a high-fat diet (HFD) with or without equol. Equol intervention reduced atherosclerotic lesions in the aorta in HFD-fed apoE-/- mice. Plasma lipid analysis showed that equol intervention reduced triglycerides, total cholesterol and LDL-cholesterol and increased HDL-cholesterol. Additionally, equol administration decreased lipid accumulation in the liver. Simultaneously, equol treatment inhibited cell apoptosis induced by t-BHP and thapsigargin in human umbilical vein endothelial cells (HUVECs). Furthermore, equol treatment attenuated palmitate, t-BHP or thapsigargin-induced upregulation of ER stress markers, including p-PERK, p-eIF2α, GRP78, ATF6 and CHOP proteins expression. The same tendency was also observed in aortic lysates in apoE-/- mice fed with equol plus HFD compared with HFD alone. Moreover, equol treatment dose dependently activated the Nrf2 signaling pathway under oxidative stress. Additionally, elevation of Nrf2 induction was found in aortic lysates in apoE-/- mice fed with a HFD diet containing equol compared with a HFD diet without equol. Importantly, Nrf2 siRNA interference induced CHOP and attenuated the effect of equol to inhibit t-BHP mediated CHOP induction, furthermore, abrogated cell apoptosis induced by t-BHP, suggesting a role for Nrf2 in the protective effect of equol in HUVECs. Collectively, these findings implicate that the improvement of atherosclerosis by equol through attenuation of ER stress is mediated, at least in part, by activating the Nrf2 signaling pathway.

Topics: Activating Transcription Factor 6; Animals; Aorta; Apolipoproteins E; Apoptosis; Atherosclerosis; Cholesterol, HDL; Cholesterol, LDL; Diet, High-Fat; Disease Models, Animal; eIF-2 Kinase; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Equol; Eukaryotic Initiation Factor-2; Gene Expression Regulation; Heat-Shock Proteins; Human Umbilical Vein Endothelial Cells; Humans; Liver; Mice; Mice, Knockout; NF-E2-Related Factor 2; Palmitic Acid; Phytoestrogens; Signal Transduction; Thapsigargin; Transcription Factor CHOP; Triglycerides

Post-menopausal osteoporosis has long been treated and prevented by estrogen replacement therapy (ERT). Despite its effectiveness, ERT is associated with serious adverse effects. Labisia pumila var. alata (LP) is a herb with potential as an alternative agent to ERT due to its phytoestrogenic, antioxidative and anti-inflammatory effects on bone. This study aimed to determine the effects of LP supplementation on bone biomechanical strength of postmenopausal osteoporosis rat model.. Ninety-six female Sprague-Dawley rats aged 4 to 5 months old were randomly divided into six groups; six rats in the baseline group (BL) and eighteen rats in each group of; Sham- operated (Sham), ovariectomised control (OVXC) and ovariectomised with daily oral gavages of Premarin at 64.5 μg/kg (ERT), LP at 20 mg/kg (LP20) and LP at 100 mg/kg (LP100) respectively. These groups were subdivided into three, six and nine weeks of treatment periods. Rats in BL group were euthanized before the start of the study, while other rats were euthanized after completion of their treatments. Femora were dissected out for biomechanical strength analysis using Instron Universal Model 5848 Micro Tester.. OVXC group showed deterioration in the bone biomechanical strength with time. Both ERT and LP supplemented rats showed improvements in bone strength parameters such as maximum load, displacement, stiffness, stress, and Young Modulus. The most improved bone strength was seen in rats given LP at the dose of 100 mg/kg for nine weeks.. LP supplementation at 100 mg/kg was more effective than ERT in reversing ovariectomy-induced bone biomechanical changes.

Topics: Animals; Biomechanical Phenomena; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Femur; Humans; Osteoporosis, Postmenopausal; Ovariectomy; Phytoestrogens; Phytotherapy; Plant Extracts; Primulaceae; Rats, Sprague-Dawley; Stress, Mechanical

Breast cancer prevention efforts are focused increasingly on potentially beneficial dietary modifications due to their ease of implementation and wide acceptance. Secoisolariciresinol diglucoside (SDG) is a lignan found in high concentration in flaxseed that may have selective estrogen receptor modulator-like effects resulting in antiestrogenic activity in a high estrogen environment. In parallel with a human phase II prevention trial, female ACI rats (n = 8-10/group) received 0, 10, or 100 ppm SDG in the feed. The 100 ppm SDG treatment produced similar blood lignan levels as those observed in our human pilot study. Mammary and ovarian cancer progression were induced using local ovarian DMBA treatment and subcutaneous sustained release 17β-estradiol administered starting at 7 weeks of age. Mammary gland and ovarian tissues were collected at 3 mo after initiation of treatment and examined for changes in epithelial cell proliferation (Ki-67, cell counts), histopathology, and dysplasia scores, as well as expression of selected genes involved in proliferation, estrogen signaling, and cell adhesion. Treatment with SDG normalized several biomarkers in mammary gland tissue (dysplasia, cell number, and expression of several genes) that had been altered by carcinogen. There is no indication that SDG promotes preneoplastic progression in the ovarian epithelium.

Topics: Animals; Biomarkers, Tumor; Breast Neoplasms; Butylene Glycols; Cell Adhesion; Disease Models, Animal; Disease Progression; Drug Evaluation, Preclinical; Estrogen Receptor Modulators; Female; Flax; Glucosides; Ki-67 Antigen; Ovarian Neoplasms; Phytoestrogens; Precancerous Conditions; Rats; Rats, Inbred ACI; Seeds

The aim of this study was to assess the effects of genistein (G) and daidzein (D) on the histological, hormonal, and functional parameters of the pituitary-ovarian axis in middle-aged female rats, and to compare these effects with the effects of estradiol (E), commonly used in the prevention and treatment of menopausal symptoms. Middle-aged (12 month old) Wistar female rats subcutaneously received 35 mg/kg of G, or 35 mg/kg of D, or 0.625 mg/kg of E every day for 4 weeks. Each of the treated groups had a corresponding control group. An intact control group was also established. G and D did not change the intracellular protein content within gonadotropic and lactotropic cells, but vacuolization was observed in all the cell types. In contrast, E caused an inhibition of gonadotropic and stimulation of lactotropic cells. Also, ovaries of middle-aged female rats exposed to G or D have more healthy primordial and primary follicles and less atretic follicles. E treatment in the ovaries had a mostly negative effect, which is reflected by the increased number of atretic follicles in all tested classes. G and D provoked decrease in CuZnSOD and CAT activity, while E treatment increased MnSOD and decreased CuZnSOD and GSHPx activity. All the treatments increased serum estradiol and decreased testosterone levels, while D and E increased the serum progesterone level. In conclusion, soy phytoestrogens exhibited beneficial effects on pituitary-ovarian function in middle-aged female rats, as compared to estradiol.

Topics: Animals; Antioxidants; Disease Models, Animal; Drug Evaluation, Preclinical; Estrogen Replacement Therapy; Female; Genistein; Hormones; Isoflavones; Menopause; Ovary; Phytoestrogens; Pituitary Gland; Rats, Wistar

BRF2 is a transcription factor required for synthesis of a small group of non-coding RNAs by RNA polymerase III. Overexpression of BRF2 can transform human mammary epithelial cells. In both breast and lung cancers, the BRF2 gene is amplified and overexpressed and may serve as an oncogenic driver. Furthermore, elevated BRF2 can be independently prognostic of unfavorable survival. Dietary soy isoflavones increase metastasis to lungs in a model of breast cancer and a recent study reported significantly increased cell proliferation in breast cancer patients who used soy supplementation. The soy isoflavone daidzein is a major food-derived phytoestrogen that is structurally similar to estrogen. The putative estrogenic effect of soy raises concern that high consumption of soy foods by breast cancer patients may increase tumor growth.. Expression of BRF2 RNA and protein was assayed in ER-positive or -negative human breast cancer cells after exposure to daidzein. We also measured mRNA stability, promoter methylation and response to the demethylating agent 5-azacytidine. In addition, expression was compared between mice fed diets enriched or deprived of isoflavones.. We demonstrate that the soy isoflavone daidzein specifically stimulates expression of BRF2 in ER-positive breast cancer cells, as well as the related factor BRF1. Induction is accompanied by increased levels of non-coding RNAs that are regulated by BRF2 and BRF1. Daidzein treatment stabilizes BRF2 and BRF1 mRNAs and selectively decreases methylation of the BRF2 promoter. Functional significance of demethylation is supported by induction of BRF2 by the methyltransferase inhibitor 5-azacytidine. None of these effects are observed in an ER-negative breast cancer line, when tested in parallel with ER-positive breast cancer cells. In vivo relevance is suggested by the significantly elevated levels of BRF2 mRNA detected in female mice fed a high-isoflavone commercial diet. In striking contrast, BRF2 and BRF1 mRNA levels are suppressed in matched male mice fed the same isoflavone-enriched diet.. The BRF2 gene that is implicated in cancer can be induced in human breast cancer cells by the isoflavone daidzein, through promoter demethylation and/or mRNA stabilization. Dietary isoflavones may also induce BRF2 in female mice, whereas the converse occurs in males.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Disease Models, Animal; DNA Methylation; Female; Humans; Isoflavones; Male; Neoplasm Proteins; Phytoestrogens; Promoter Regions, Genetic; Proto-Oncogene Mas; RNA, Messenger; RNA, Neoplasm; TATA-Binding Protein Associated Factors; Transcription Factor TFIIIB

To investigate the effect of either genistein, or exercise, or both, on parameters that are indicators of cardiovascular health.. We investigated the effect of genistein treatment (300 mg genisten/kg body weight/day), or exercise training, or combined genistein and exercise training, for a period of 6 weeks on physical characteristics, cardiovascular plasma markers, blood pressure, aortic morphology, cardiac structure and oxidative stress in the ovariectomized (OVX) Sprague-Dawley rat. Comparisons were made with intact rats.. Ovariectomy (compared to intact) resulted in significant decreases in uterine weight (6-fold, p < 0.0001), insulin levels (4-fold, p = 0.0214), insulin/glucose ratio (3-fold, p = 0.0029), and tumor necrosis factor-α plasma levels (2-fold, p < 0.0001). Similarly, aortic blood pressure was significantly increased (by 8%, p < 0.0033) in OVX rats, without changes in aortic luminal or wall dimensions. Heart surface area was significantly increased (by 16%, p = 0.0160) in OVX rats and this was without changes in non-protein thiol levels (a marker of oxidative stress). Physical characteristics were not altered by treatment with genistein, or genistein with exercise, with the exception of increased uterine weight in OVX rats treated under these same conditions. There were no effects of genistein or exercise on indices of blood pressure and aortic morphology in the OVX rat. However, right ventricular nuclei count was reduced in sedentary genistein-treated rats compared to non-treated control OVX rats.. Our results indicate that administration of genistein at this dose, treadmill running, or the combination of both, are not associated with any improvement in cardiovascular function and structure, and risk factors in an ovariectomy model of postmenopause.

Topics: Animals; Blood Pressure; Cardiovascular Diseases; Dietary Supplements; Disease Models, Animal; Female; Genistein; Heart Rate; Ovariectomy; Physical Conditioning, Animal; Phytoestrogens; Rats; Rats, Sprague-Dawley; Risk Factors

Resveratrol (Res), a polyphenol that is abundant in many medicinal plants and is a selective oestrogen receptor modulator, exhibits multiple biological activities. In the present study, we determined whether Res prevents oestrogen deficiency-induced osteopenia and whether Res administration decreases pathological changes in the endometrium and lumen of the uterus compared with oestradiol replacement therapy (ERT). A total of sixty 3-4-month-old female Wistar rats were randomly divided into a sham-operated group (Sham) and five ovariectomy (OVX) subgroups, i.e. OVX rats as a control group (OVX); OVX rats receiving oestradiol valerate (ERT, 0·8 mg/kg); and OVX rats receiving Res 20, 40 and 80 mg/kg. Daily oral administration was initiated at week 2 after OVX for 12 weeks. A dose-response difference was observed in the effects of Res on bone mineral density (BMD) and trabecular microarchitecture. Only at the highest dose, bone loss was almost equivalent to that observed in the ERT group. The dose-response effects of Res on the biochemical parameters (alkaline phosphatase, IL-6, TNF-α and transforming growth factor-β1 concentrations in the serum as well as urinary Ca and P excretion) and the expressions of receptor activator of nuclear factor κB ligand (RANKL) and the RANKL:osteoprotegerin protein ratio in the femur were also observed. Furthermore, the thickening of the endometrium and the infiltration of lymphocytes were prevented in all the three Res-treated groups compared with the ERT group. In conclusion, Res treatment not only improves BMD and trabecular microarchitecture but also does not affect the uterus and Res might be a potential remedy for the treatment of postmenopausal osteoporosis.

Topics: Animals; Antioxidants; Biomarkers; Bone Density; Bone Density Conservation Agents; Dietary Supplements; Disease Models, Animal; Endometrial Hyperplasia; Endometrium; Estrogen Replacement Therapy; Female; Femur; Humans; Osteoporosis, Postmenopausal; Osteoprotegerin; Phytoestrogens; Random Allocation; RANK Ligand; Rats; Rats, Wistar; Resveratrol; Stilbenes; Time Factors

To evaluate the effects of soy isoflavone extract in the pro-oxidant/antioxidant balance in the uterus of ovariectomized rats.. Twenty 3-month-old adult female Wistar rats were divided into four equal groups: GI: sham-operated (estrous phase); GII: control ovariectomized rats; GIII: ovariectomized rats treated with genistein (50 μg/kg/day) by gavage; GIV: ovariectomized rats subcutaneously treated with estrogen (10 μg/kg/day). After 30 consecutive days of treatment, the rats were euthanized and the uterus removed. The distal thirds of the uterine horns were processed for histomorphometric analyses of endometrial and myometrial thicknesses and glandular area. Other regions of the uteri were kept in liquid nitrogen and subsequently processed for analysis of reactive species quantification (DCF), total antioxidant capacity (TAC) and lipid oxidation status (TBARS). Data were statistically analyzed by one-way ANOVA, complemented by the Tukey-Kramer test (p < 0.05).. GII and GIII exhibited lower endometrial thickness, glandular area and myometrial thickness than GI and GIV, while a higher myometrial thickness was observed in GIV compared with the other groups. Moreover, the isoflavone-treated group showed lower DCF and TBARS compared to GII, and also an improvement of TAC compared to GI and GIV. Despite the significant decrease in TBARS, no significant difference in DCF nor a decrease in TAC were observed in GIV when compared to GII.. Our data show that isoflavones improve antioxidant status and counteract oxidative stress, without promoting a trophic effect in the uterus of rats.

Topics: Animals; Disease Models, Animal; Drug Administration Routes; Estrogens; Female; Genistein; Glycine max; Isoflavones; Lipid Metabolism; Ovariectomy; Oxidative Stress; Phytoestrogens; Plant Extracts; Progesterone; Rats; Rats, Wistar; Treatment Outcome; Uterus

To examine effects of equol, the soy phytoestrogen metabolite, on gene expression in the monkey iliac artery.. A high fat/high cholesterol diet was fed to eight ovariectomized cynomolgus monkeys for 6.5 years. After biopsy of the left iliac artery, the animals were randomized to two treatment groups for 8 months; the treatment groups were equol (23.7 mg/100 g diet, n = 4) and vehicle (n = 4). The right iliac artery was removed at necropsy. Gene expression in the iliac arteries in response to equol was determined by DNA microarray. Comparison of atherosclerotic lesions and plasma lipids at pre-versus post-equol treatment time points and in vehicle versus equol treatment groups did not identify any significant differences. Despite the lack of effect of equol on these parameters, 59 genes were down-regulated and 279 were up-regulated in response to equol. Comparison of these data to previous work identified 10 genes regulated in opposite directions by equol compared to presence of atherosclerosis plaque (Menopause 2011; 18:1087-1095) and 55 genes differentially expressed in the same direction in response to both equol and estradiol (Eyster et al., Menopause 2014;21:143-152.).. Similar responses of genes to both equol and estradiol may reflect the extent to which equol serves as a natural selective estrogen receptor modulator in the arteries. Opposite responses of 10 genes to equol versus the presence of atherosclerosis implicates those genes in the potential protective effects of equol in arteries.

Topics: Animals; Atherosclerosis; Diet, High-Fat; Disease Models, Animal; Equol; Female; Gene Expression Regulation; Iliac Artery; Macaca fascicularis; Ovariectomy; Phytoestrogens; Plaque, Atherosclerotic; Time Factors

Millettia macrophylla Benth is a Cameroonian medicinal plant traditionally used to alleviate menopause-related problems. The methanol soluble fraction of this plant was shown to exhibit estrogenic effects in vitro in Human Embryonic kidney cells, and in vivo on ovariectomized rat following the classical uterotrophic assay. Since estrogens have been involved in bone remodeling process, the present study then aimed at evaluating bone loss preventive effects of the methanol soluble fraction of Millettia macrophylla (MM-met) in ovariectomized rat model.. Twenty-five healthy Wistar female rats (3-month-old) were randomly assigned to a sham-operated group and to four treated ovariectomized (OVX) groups. Treatments lasted 8 weeks and animals were sacrificed. The uterus, the femoral and the tibia bones of each animal were collected, weighed and fixed in 10% formalin for histological analysis.. Results showed that ovariectomy decreased uterine wet weight (p<0.01), induced body weight gain (p<0.01), decreased both femoral and tibia bone density and mineral content and increased alkaline phosphatase activity (p<0.05). E2V and MM-met treatments in general prevented bone mass loss and/or bone density loss. At all tested doses, MM-met induced a significant decrease of alkaline phosphatase activity (p<0.05). As observed with E2V, MM-met also induced a significant protective effect on bone, and this was indicated by an abundance of bone marrow in an almost intact trabecular network.. The overall results show that the methanol soluble fraction of Millettia macrophylla may prevent ovariectomy-induced bone mass loss and deterioration of the trabecular microarchitecture.

Topics: Alkaline Phosphatase; Animals; Body Weight; Bone Density; Bone Density Conservation Agents; Bone Marrow; Disease Models, Animal; Female; Femur; Leg Bones; Millettia; Organ Size; Osteoporosis; Ovariectomy; Phytoestrogens; Phytotherapy; Plant Extracts; Rats, Wistar; Tibia

Intermittent hypoxia was introduced to mimic obstructive sleep apnea-hypopnea syndrome (OSAHS) in rats. Then, bone mass, bone strength and bone turnover were evaluated, and the influence of genistein on bone mass reduction was investigated in these rats.. OSAHS animal model was established via chronic intermittent hypoxia, and genistein (2.5 mg/kg/day) was used to treat OSAHS rats. The bone mineral density (BMD), bone Histomorphometric indicators, bone biomechanics and expressions of genes related to bone formation and resorption (Runx2, Col I, ALP, Osteocalcin, OPG, RANKL and TRAP-5b) were measured after treatment.. The BMD in OSAHS+OVX group was significantly lower than that in OVX group (P<0.05). The BMD in OSAHS+OVX+Genistein group was markedly increased when compared with OSAHS+OVX group (P<0.05), accompanied by partial improvement of the OSAHS induced damage to the lumbar biomechanics. In OSAHS+OVX group, the expressions of Runx2, Col I, ALP and Osteocalcin were significantly reduced when compared with OVX group, and rats in OSAHS+OVX+Genistein group had significantly higher expressions of Runx2, Col I, ALP and Osteocalcin and reduced TRAP-5b expression as compared to OSAHS+OVX group (P<0.05).. Genistein can improve the reduction in bone mass and bone strength due to OSAHS in OVX rats, which may be attributed to the increase in bone formation and inhibition of bone resorption. Our findings suggest that genistein may be used to treat and prevent osteoporosis in postmenopausal women with OSAHS.

Topics: Animals; Biomechanical Phenomena; Bone Density; Bone Resorption; Collagen Type I; Core Binding Factor Alpha 1 Subunit; Disease Models, Animal; Female; Genistein; Hypoxia; Osteocalcin; Osteogenesis; Phytoestrogens; Rats; Rats, Sprague-Dawley; Sleep Apnea, Obstructive

Formononetin has shown a variety of pharmacologic properties including anti-inflammatory effect. In the present study, we analyzed the role of formononetin in acute lung injury induced by lipopolysaccharide (LPS) in mice. The cell counting in the bronchoalveolar lavage fluid (BALF) was measured. The animal lung edema degree was evaluated by wet/dry weight ratio. The superoxidase dismutase (SOD) activity and myeloperoxidase (MPO) activity was assayed by SOD and MPO kits, respectively. The levels of inflammatory mediators, tumor necrosis factor-α (TNF-α) and IL-6,were assayed by enzyme-linked immunosorbent assay method. Pathological changes of hung tissues were observed by HE staining. Peroxisome proliferator-activated receptor (PPAR)-γ gene expression was measured by real-time PCR. The data showed that treatment with the formononetin group markedly attenuated inflammatory cell numbers in the BALF, increased PPAR-γ gene expression and improved SOD activity and inhibited MPO activity. The histological changes of the lungs were also significantly improved by formononetin compared to LPS group. The results indicated that formononetin has a protective effect on LPS-induced acute lung injury in mice.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Edema; Inflammation; Interleukin-6; Isoflavones; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred BALB C; Peroxidase; Phytoestrogens; PPAR gamma; Pulmonary Edema; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Microglial activation has been implicated in various neurological disorders, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and HIV encephalopathy. Phytoestrogens have been shown to be neuroprotective in neurotoxicity models; however, their effect on microglia has not been well established. In the current study, we report that the soy phytoestrogens, genistein, daidzein, and coumestrol, decreased nitric oxide (NO) production induced by lipopolysaccharide (LPS) in the rat microglial cell line (HAPI). The levels of inducible NO synthase (iNOS) mRNA and protein expression were also reduced. Transcription factors known to govern iNOS expression including interferon regulatory factor-1 (IRF-1) and phosphorylated STAT1 were down regulated. These observations explain, at least in part, the inhibitory effect of phytoestrogens on NO production. The levels of monocyte chemoattractant protein-1 and interleukin-6 mRNA, proinflammatory chemokine and cytokine associated with various neurological disorders, were also reduced following LPS stimulation when HAPI cells were pretreated with phytoestrogens. Hence, genistein, daidzein, and coumestrol could serve as anti-inflammatory agents and may have beneficial effects in the treatment of neurodegenerative diseases.

Topics: Animals; Anti-Inflammatory Agents; Cell Line, Transformed; Chemokine CCL2; Disease Models, Animal; Gene Expression Regulation; Glycine max; Interferon Regulatory Factor-1; Interleukin-6; Lipopolysaccharides; Microglia; Neurodegenerative Diseases; Neuroprotective Agents; Nitric Oxide; Nitric Oxide Synthase Type II; Phytoestrogens; Rats; STAT1 Transcription Factor

Our recent developments have yielded a novel phytoestrogenic formulation, referred to as the phyto-β-SERM formulation, which exhibits an 83-fold binding selectivity for the estrogen receptor subtype β (ERβ) over ERα. Earlier studies indicate that the phyto-β-SERM formulation is neuroprotective and promotes estrogenic mechanisms in the brain while devoid of feminizing activity in the periphery. Further investigation in a mouse model of human menopause indicates that chronic exposure to the phyto-β-SERM formulation at a clinically relevant dosage prevents/alleviates menopause-related climacteric symptoms. This study assessed the efficacy, in an early intervention paradigm, of the phyto-β-SERM formulation in the regulation of early stages of physical and neurological changes associated with Alzheimer's disease (AD) in a female triple transgenic mouse model of AD. Results demonstrated that, when initiated prior to the appearance of AD pathology, a 9-month dietary supplementation with the phyto-β-SERM formulation promoted physical health, prolonged survival, improved spatial recognition memory, and attenuated amyloid-β deposition and plaque formation in the brains of treated AD mice. In comparison, dietary supplementation of a commercial soy extract preparation showed no effect on cognitive measures, although it appeared to have a positive impact on amyloid pathology. In overall agreement with the behavioral and histological outcomes, results from a gene expression profiling analysis offered insights on the underlying molecular mechanisms associated with the two dietary treatments. In particular, the data suggests that there may be a crosstalk between ERβ and glycogen synthase kinase 3 signaling pathways that could play a role in conferring ERβ-mediated neuroprotection against AD. Taken together, these results support the therapeutic potential of the phyto-β-SERM formulation for prevention and/or early intervention of AD, and warrants further investigations in human studies.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Animals; Disease Models, Animal; Estrogen Receptor beta; Female; Humans; Maze Learning; Memory Disorders; Mice; Mice, Transgenic; Mutation; Ovariectomy; Peptide Fragments; Phytoestrogens; Plaque, Amyloid; Presenilin-1; Recognition, Psychology; tau Proteins

Irritable bowel syndrome (IBS) often associated with psychological distress, is characterized by increased gut permeability and visceral sensitivity. In animals, stress increases intestinal paracellular permeability (IPP), visceral sensitivity and colonic proteolytic activity. Estradiol reduces IPP and affects visceral sensitivity in non-stressed ovariectomized rats, but whether estrogens affect stress-induced hyperpermeability and hypersensitivity in cyclic females remains unclear. We aimed to evaluate (i) the effects of a phytoestrogen-rich soy germ fermented ingredient (SG) on visceral hypersensitivity, hyperpermeability and other symptoms in stressed intact female rats, (ii) the mechanisms of action involved on the basis of both estrogenic and protease inhibitor activities of SG.. Female rats received orally for 15-d either SG, 17β-estradiol benzoate (EB), or vehicles, with or without the estrogen receptor (ER) antagonist ICI182.780 before stress. Visceral sensitivity, IPP, faecal proteolytic activity, plasma corticosterone, rat mast cell protease II immunostaining, and occludin expression were assessed.. Stress increased IPP (concomitantly to a drop in occludin expression), visceral sensitivity, faecal proteolytic activity and plasma corticosterone. Similarly to EB, SG prevented the stress-induced hyperpermeability, and hypersensitivity, without changes in plasma corticosterone. SG inhibited the increase in faecal proteolytic activity, enhanced occludin expression, and reduced the colonic mast cell density. All SG effects, except decrease on faecal proteolytic activity, were blocked by ICI182.780.. A 2-wk oral treatment with SG prevented the stress-induced hyperpermeability and visceral hypersensitivity in cyclic rats through ER activation, and blocked the increase in colonic proteolytic activity, suggesting that SG can be promising in IBS management.

Topics: Animals; Disease Models, Animal; Estradiol; Estrogen Receptor Modulators; Feces; Female; Fermentation; Fulvestrant; Gastrointestinal Tract; Germination; Glycine max; Irritable Bowel Syndrome; Permeability; Phytoestrogens; Protease Inhibitors; Proteolysis; Rats; Rats, Wistar; Receptors, Estrogen; Seeds; Soy Foods; Stress, Physiological; Stress, Psychological

This study was designed to develop a bioactive scaffold to enhance bone defect repair in steroid-associated osteonecrosis (SAON). Icaritin, a metabolite of the herb Epimedium, has been identified as an angiogenic and osteogenic phytomolecule. Icaritin was homogenized into poly lactic-co-glycolic acid/tricalcium phosphate (PLGA/TCP) to form an icaritin-releasing porous composite scaffold (PLGA/TCP/icaritin) by fine-spinning technology. In vitro, high performance liquid chromatography was used to determine the release of icaritin during degradation of PLGA/TCP/icaritin. The osteogenic effects of PLGA/TCP/icaritin were evaluated using rat bone marrow mesenchymal stem cells (BMSCs). In vivo, the osteogenic effect of PLGA/TCP/icaritin was determined within a bone tunnel after core decompression in SAON rabbits and angiography within scaffolds was examined in rabbit muscle pouch model. In vitro study confirmed the sustainable release of icaritin from PLGA/TCP/icaritin with the bioactive scaffold promoting the proliferation and osteoblastic differentiation of rat BMSCs. In vivo study showed that PLGA/TCP/icaritin significantly promoted new bone formation within the bone defect after core decompression in SAON rabbits and enhanced neovascularization in the rabbit muscle pouch experiment. In conclusion, PLGA/TCP/icaritin is an innovative local delivery system that demonstrates sustainable release of osteogenic phytomolecule icaritin enhancing bone repair in an SAON rabbit model. The supplement of scaffold materials with bioactive phytomolecule(s) might improve treatment efficiency in challenging orthopedic conditions.

Topics: Animals; Bone Marrow Cells; Calcium Phosphates; Cells, Cultured; Disease Models, Animal; Femoral Fractures; Flavonoids; Fracture Healing; Lactic Acid; Male; Neovascularization, Physiologic; Osteogenesis; Osteonecrosis; Phytoestrogens; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Rabbits; Rats; Tissue Engineering; Tissue Scaffolds

Several studies have shown that soy isoflavones have estrogen-like activities and might constitute an alternative to hormone replacement treatment. The present study investigated the effects of soy isoflavones alone and combined with vitamin D3 on prevention of bone loss.. Sprague-Dawley rats were sham-operated (n = 8) or ovariectomized (OVX; n = 40), and then the OVX rats were randomly assigned to five groups that were untreated or treated for 14 wk with vitamin D3, 17β-estradiol, soy isoflavone extract (SIE), or vitamin D3 plus SIE. The effects of the isoflavones and 1α,25(OH)(2)D(3) on cultured osteoblasts and osteoclasts also were investigated.. In OVX rats, the bone mineral density and trabecular bone volume loss were improved by 17β-estradiol, SIE, or SIE plus vitamin D3 treatment. SIE treatment was more effective than vitamin D3 or 17β-estradiol in inhibiting increases in serum tumor necrosis factor-α levels and osteoblast osteoprotegerin expression. SIE plus vitamin D3 was more effective in increasing osterix expression than each alone. Bone cell cultures showed that the isoflavones induced preosteoblasts to differentiate into osteoblasts and increased osteoblast mineralization. Isoflavones inhibited preosteoclasts and osteoclast proliferation and decreased osteoclast resorption. The combination of isoflavones plus 1α,25(OH)(2)D(3) showed additive effects on the increase in cell proliferation of cultured preosteoblasts.. Treatment with soy isoflavones might be an alternative to hormone replacement therapy in decreasing bone loss from postmenopausal estrogen deficiency. In addition, there are further effects on increasing transcription factor osterix expression and preosteoblast proliferation when these were combined with vitamin D3.

Topics: Alkaline Phosphatase; Animals; Bone Density; Cholecalciferol; Disease Models, Animal; Drug Synergism; Estradiol; Female; Glycine max; Humans; Interleukin-1beta; Isoflavones; Osteoblasts; Osteocalcin; Osteoclasts; Osteogenesis; Osteoporosis; Ovariectomy; Phytoestrogens; Rats; Rats, Sprague-Dawley; RNA, Messenger; Tumor Necrosis Factor-alpha

Chronic intermittent hypoxia (CIH) is a frequent feature of OSAHS. The present study was designed to evaluate the effects of genistein and estrogen on genioglossus contractile and regeneration properties in CIH rats and investigate the involvement of HIF-1α.. Ovariectomized female rats were exposed to CIH for 5 weeks. Genistein and estrogen were administered by intraperitoneal injection. The genioglossus myoblasts of rat were also isolated and cultured in vitro, and the HIF-1α shRNA lentivirus was used.. Muscle fatigue resistance and myogenic regeneration were significantly decreased after CIH but were partially reversed by estrogen and genistein treatment. The effect of estrogen was more powerful than that of genistein. Compared with control group, RT-PCR and western blotting showed higher levels of HIF-1α mRNA and protein in the CIH group, but estrogen and genistein treatment reduced the levels of HIF-1α mRNA and protein in rats exposed to CIH. In genioglossus myoblasts, the expression of HIF-1α was up-regulated under hypoxia rather than normoxia and decreased over time under both hypoxia and normoxia during myogenic differentiation. HIF-1α knockdown relieved myogenesis inhibition under hypoxia.. We concluded that genistein and estrogen may inhibit the overexpression of HIF-1α induced by CIH and improve the endurance and regeneration of the genioglossus muscle.

Topics: Animals; Cells, Cultured; Disease Models, Animal; Estrogens; Female; Gene Knockdown Techniques; Gene Silencing; Genistein; Hypoxia-Inducible Factor 1, alpha Subunit; Injections, Intraperitoneal; Lentivirus; Muscle Contraction; Muscle Development; Muscle Fatigue; Myoblasts, Skeletal; MyoD Protein; Myogenic Regulatory Factor 5; Myosin Heavy Chains; Ovariectomy; Pharyngeal Muscles; Phytoestrogens; Random Allocation; Rats; Rats, Sprague-Dawley; Regeneration; RNA, Small Interfering; Sleep Apnea, Obstructive

Estrogen deficiency after menopause results in rapid bone loss, predisposing women to osteoporotic fractures. Genistein, a phytoestrogen present in high concentrations in soy, is an ingredient in dietary supplements aggressively marketed for bone health. However, in a recent long-duration clinical trial in postmenopausal women, the efficacy of soy extracts in reducing bone loss was disappointing. To better understand the failure of soy extracts to consistently induce a robust skeletal response in women, we investigated the long-term (5 mo) efficacy of genistein, administered as a daily oral supplement, (1) in preventing cancellous bone loss in skeletally mature virgin Long-Evans rats ovariectomized at 7 months of age and (2) in improving cancellous bone mass and architecture in aged retired-breeder rats ovariectomized at 16 or 22 months of age.. Rats within each age group were randomly assigned into one of three treatment groups (n = 7-12 rats/group): (1) vehicle control, (2) genistein 485 μg/day, or (3) genistein 970 μg/day, resulting in mean (SE) serum genistein levels of 0.18 (0.10), 0.76 (0.15), and 1.48 (0.31) μM, respectively. Total tibia bone mass and density were evaluated using dual-energy x-ray absorptiometry, whereas cancellous bone mass and architecture in the tibial metaphysis, as well as cortical bone mass and architecture in the tibial diaphysis, were evaluated by micro-CT.. Oral genistein administered as a dietary supplement did not influence the cumulative effects of ovariectomy, aging, and/or reproductive history on cancellous and cortical bone mass and architecture.. Serum levels of genistein similar to those in women consuming a high-soy diet are ineffective in preventing or treating bone loss in rat models for postmenopausal osteoporosis.

Topics: Aging; Animals; Bone Density; Dietary Supplements; Disease Models, Animal; Female; Genistein; Humans; Osteoporosis, Postmenopausal; Ovariectomy; Phytoestrogens; Rats; Rats, Long-Evans; Reproduction; Tibia

Phytoestrogens have been implicated as promising therapeutic agents to treat the vascular impairment seen in menopausal women. The present study investigated the long-term effects of phytoestrogens from Curcuma comosa Roxb. on vascular relaxation of isolated thoracic aorta from ovariectomized (OVX) rats. Treatment of OVX rats for 12 weeks with C. comosa powder, hexane extract, and a novel phytoestrogen, diarylheptanoid-D3, [(3R)-1,7-diphenyl-(4E,6E)-4,6-heptadien-3-ol] prevented impairment of the endothelium-dependent relaxation response to acetylcholine in OVX, but not the endothelium-denude aortic ring relaxation in response to sodium nitroprusside. These data suggest that the vascular relaxation effect of C. comosa is mediated via endothelial cells. Treatment with D3 also increased endothelial nitric oxide synthase (eNOS) and estrogen receptor-α (ERα) protein expression in the aorta of OVX rats and suppressed elevated tumor necrosis factor-α (TNF-α) expression in OVX aortic rings. These results indicate that C. comosa treatment prevents impairment of vascular relaxation in estrogen-deficient animals via the ER-eNOS pathway as well as through its ability to promote an anti-inflammatory response.

Topics: Animals; Aorta, Thoracic; Curcuma; Disease Models, Animal; Female; Humans; Menopause; Ovariectomy; Phytoestrogens; Plant Extracts; Rats; Rats, Sprague-Dawley; Time Factors; Vasodilation

Genistein, a major phytoestrogen of soy, is considered a potential drug for the prevention and treatment of post-menopausal osteoporosis. Mounting evidence suggested a positive correlation between genistein consumption and bone health both in vivo and in vitro. Earlier studies have revealed that genistein acted as a natural estrogen analogue which activated estrogen receptor and exerted anti-osteoporotic effect. However, it remains unclear whether PTH, the most crucial hormone that regulates mineral homeostasis, participates in the process of genistein-mediated bone protection. In the present study, we compared the therapeutic effects between genistein and nilestriol and investigated whether PTH and its specific receptor PTHR1 altered in response to genistein-containing diet in the animal model of ovariectomy. Our results showed that genistein administration significantly improved femoral mechanical properties and alleviates femoral turnover. Genistein at all doses (4.5 mg/kg, 9.0 mg/kg and 18.0 mg/kg per day, respectively) exerted improved bending strength and b-ALP limiting effects than nilestriol in the present study. However, genistein administration did not exert superior effects on bone protection than nilestriol. We also observed circulating PTH restoration in ovariectomized rats receiving genistein at the dose of 18 mg/kg per day. Meanwhile, PTHR1 abnormalities were attenuated in the presence of genistein as confirmed by RT-PCR, Western blot and immunohistochemistry. These findings strongly support the idea that besides serving as an estrogen, genistein could interact with PTH/PTHR1, causing a superior mineral restoring effect than nilestriol on certain circumstance. In conclusion, our study reported for the first time that the anti-osteoporotic effect of genistein is partly PTH/PTHR1-dependent. Genistein might be a potential option in the prevention and treatment of post-menopausal osteoporosis with good tolerance, more clinical benefits and few undesirable side effects.

Topics: Alkaline Phosphatase; Animals; Bone Density; Creatinine; Disease Models, Animal; Estriol; Female; Femur; Genistein; Humans; Kidney; Osteoporosis, Postmenopausal; Ovariectomy; Parathyroid Hormone; Phytoestrogens; Protective Agents; Quinestrol; Rats; Rats, Sprague-Dawley; Receptor, Parathyroid Hormone, Type 1; Tensile Strength

While peripheral insulin resistance is common during obesity and aging in mice and people, the progression to type 2 diabetes (T2D) is largely due to loss of β-cell mass and function through apoptosis. We recently reported that genistein, a soy derived isoflavone, can improve glycemic control and β-cell function in insulin-deficient diabetic mice. However, whether it can prevent β-cell loss and diabetes in T2D mice is unknown. Our current study aimed to investigate the effect of dietary supplemented genistein in a nongenetic T2D mouse model. Nongenetic, middle-aged obese diabetic mice were generated by high fat diet and a low dose of streptozotocin injection. The effect of dietary supplementation of genistein on glycemic control and β-cell mass and function was determined. Dietary intake of genistein (250 mg·kg(-1) diet) improved hyperglycemia, glucose tolerance, and blood insulin level in obese diabetic mice, whereas it did not affect body weight gain, food intake, fat deposit, plasma lipid profile, and peripheral insulin sensitivity. Genistein increased the number of insulin-positive β-cell in islets, promoted islet β-cell survival, and preserved islet mass. In conclusion, dietary intake of genistein could prevent T2D via a direct protective action on β-cells without alteration of periphery insulin sensitivity.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Dietary Supplements; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Genistein; Glucose Tolerance Test; Hyperglycemia; Insulin; Insulin-Secreting Cells; Lipids; Male; Mice; Mice, Inbred C57BL; Phytoestrogens

We compared the effects of the S-enantiomer and racemic forms of equol on bone using ovariectomized (OVX) mice. Femoral bone mineral density and bone strength decreased in the OVX mice, but not in OVX mice administered 0.5 mg/d S-equol. This, however, did not hold for racemic equol. Serum and urine S-equol concentrations were higher in the mice administered S-equol than in those administered racemic equol. These results suggest that the inhibitory effects of S-equol on bone fragility in OVX mice are greater than those of racemic equol.

Topics: Animals; Bone Density; Chromatography, High Pressure Liquid; Disease Models, Animal; Equol; Female; Femur; Humans; Mice; Osteoporosis; Osteoporotic Fractures; Ovariectomy; Phytoestrogens; Stereoisomerism

Pretreatment with a phytoestrogen genistein has been shown to attenuate the development of pulmonary hypertension (PH). Because PH is not always diagnosed early, we examined whether genistein could also reverse preexisting established PH and prevent associated right heart failure (RHF). PH was induced in male rats by 60 mg/kg of monocrotaline. After 21 days, when PH was well established, rats received daily injection of genistein (1 mg/kg per day) for 10 days or were left untreated to develop RHF by day 30. Effects of genistein on human pulmonary artery smooth muscle cell and endothelial cell proliferation and neonatal rat ventricular myocyte hypertrophy were assessed in vitro. Severe PH was evident 21 days after monocrotaline, as peak systolic right ventricular pressure increased to 66.35±1.03 mm Hg and right ventricular ejection fraction reduced to 41.99±1.27%. PH progressed to RHF by day 30 (right ventricular pressure, 72.41±1.87 mm Hg; RV ejection fraction, 29.25±0.88%), and mortality was ≈75% in RHF rats. Genistein therapy resulted in significant improvement in lung and heart function as right ventricular pressure was significantly reduced to 43.34±4.08 mm Hg and right ventricular ejection fraction was fully restored to 65.67±1.08% similar to control. Genistein reversed PH-induced pulmonary vascular remodeling in vivo and inhibited human pulmonary artery smooth muscle cell proliferation by ≈50% in vitro likely through estrogen receptor-β. Genistein also reversed right ventricular hypertrophy (right ventricular hypertrophy index, 0.35±0.029 versus 0.70±0.080 in RHF), inhibited neonatal rat ventricular myocyte hypertrophy, and restored PH-induced loss of capillaries in the right ventricle. These improvements in cardiopulmonary function and structure resulted in 100% survival by day 30. Genistein restored PH-induced downregulation of estrogen receptor-β expression in the right ventricle and lung. In conclusion, genistein therapy not only rescues preexisting severe PH but also prevents the progression of severe PH to RHF.

Topics: Animals; Capillaries; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Estrogen Receptor beta; Genistein; Glycine max; Heart Failure; Humans; Hypertension, Pulmonary; In Vitro Techniques; Male; Phytoestrogens; Pulmonary Artery; Rats; Rats, Sprague-Dawley; Treatment Outcome

The objective of this study was to evaluate the effects of genistein and moderate intensity exercise on Achilles tendon collagen and cross-linking in intact and ovariectomized (OVX) female Sprague-Dawley rats. Rats were separated into eight groups (n = 9/group): intact or OVX, treadmill exercised or sedentary, genistein-treated (300 mg/kg/day) or vehicle. After 6 weeks, tendons were assayed for the collagen-specific amino acid hydroxyproline and hydroxylyslpyridinoline (HP). Collagen content was not influenced by exercise (P = 0.40) but was lower (P < 0.001) in OVX-vehicle rats compared with intact vehicle rats (OVX: 894 ± 35 μg collagen/mg dry weight; intact: 1185 ± 72 μg collagen/mg dry weight). In contrast, collagen content in OVX rats treated with genistein was greater (P = 0.010, 1198 ± 121 μg collagen/mg dry weight) when compared with untreated rats and was not different from intact rats (P = 0.89). HP content was lower in OVX genistein-treated rats when compared with intact genistein-treated rats, but only within the sedentary animals (P = 0.05, intact-treated: 232 ± 39 mmol/mol collagen; OVX-treated: 144 ± 21 mmol/mol collagen). Our findings suggest that ovariectomy leads to a reduction in tendon collagen, which is prevented by genistein. HP content, however, may not have increased in proportion to the addition of collagen. Genistein may be useful for improving tendon collagen content in conditions of estrogen deficiency.

Topics: Achilles Tendon; Animals; Collagen; Disease Models, Animal; Female; Genistein; Ovariectomy; Physical Conditioning, Animal; Phytoestrogens; Rats; Rats, Sprague-Dawley; Statistics as Topic

Postmenopausal women are at higher risk for obesity and insulin resistance due to the decline of estrogen, but genistein, a phytoestrogen, may reduce the risks of these diet-related diseases. In this study, we hypothesized that supplemental genistein has beneficial effects on insulin resistance in an ovariectomized rat model by modulating lipid metabolism. Three weeks after a sham surgery (sham) or an ovariectomy (OVX), ovariectomized Sprague-Dawley rats were placed on a diet containing 0 (OVX group) or 0.1% genistein for 4 weeks. The sham rats were fed a high-fat diet containing 0% genistein and served as the control group (sham group). The ovariectomized rats showed increases in body weight and insulin resistance index, but genistein reduced insulin resistance index and the activity of hepatic fatty acid synthetase. Genistein was also associated with increased activity of succinate dehydrogenase and carnitine palmitoyltransferase and the rate of β-oxidation in the fat tissue of rats. The ovariectomized rats given genistein had smaller-sized adipocytes. Using gene-set enrichment analysis (GSEA) of microarray data, we found that a number of gene sets of fatty acid metabolism, insulin resistance, and oxidative stress were differentially expressed by OVX and reversed by genistein. This systemic approach of GSEA enables the identification of such consensus between the gene expression changes and phenotypic changes caused by OVX and genistein supplementation. Genistein treatment could help reduce insulin resistance through the amelioration of OVX-induced metabolic dysfunction, and the GSEA approach may be useful in proposing putative targets related to insulin resistance.

Topics: Adipose Tissue; Animals; Carnitine O-Palmitoyltransferase; Diet, High-Fat; Dietary Supplements; Disease Models, Animal; Fatty Acid Synthases; Female; Gene Expression; Genistein; Glycine max; Insulin Resistance; Lipid Metabolism; Liver; Microarray Analysis; Obesity; Ovariectomy; Oxidation-Reduction; Oxidative Stress; Phytoestrogens; Phytotherapy; Plant Extracts; Postmenopause; Rats; Rats, Sprague-Dawley; Succinate Dehydrogenase; Weight Gain

Deduce whether the isoflavone genistein blunts the effect of ischaemia to the retina.. Ischaemia was induced in rats by raising the intraocular pressure (120 mm Hg) for 50 min. Genistein (10 mg/kg) was injected intraperitoneally 1 h before and after ischaemia. Seven days after ischaemia, the level of mRNAs for neurofilament light (NF-L), caspase 3, caspase 8, glial fibrillary acidic protein (GFAP), poly-ADP ribose polymerase (PARP), Thy-1 and proteins (GFAP, NF-L, PARP) in whole retinas were determined. NF-L and tubulin proteins in optic nerves were also determined. Retinas were also processed for the localization of choline acetyltransferase (ChAT) and GFAP immunoreactivities.. Ischaemia caused a significant reduction in ganglion cell proteins in the optic nerve (NF-L and tubulin) and retina (NF-L). Retinal Thy-1 (mRNA and protein) and NF-L (mRNA) were also reduced while mRNAs of caspase 3, caspase 8, PARP and GFAP (also protein) were increased. Changes in the mRNAs and proteins induced by ischaemia were significantly blunted by genistein with the exception of the increase in GFAP and PARP protein/mRNA levels. Ischaemia-induced changes in the localization of ChAT were also clearly attenuated by genistein treatment.. Genistein blunts most of the damaging effects caused to the retina by ischaemia.

Topics: Animals; Caspase 3; Caspase 8; Cyclophilins; Disease Models, Animal; Female; Fluorescent Antibody Technique, Indirect; Genistein; Glial Fibrillary Acidic Protein; Injections, Intraperitoneal; Intraocular Pressure; Neurofilament Proteins; Ocular Hypertension; Phytoestrogens; Poly Adenosine Diphosphate Ribose; Proteins; Rats; Rats, Wistar; Reperfusion Injury; Retinal Diseases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thy-1 Antigens

Alzheimer's disease (AD) is a debilitating neurodegenerative disorder characterized by increased β-amyloid (Aβ) deposition and neuronal dysfunction leading to impaired learning and recall. Ageing, heredity, and induced oxidative stress are among proposed risk factors. The increased frequency of the disease in women also suggests a role for estrogen in development of AD. In the present study, effects of the phytoestrogen genistein (10mg/kg) on learning and memory impairments was assessed in intrahippocampal Aβ(1-40)-injected rats. The estrogen receptor antagonist fulvestrant was injected intracerebroventricularly in a group of Aβ-lesioned rats. The Aβ-injected animals exhibited the following: lower spontaneous alternation score in Y-maze tasks, impaired retention and recall capability in the passive avoidance test, and fewer correct choices and more errors in the RAM task. Genistein, but not genistein and fulvestrant, significantly improved most of these parameters. Measurements of oxidative stress markers in hippocampal tissue of Aβ-injected rats showed an elevation of malondialdehyde (MDA) and nitrite content, and a reduction of superoxide dismutase (SOD) activity. Genistein significantly attenuated the increased MDA content but did not affect the nitrite content or SOD activity. These results indicate that genistein pretreatment ameliorates Aβ-induced impairment of short-term spatial memory in rats through an estrogenic pathway and by inducing attenuation of oxidative stress.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Analysis of Variance; Animals; Avoidance Learning; Disease Models, Animal; Estradiol; Estrogen Antagonists; Fulvestrant; Genistein; Hippocampus; Infusions, Intraventricular; Male; Malondialdehyde; Maze Learning; Memory, Short-Term; Mental Recall; Microinjections; Neuroprotective Agents; Oxidative Stress; Peptide Fragments; Phytoestrogens; Random Allocation; Rats; Rats, Wistar; Retention, Psychology; Statistics, Nonparametric; Thiobarbituric Acid Reactive Substances

Chronic intermittent hypoxia (CIH) is a frequent feature of obstructive sleep apnea/hypopnea syndrome (OSAHS), and it may alter upper airway muscle endurance. We have previously reported the positive effects of estrogen on genioglossus fatigue resistance in rats. Our present study was designed to evaluate the effects of two phytoestrogens - genistein and coumestrol - on genioglossus contractile function and estrogen receptor (ER) expression in female rats exposed to CIH. Eight-wk-old female rats were ovariectomized and exposed to CIH for 5 wk. Genistein and coumestrol, respectively, were administered by intraperitoneal injection, at a dose of 2.5 mg kg(-1) d(-1), during the last 4 d of exposure to CIH. The contractile properties of the genioglossus were measured. Real-time RT-PCR and western blotting were performed to determine the expression of ERs in the genioglossus. Phytoestrogens were found to significantly increase genioglossus fatigue resistance, the effect of genistein being more powerful than that of coumestrol. However, higher levels of ER mRNA and protein were detected in the coumestrol group than in the genistein group. We conclude that phytoestrogens, especially genistein, could improve the endurance of the genioglossus muscle in ovariectomized rats exposed to CIH, and this effect is, in part, not related to its estrogenic action.

Topics: Analysis of Variance; Animals; Chronic Disease; Coumestrol; Disease Models, Animal; Female; Genistein; Hypoxia; Muscle Contraction; Muscle, Skeletal; Phytoestrogens; Rats; Rats, Sprague-Dawley; Receptors, Estrogen; RNA, Messenger; Sleep Apnea, Obstructive; Tongue

Soy-derived isoflavones potentially protect against obesity and depression. In five different studies we examined the influence of soy-containing diets or equol injections on depression, serotonin levels, body weight gain (BW) and white adipose tissue (WAT) deposition in female Long-Evans rats at various stages of life [rats were intact, ovariectomized or experienced natural ovarian failure (NOF)].. In general, animals fed a soy-rich diet (Phyto-600) and/or administered equol (@ 5 mg/kg/day) displayed significant decreases in BW and WAT compared to a low-soy diet. When equol was injected alone (5 mg/kg/day), experiments 1, 4, and 5 demonstrated that body weight was significantly decreased. Equol has body weight control effects in females that are dependent on ovarian status and/or age of diet initiation. Experiments 1-4 all displayed no significant differences in depressive-related behavior as measured by the Prosolt forced swim test (PFST) when soy-rich (Phyto-600) or low-soy diets (Phyto-low) or equol treatments (5 mg/kg/day) were tested in female rats at various ages or hormonal status. Results of all the experiments are not presented here due to space limitations, but data from experiment 5 are presented. From conception female rats were exposed to either: a) a soy-rich (Phyto-600) or b) low-soy diet (Phyto-low). After 290 days all rats experienced NOF. At 330 days-old the animals were examined in the Porsolt forced swim test (PFST). One month later a second PFST was performed [after Phyto-low fed animals were injected with equol (5 mg/kg/day) for one week prior to the second PFST]. At the first PFST, serotonin and mobility levels were significantly decreased in the Phyto-low fed animals compared to animals that consumed the Phyto-600 diet. After equol injections at the second PFST, mobility and serotonin levels significantly increased in aged NOF rats fed the Phyto-low diet (to levels comparable to Phyto-600 fed animals).. Consumption of dietary isoflavones or equol exposure in rats has body weight controlling effects and equol specifically may have antidepressant potential dependent upon diet initiation and/or dosage of treatments. The current study demonstrates that equol is able to decrease body weight, abdominal WAT, and depressive-related behavior. While other factors and mechanisms may play a role, in part, the present results provide a greater understanding of how isoflavonoid molecules modulate the brain's influence on behavior.

Topics: Age Factors; Aging; Animals; Body Weight; Depression; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Equol; Female; Glycine max; Hormones; Isoflavones; Male; Obesity; Organ Size; Ovariectomy; Phytoestrogens; Rats; Serotonin; Soybean Proteins; Statistics, Nonparametric; Swimming

Flaxseed, rich in the phytoestrogen lignan secoisolariciresinol diglycoside (SDG), provides protection against bone loss at the lumbar vertebrae primarily when combined with low-dose estrogen therapy in the ovariectomized rat model of postmenopausal osteoporosis. Whether SDG metabolites are accessible to skeletal tissue, and thus have the potential to interact with low-dose estrogen therapy to exert direct local action on bone metabolism, is unknown. The objective of this study was to determine whether metabolites of SDG are accessible to the skeleton of ovariectomized rats and to compare the distribution of SDG metabolites in skeletal tissue with that in other tissues. Rats were fed a 10% flaxseed diet and gavaged daily with tritium-labeled SDG (7.4 kBq/g of body weight) in deionized water (500 μL) (n=3) or deionized water alone (n=3) for 7 days, after which tissues were collected for liquid scintillation counting. Radioactivity was detected in similar concentrations in the lumbar vertebrae, femurs, and tibias. Compared with non-skeletal tissues, total radioactivity in the skeleton was significantly lower than in the liver, heart, kidney, thymus, and brain (P < .001). There were no significant differences in levels of radioactivity between skeletal tissue versus the spleen, lung, bladder, uterus, vagina, and mammary gland. In conclusion, SDG metabolites are accessible to skeletal tissue of ovariectomized rats. Thus, it is biologically plausible that SDG metabolites may play a direct role in the protective effects of flaxseed combined with low-dose estrogen therapy against the loss of bone mass and bone strength in the ovariectomized rat model of postmenopausal osteoporosis.

Topics: Animals; Butylene Glycols; Disease Models, Animal; Female; Femur; Flax; Lignans; Lumbar Vertebrae; Osteoporosis; Ovariectomy; Phytoestrogens; Rats; Rats, Sprague-Dawley; Tibia

As an alternative to estrogen therapy, the efficacy of an estrogen receptor β-selective phytoestrogenic (phyto-β-SERM) formulation to regulate climacteric symptoms and decline in brain responses associated with ovarian hormone loss in menopause was assessed.. A phyto-β-SERM formulation-containing diet was compared with a commercial soy extract diet and a phytoestrogen-free base/control diet in an ovariectomized (OVX) mouse model of human menopause. Two treatment studies were conducted: (1) a 2-month study assessed the effects of experimental diets on tail skin temperature as a model of menopausal hot flashes, and (2) a 9-month study assessed the long-term impact of the diets on overall health, hair thinning/loss, spatial working memory, and associated protein expression in the hippocampus.. The phyto-β-SERM diet prevented OVX-induced menopause-like changes including the rise in skin temperature, hair thinning/loss, deficit in spatial memory function, and reversed OVX-induced decline in the expression of hippocampal proteins involved in neural plasticity and β-amyloid degradation/clearance. The soy extract diet had no effect or exacerbated OVX-induced changes.. Overall, the phyto-β-SERM diet induced physical and neurological responses comparable with ovary-intact mice, suggesting the therapeutic potential of the phyto-β-SERM formulation for the prevention/alleviation of climacteric symptoms and decline in brain responses induced by ovarian hormone loss, which provides the basis for further work in postmenopausal women.

Topics: Amyloid beta-Peptides; Animals; Disease Models, Animal; Estrogen Receptor beta; Female; Hair; Hippocampus; Hot Flashes; Humans; Memory Disorders; Memory, Short-Term; Menopause; Mice; Mice, Inbred C57BL; Neuronal Plasticity; Phytoestrogens

This experiment established the ovariectomized (OVX) rat model of postmenopausal osteoporosis and examined the effect of the oral administration of different dosages of dioscorea, red mold dioscorea (RMD), and soy isoflavones on bone mineral density (BMD). Three months after osteoporosis had been induced and 4 weeks after feeding had begun, the tibia and femur BMD of OVX rats administered RMD showed significant increases compared with that of all other groups of OVX rats. Closer examination using microcomputed tomography also revealed that the RMD-administered rats had denser trabecular bone volume and a higher trabecular number compared to all other rat groups. Reconstructed 3D imaging indicated increases in cancellous bone mineral content, cancellous bone mineral density, and cortical bone mineral content of the proximal tibia in OVX rats. These findings indicate that administration of monacolin K and phytoestrogen diosgenin could prevent bone loss induced by estrogen deficiency.

Topics: Animals; Bone Density; Dioscorea; Diosgenin; Disease Models, Animal; Female; Fermentation; Flavins; Glycine max; Heterocyclic Compounds, 3-Ring; Isoflavones; Lovastatin; Monascus; Osteoporosis; Ovariectomy; Phytoestrogens; Plant Roots; Rats; Rats, Sprague-Dawley

Preimplantation embryo loss during oviduct transit has been observed in adult mice after a 5-day neonatal exposure to the phytoestrogen genistein (Gen; 50 mg/kg/day).. We investigated the mechanisms underlying the contribution of the oviduct to infertility.. Female mice were treated on postnatal days 1-5 with corn oil or Gen (50 mg/kg/day). We compared morphology, gene expression, and protein expression in different regions of the reproductive tracts of Gen-treated mice with those of control littermates at several time points.. Neonatal Gen treatment resulted in substantial changes in expression of genes that modulate neonatal oviduct morphogenesis, including Hoxa (homeobox A cluster), Wnt (wingless-related MMTV integration site), and hedgehog signaling genes. An estrogen receptor antagonist blocked these effects, indicating that they were induced by the estrogenic activity of Gen. Oviducts of adults treated neonatally with Gen had abnormal morphology and were stably "posteriorized," as indicated by altered Hoxa gene patterning during the time of treatment and dramatic, permanent up-regulation of homeobox genes (e.g., Pitx1, Six1) normally expressed only in the cervix and vagina.. Neonatal exposure to estrogenic environmental chemicals permanently disrupts oviduct morphogenesis and adult gene expression patterns, and these changes likely contribute to the infertility phenotype.

Topics: Animals; Animals, Newborn; Disease Models, Animal; Female; Gene Expression Regulation; Genistein; Immunoblotting; Infertility; Mice; Microarray Analysis; Morphogenesis; Oviducts; Phytoestrogens; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors

Daidzein is a potential natural alternative to estradiol during therapy of some malignancies in men. Besides weak inhibition of tyrosine kinase activity, daidzein has a sizeable inhibitory effect on calcium channels. The aim of this study was to examine the effects of daidzein on the immunohistomorphometric features of pituitary adrenocorticotropes (ACTH cells) and circulating levels of ACTH and corticosterone, in comparison with estradiol, in an animal model of the andropause. Sixteen-month-old Wistar rats were divided into sham operated (SO), orchidectomized (Orx), estradiol treated orchidectomized (Orx+E) and daidzein treated orchidectomized (Orx+D) groups. Estradiol (0.625 mg/kg/day) and daidzein (30 mg/kg/day) were administered subcutaneously for three weeks, while the SO and Orx groups received the vehicle alone. ACTH cells were identified by the peroxidase-antiperoxidase (PAP) immunohistochemical procedure. Peripheral circulating concentrations of ACTH and corticosterone were measured by immunoassay. Orchidectomy reduced (p<0.05) the cell volume and volume density of adrenocorticotropes by 11% and 16%, respectively, in comparison to SO rats. In Orx+E rats, the volume density of ACTH cells decreased (p<0.05) by 25%, but the circulating level of ACTH increased (p<0.05) by 29%, compared to Orx rats. Daidzein treatment significantly decreased (p<0.05): volume density of ACTH cells, circulating ACTH and corticosterone by 24%, 48% and 33%, respectively, compared to the Orx group. In conclusion, this study revealed that daidzein negatively modulated the immunohistomorphometric features of ACTH cells and, unlike estradiol, decreased ACTH and corticosterone secretion, in an animal model of the andropause.

Topics: Andropause; Animals; Corticosterone; Corticotrophs; Disease Models, Animal; Estradiol; Estrogens; Immunohistochemistry; Isoflavones; Male; Orchiectomy; Phytoestrogens; Rats; Rats, Wistar

Mucopolysaccharidoses (MPS) are lysosomal storage disorders resulting from a deficit of specific lysosomal enzymes catalysing glycosaminoglycan (GAG) degradation. The typical pathology involves most of the organ systems, including the brain, in its severe forms. The soy isoflavone genistein has recently attracted considerable attention as it can reduce GAG synthesis in vitro. Furthermore, genistein is able to cross the blood-brain barrier in the rat. The present study was undertaken to assess the ability of genistein to reduce urinary and tissue GAG levels in vivo.. We used mice with genetic deletion of iduronate-2-sulphatase (one of the GAG catabolizing enzymes) which provide a model of MPS type II. Two doses of genistein, 5 or 25 mg.kg(-1).day(-1), were given, in the diet for 10 or 20 weeks. Urinary and tissue GAG content was evaluated by biochemical and histochemical procedures.. Urinary GAG levels were reduced after 10 weeks' treatment with genistein at either 5 or 25 mg.kg(-1).day(-1). In tissue samples from liver, spleen, kidney and heart, a reduction in GAG content was observed with both dosages, after 10 weeks' treatment. Decreased GAG deposits in brain were observed after genistein treatment in some animals.. There was decreased GAG storage in the MPSII mouse model following genistein administration. Our results would support the use of this plant-derived isoflavone in a combined therapeutic protocol for treatment of MPS.

Topics: Animals; Blood-Brain Barrier; Brain; Disease Models, Animal; Dose-Response Relationship, Drug; Genistein; Glycosaminoglycans; Iduronate Sulfatase; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mucopolysaccharidosis II; Phytoestrogens

Genistein has been implicated in the beneficial effects of soy on human health, particularly in the context of ageing. In post-menopausal women reduced systemic estrogen leads to a range of age-associated pathologies, including delayed cutaneous wound healing. We have previously shown that this can be reversed by estrogen replacement. However, the effect of genistein on the skin is poorly understood and crucially the influence of genistein on wound healing has not been assessed. 10-week-old ovariectomised mice were systemically treated with 17beta-estradiol or genistein. Genistein substantially accelerated wound repair, associated with a dampened inflammatory response. Unexpectedly, co-treatment with the ER antagonist ICI had little impact on the anti-inflammatory, healing promoting effects of genistein. Thus genistein's actions are only partially mediated via classical estrogen receptor-dependent signalling pathways. Indeed, we report that alternative (cell-type specific) signalling mechanisms are activated in the skin in response to genistein treatment.

Topics: Animals; Anti-Inflammatory Agents; Base Sequence; Cells, Cultured; Disease Models, Animal; Estrogens; Female; Gene Expression Regulation; Genistein; Immunohistochemistry; Macrophages; Mice; Mice, Inbred C57BL; Molecular Sequence Data; Ovariectomy; Phytoestrogens; Polymerase Chain Reaction; Receptors, Estrogen; Wound Healing

Plant-derived estrogen-like compounds such as isoflavones (IF) especially daidzein and genistein are said to be preserving the bone in the osteoporotic conditions. However, it is not known whether a combination of IF and calcium (Ca) supplementation attenuates losses in bone mass and prevents the loss of vitamin D (VD). The present study addresses the role of phytoestrogens (PE) and Ca supplementation in low Ca and low VD diet induced osteoporosis (OSP). Cowpea (CP) which has high amount of the IF was selected to study its effect on diet induced osteoporotic conditions. Female weanling WNIN rats (total of 68) were divided into five groups and fed for five weeks on semisynthetic diet with low Ca (0.15%) and low VD (0.1IU/day/rat) in combination with low (10 mg/kg) or high (25 mg/kg) concentrations of PEs derived from CPIF. The study groups are: (I) normal Ca(0.47%) and normal VD (25IU/day/rat), (II) low Ca+low VD, (III) low Ca+low VD+low CPIF (10 mg/kg diet), (IV) low Ca+low VD+high CPIF (25 mg/kg diet) and (V) low Ca+low VD+17-(-estradiol (3.2 mg/kg diet). After the development of OSP the group II was subgrouped into: (SG I) continued on low Ca+VD, (SG II) low CPIF, (SG III) high CPIF, (SG IV) 17-beta-estradiol and (SG V) normal Ca and VD. Serum 25-VD levels were in the range of 14-38 ng/ml in groups I, III, IV and V, where as the values were very low in the group II (5.8 ng/ml). These were partially reversed upon supplementation of CPIF. The results correlated with altered Ca levels, body weight, bone mineral density and content and other related biochemical parameters. The paper further explains the possibility of protective and therapeutic role of VD in the presence of CPIF in osteoporotic health manifestations.

Topics: Animals; Bone Density; Calcium; Cholecalciferol; Dietary Supplements; Disease Models, Animal; Female; Genistein; Isoflavones; Osteoporosis; Phosphorus; Phytoestrogens; Plants; Rats; Vitamin D

Oestrogen loss at menopause is frequently related to poor wound healing. Genistein has been tested in anti-ageing cosmetic preparations with interesting results on skin health. Here, we investigated the effects of the genistein aglycones, given systemically, in an incisional model of wound healing, compared to systemic oestradiol and raloxifene.. Six months after ovariectomy (OVX), rats were randomly assigned to groups of 12 animals each and treated daily with genistein aglycone (1 and 10 mg kg(-1) s.c.), raloxifene hydrochloride (0.05 and 0.5 mg kg(-1) s.c.) or 17-alpha-ethinyl oestradiol (0.003 and 0.03 mg kg(-1) s.c.) for 12 weeks. Untreated OVX and sham OVX rats were used as controls. Then, 14 or 7 days before the end of the experiment, an incisional wound healing procedure was performed and skin specimens were collected to evaluate molecular, histological and functional measurements.. Seven and fourteen days after wounding, samples from OVX rats showed a decrease in transforming growth factor-beta1, tissue transglutaminase 2 and vascular endothelial growth factor compared to samples from sham OVX rats. Oestradiol, raloxifene and genistein all significantly modified this decrease, but the lowest genistein dose exerted a greater effect than the other treatments. Moreover, the lowest dose of genistein was the most effective in improving skin healing and wound tensile strength.. Genistein aglycone might be an alternative therapy for the management of skin wound healing.

Topics: Animals; Dermatologic Surgical Procedures; Disease Models, Animal; Dose-Response Relationship, Drug; Ethinyl Estradiol; Female; Genistein; GTP-Binding Proteins; Ovariectomy; Phytoestrogens; Protein Glutamine gamma Glutamyltransferase 2; Raloxifene Hydrochloride; Random Allocation; Rats; Rats, Sprague-Dawley; Skin; Tensile Strength; Transforming Growth Factor beta1; Transglutaminases; Vascular Endothelial Growth Factor A; Wound Healing

The role of soy in reducing breast cancer risk has been suggested to be associated with early exposure to isoflavones, which alter mammary gland morphology. The objective of the study was to determine the effect of dietary exposure to the enantiomers of a key soy isoflavone metabolite, equol, on mammary gland development and later chemoprotection using the DMBA-induced animal model of breast cancer. Animals were exposed to S-(-)equol or R-(+)equol (250 mg/kg diet) during the neonatal (0-21 days) or prepubertal (21-35 days) periods only. Histological evaluation of the mammary glands showed that both enantiomers fed neonatally via the dam led to significant precocial mammary gland differentiation. By day 50, early S-(-)equol or R-(+)equol exposure resulted in a decrease in immature terminal end structures and an increase in mature lobules, suggesting an early 'imprinting' effect. Despite these morphological changes to the mammary gland, neonatal and prepubertal exposure to equol had no long-term chemoprevention against mammary tumors induced by DMBA, although for R-(+)equol there was a trend to delaying tumor formation. In summary, early exposure to equol was not chemopreventive, but neither did it increase tumor formation in response to DMBA, suggesting exposure in early life does not influence breast cancer risk.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Animals, Newborn; Body Weight; Carcinogens; Disease Models, Animal; Equol; Female; Genistein; Isoflavones; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Organ Size; Phytoestrogens; Rats; Rats, Sprague-Dawley; Stereoisomerism; Time Factors

To observe the estrogenic effect of formononetin and its effect on the expressions of atrial estrogen receptor subtypes alpha and beta (ERalpha and ERbeta).. 50 femal rats were randomly divided into five groups: sham group, model group, nilestriol group, formononetin groups of low and high dose. Rats in sham group were cut a piece of fat before closing the abdomen, the others were ovariectomized. Vaginal exfoliated cell were observed from the fifth day to the tenth after operation to test if the model is successful. The sham and model group were given nomal saline in 10 mL/kg by gavage, the remaining three groups were given nilestriol 2.5 mg/(kg x w), low [20 mg/(kg x d) land high dose [100 mg/(kg x d)) of formononetin by gavage respectively. In the 8th week, vaginal exfoliated cell were observed, then decapitated the rats, removed the uterus, weighed and take wright staining microscopy. The relative expressions of ERalpha and ERbeta of right atrium were detected by RT-PCR.. The vaginal cells exhibit a change of estrus after had been fed with high dose of formononetin after 8 weeks. Formononetin increase the uterus coefficient and the expression of atrial ERbeta (P < 0.01), but it dose not have any effect on the expression of ERalpha (P > 0.05).. Formononetin have estrogenic effect in ovariectomized rats, and it can markedly upregulate the expression of rats' atrial ERbeta.

Topics: Administration, Oral; Animals; Disease Models, Animal; Estriol; Estrogen Receptor alpha; Estrogen Receptor beta; Female; Heart Atria; Isoflavones; Ovariectomy; Phytoestrogens; Quinestrol; Random Allocation; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Trifolium; Uterus

A large body of experimental evidence supports a role for oxidative stress as a mediator of nerve cell death in Parkinson's disease (PD). Phytoestrogens such as genistein have been reported to prevent neuronal degeneration caused by increased oxidative burden, therefore, this study examined whether genistein administration at a high dose would attenuate behavioral and structural abnormalities in an experimental model of PD in rat. For this purpose, unilateral intrastriatal 6-hydroxydopamine (6-OHDA, 12.5 microg/5 microL of saline-ascorbate)-lesioned rats were intraperitoneally pretreated with a single and high dose of genistein (10 mg/kg) 1 h before surgery. Apomorphine-induced rotations and the number of Nissl-stained neurons in the substantia nigra pars compacta (SNC) were counted after 2 weeks. Genistein administration could attenuate the rotational behavior in lesioned rats and protect the neurons of SNC against 6-OHDA toxicity. Genistein administration has a protective effect against 6-OHDA toxicity.

Topics: Animals; Antiparkinson Agents; Behavior, Animal; Disease Models, Animal; Genistein; Male; Motor Activity; Oxidative Stress; Oxidopamine; Parkinson Disease, Secondary; Phytoestrogens; Rats; Rats, Sprague-Dawley; Substantia Nigra

Female mice treated neonatally with the phytoestrogen genistein (50 mg/kg/day) have multioocyte follicles, lack regular estrous cyclicity, and are infertile even after superovulation. To determine the cause of their infertility, we examined oocyte developmental competence and timing of embryo loss. Eggs obtained by superovulation of genistein-treated or control females were equally capable of being fertilized in vitro and cultured to the blastocyst stage. However, if eggs were fertilized in vivo, retrieved at the pronucleus stage, and cultured, there was a significant reduction in the percentage of embryos from genistein-treated females reaching the blastocyst stage. When these blastocysts were transferred to pseudopregnant recipients, the number of live pups produced was similar to that in controls. Preimplantation embryo development in vivo was examined by flushing embryos from the oviduct and/or uterus. Similar numbers of one-cell and two-cell embryos were obtained from genistein-treated and control females. However, significantly fewer embryos (<50%) were obtained from genistein-treated females on postcoital Days 3 and 4. To determine if neonatal genistein treatment altered the ability of the uterus to support implantation, blastocysts from control donors were transferred to control and genistein-treated pseudopregnant recipients. These experiments demonstrated that genistein-treated females are not capable of supporting normal implantation of control embryos. Taken together, these results suggest that oocytes from mice treated neonatally with genistein are developmentally competent; however, the oviductal environment and the uterus have abnormalities that contribute to the observed reproductive failure.

Topics: Animals; Cells, Cultured; Disease Models, Animal; Embryo Implantation; Embryo Transfer; Embryonic Development; Female; Fertilization; Genistein; Infertility, Female; Mice; Mice, Inbred ICR; Oocytes; Ovary; Oviducts; Phytoestrogens; Pregnancy; Prenatal Exposure Delayed Effects; Uterus

Administration of the isoflavone genistein (GEN) has been described to result in bone protection but also to induce uterotrophic responses. To compare bone protective effects of GEN with an isoflavone-rich diet (IRD) and to further elucidate molecular mechanisms involved in bone-protection, ovariectomized rats (OVX) received either a diet low in isoflavone content (IDD) enriched with GEN (42 mg kg(-1)b.wtd(-1)) (GEN(d)), an IRD (14 mg kg(-1)b.wtd(-1) GEN, 14 mg kg(-1)b.wtd(-1) daidzein) or were treated subcutaneously (s.c.) with GEN (10 mg kg(-1)b.wtd(-1)) (GEN(sc)) for 12 weeks. Intact (SHAM), vehicle treated OVX animals and those substituted with 17beta-estradiol (2microg kg(-1)b.wtd(-1)) (E(2)), served as controls. OVX-induced bone loss could be antagonized in E(2), GEN(sc), GEN(d) and IRD groups. Uterine wet weight (UWW) was only stimulated in E(2) and GEN(sc) animals. Serum biomarkers of bone-formation (osteocalcin, osteopontin) and bone-resorption (telopeptides of collagen type I, pyridinoline cross-links) were elevated in OVX compared to SHAM and E(2) animals. Feeding IRD stimulated bone-formation and inhibited bone-resorption, whereas s.c. or dietary administration of GEN only resulted in a stimulation of bone-formation. The results of the present study indicate that in contrast to s.c. administration, dietary intake of GEN resulted in bone protection without stimulation of UWW. Dietary intake of isoflavones by an IRD also did not result in a stimulation of UWW, yet IRD appeared to be more effective in bone protection than administration of pure GEN.

Topics: Animals; Bone Density; Bone Density Conservation Agents; Bone Resorption; Collagen Type I; Diet; Disease Models, Animal; Estradiol; Female; Genistein; Injections, Subcutaneous; Organ Size; Osteocalcin; Osteogenesis; Osteopontin; Osteoporosis; Ovariectomy; Peptides; Phytoestrogens; Rats; Rats, Wistar; Tibia; Uterus

Healing of predominantly metaphyseal fractures in postmenopausal osteoporosis is delayed and comparatively poor. Hormone replacement therapy could improve fracture healing, but, because of its potential side effects, natural alternatives are more appealing. The aim of this study was to determine if the soy metabolite equol and the native isoflavone genistein, in comparison to 17beta-estradiol, improve metaphyseal fracture healing in ovariectomy-induced osteoporotic bone of the rat. Forty-eight 12-week-old female rats developed severe osteoporosis ten weeks after ovariectomy. After metaphyseal tibial osteotomy and standardized stable internal fixation, changes in callus morphology were evaluated biomechanically, qualitatively and quantitatively in fluorochrome-labeled histological sections and microradiographs in ovariectomized rats (C) and under standardized 17beta-estradiol (E), equol (EQ) and genistein (G) supplemented rats over a period of five weeks. Estrogen and equol were able to improve the elasticity of callus formation significantly in postmenopausal osteoporotic bone (stiffness of C: 121.40 +/- 47.08 N/mm, E: 147.90 +/- 39.38 N/mm, EQ: 167.8 +/- 59.90 N/mm). The effects of estrogen were more anabolic than those of equol and were visible in changes to the trabecular bone (N.Nd of E: 6.47 +/- 7.68, EQ: 4.25 +/- 3.96). However, in terms of the whole body, equol seemed to induce less of an adverse reaction than estrogen (body weight of C: 342.20 +/- 19.91 g, E: 280.25 +/- 12.05 g, EQ: 308.75 +/- 24.28 g). Genistein as an osteoclast inhibitor influenced callus stiffness (G: 144.50 +/- 61.52 N/mm) and negatively impacted trabecular structure (N.Nd of G: 0.59 +/- 1.01) in severely osteoporotic bones. Estrogen and equol were able to improve fracture healing in ovariectomy-induced osteoporotic bones, and the extent of callus formation played only a minor role. Genistein rather negatively influenced fracture healing. The metaphyseal osteotomy model in ovariectomized rats allows an accurate study of the therapeutic effects of antiosteoporotic substances on the fracture healing process.

Topics: Animals; Bony Callus; Disease Models, Animal; Elasticity; Equol; Estradiol; Estrogens; Female; Fracture Healing; Genistein; Glycine max; Isoflavones; Osteoclasts; Osteoporosis; Ovariectomy; Phytoestrogens; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley; Tibia

Some degenerative diseases of the nervous system have been linked to hormonal imbalance in postmenopausal women. It is argued that young coconut juice (YCJ) could have some estrogen-like characteristics, but this is still debatable. Our aim was to investigate this argument, and to examine whether YCJ has any neuroprotective effects.. Four groups of female rats (10 in each group) were included in this study. These included sham-operated, overiectomized (ovx), ovx and receiving estradiol benzoate (EB) injections intraperitoneally, and ovx and receiving YCJ orally. At the end of the five-week study, the rats were sacrificed, and their serum estradiol (E2) level was measured by chemiluminescent immunoassay. Moreover, the rat brains were excised, and the cortical pyramidal neurons were examined using markers of neuronal cell death, namely anti-neurofilament (NF200) and anti-parvalbumin (PV) antibodies.. Our results showed that the rat group which received YCJ had its serum E2 level significantly (P<0.05) higher than the ovx group which did not receive any treatment, and the sham-operated group. A similar trend was observed with the group which received EB injections, but no significant difference was present when the latter was compared with the sham-operated group. In addition, a significant reduction in neuronal cell death was observed in the YCJ-treated group, as compared to the ovx group which did not receive any treatment. This was indicated by the significantly (P<0.05) higher number of neurons which were immunopositive for NF200 and PV. Interestingly, the number of these neurons was also significantly (P<0.05) higher in the YCJ group, as compared to the EB group.. This study confirms the argument that YCJ has estrogen-like characteristics, and it also adds more evidence to the observation that hormonal imbalance could induce some brain pathologies in females.

Topics: Animals; Brain; Cell Death; Cocos; Disease Models, Animal; Estradiol; Female; Fruit; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Neurofilament Proteins; Neurons; Ovariectomy; Parvalbumins; Phytoestrogens; Phytotherapy; Plant Extracts; Postmenopause; Rats; Rats, Wistar

Genistein, a major phytoestrogen of soy, is considered a potential drug for prevention and treatment of postmenopausal osteoporosis. The aim of the present study was to compare the effects of genistein, estradiol and raloxifene on the skeletal system in vivo and in vitro. Genistein (5 mg/kg), estradiol (0.1 mg/kg) or raloxifene hydrochloride (5 mg/kg) were administered daily by a stomach tube to mature ovariectomized Wistar rats for 4 weeks. Bone mass, mineral and calcium content, macrometric parameters and mechanical properties were examined. Also the effects of genistein, estradiol and raloxifene (10(-9)-10(-7) M) on the formation of osteoclasts from neonatal mouse bone marrow cells and the activity of osteoblasts isolated from neonatal mouse calvariae were compared. In vivo, estrogen deficiency resulted in the impairment of bone mineralization and bone mechanical properties. Raloxifene but not estradiol or genistein improved bone mineralization. Estradiol fully normalized the bone mechanical properties, whereas genistein augmented the deleterious effect of estrogen-deficiency on bone strength. In vitro, genistein, estradiol and raloxifene inhibited osteoclast formation from mouse bone marrow cells, decreasing the ratio of RANKL mRNA to osteoprotegerin mRNA expression in osteoblasts. Genistein, but not estradiol or raloxifene, decreased the ratio of alkaline phosphatase mRNA to ectonucleotide pyrophosphatase phosphodiesterase 1 mRNA expression in osteoblasts. This difference may explain the lack of genistein effect on bone mineralization observed in ovariectomized rats in the in vivo study. Concluding, our experiments demonstrated profound differences between the activities of genistein, estradiol and raloxifene towards the osseous tissue in experimental conditions.

Topics: Animals; Body Weight; Bone and Bones; Bone Density; Bone Marrow Cells; Bone Resorption; Calcification, Physiologic; Calcium; Cell Count; Cells, Cultured; Disease Models, Animal; Drug Combinations; Estradiol; Estriol; Female; Gene Expression; Genistein; Humans; Mice; Mice, Inbred BALB C; Organ Size; Osteoblasts; Osteoclasts; Osteoporosis, Postmenopausal; Ovariectomy; Phytoestrogens; Raloxifene Hydrochloride; Rats; Rats, Wistar; RNA, Messenger; Selective Estrogen Receptor Modulators; Thymus Gland; Uterus

Plant-derived phytoestrogens have bone protective effects, but the molecular mechanism behind these effects remains unclear. This study is aimed at fully characterizing the fracture healing process of formononetin, and investigating the mechanism underlying angiogenesis in calluses of a rat fracture model. Femoral fractures were produced in 2-month-old Sprague-Dawley rats. A 20 microg/kg or 200 microg/kg dose of formononetin was orally administrated once a day during the healing period of 21 days. The results showed that in the early stage of chondrogenesis (days 3), formononetin significantly increased the number of vessels, and expression of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR-2/flk-1) compared with control. However, the larger dose of formononetin had no significant difference on expression of VEGF and VEGFR-2/Flk-1 compared with that of the smaller dose of formononetin. After 7 days of administration, formononetin markedly induced differentiation of mesenchymal stem cells in the fracture site. After 14 days, gene expression of mesenchymal progenitors such as alkaline phosphatase (ALP), osteocalcin (OCN), osteopontin (OPN) and collagen type I (Col I), indicating osteogenic differentiation, was markedly stimulated by formononetin compared with control. These results suggest that formononetin promotes early fracture healing through angiogenesis activation in the early stage of fracture repair, and osteogenesis acceleration in the later stages, and thus may be beneficial for fracture healing.

Topics: Animals; Astragalus propinquus; Bony Callus; Cell Differentiation; Chondrogenesis; Collagen Type I; Disease Models, Animal; Femur; Fractures, Bone; Isoflavones; Male; Mesenchymal Stem Cells; Neovascularization, Physiologic; Osteocalcin; Osteopontin; Phytoestrogens; Phytotherapy; Plant Extracts; Plant Roots; Rats; Rats, Sprague-Dawley; Receptors, Vascular Endothelial Growth Factor; Vascular Endothelial Growth Factor A; Wound Healing

Genistein is a phytoestrogen that is known to have a protective effect on the vascular endothelial wall. However, it exhibits poor bioavailability, which limits the use of genistein to treat cardiovascular and cerebrovascular diseases. A novel genistein derivative, 7-difluoromethyl-5,4'-dimethoxygenistein (dFMGEN), has shown a better protective effect on vascular endothelial damage in vitro than genistein. In this study, we further evaluated therapeutic effects of dFMGEN on the vascular endothelial wall and atherosclerosis in a rabbit model in vivo. There were 5 groups: the GEN group (genistein 5 mg/kg per day), lovastatin group (lovastatin 5 mg/kg per day), dFMGEN group (dFMGEN 5 mg/kg per day), model control group (the same amount of vehicle solvent), and the normal control group; all feedings administered via intragastric administration. We demonstrated that dFMGEN (1) attenuated the development of atherosclerosis, (2) reduced serum total cholesterol and low-density lipoprotein cholesterol concentrations, (3) decreased lipid peroxidation in the rabbit atherosclerosis model, and (4) increased smooth muscle cell and collagen content in atheroma of thoracic aortas. These results provide an experimental foundation for dFMGEN's potential effects in preventing and treating atherosclerosis, acute coronary syndromes, and potentially ischemia-reperfusion injury during acute myocardial infarction and cerebral infarction.

Topics: Animals; Aorta, Abdominal; Aorta, Thoracic; Atherosclerosis; Cholesterol; Collagen; Disease Models, Animal; Endothelium, Vascular; Genistein; Lipid Peroxidation; Lipoproteins, LDL; Male; Phytoestrogens; Rabbits

Cardiac hypertrophy is a common finding in human patients with inborn errors of long-chain fatty acid oxidation. Mice with either very long-chain acyl-coenzyme A dehydrogenase deficiency (VLCAD-/-) or long-chain acyl-coenzyme A dehydrogenase deficiency (LCAD-/-) develop cardiac hypertrophy. Cardiac hypertrophy, initially measured using heart/body weight ratios, was manifested most severely in LCAD-/- male mice. VLCAD-/- mice, as a group, showed a mild increase in normalized cardiac mass (8.8% hypertrophy compared with all wild-type (WT) mice). In contrast, LCAD-/- mice as a group showed more severe cardiac hypertrophy (32.2% increase compared with all WT mice). On the basis of a clear male predilection, we analyzed the role of dietary plant estrogenic compounds commonly found in mouse diets because of soy or alfalfa components providing natural phytoestrogens or isoflavones in cardioprotection of LCAD-/- mice. Male LCAD-/- mice fed an isoflavone-free test diet had more severe cardiac hypertrophy (58.1% hypertrophy compared with WT mice fed the same diet). There were no significant differences in the female groups fed any of the diets. Echocardiography measurement performed on male LCAD-deficient mice fed a standard diet at the age of approximately 3 months confirmed the substantial cardiac hypertrophy in these mice compared with WT controls. Left ventricular (LV) wall thickness of the interventricular septum and posterior wall was remarkably increased in LCAD-/- mice compared with that of WT controls. Accordingly, the calculated LV mass after normalization to body weight was increased by about 40% in the LCAD-/- mice compared with WT mice. In summary, we found that metabolic cardiomyopathy, expressed as hypertrophy, developed in mice because of either VLCAD deficiency or LCAD deficiency; however, LCAD deficiency was the most profound and seemed to be attenuated either by endogenous estrogen (in females) or by phytoestrogens present in the diet as isoflavones (in males).

Topics: Acyl-CoA Dehydrogenase, Long-Chain; Animals; Body Weight; Cardiomegaly; Diet; Disease Models, Animal; Echocardiography; Female; Isoflavones; Male; Mice; Mice, Knockout; Myocardium; Organ Size; Phytoestrogens

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Anticarcinogenic Agents; Apoptosis; Carcinoma; Chemoprevention; Chickens; Disease Models, Animal; Drug Delivery Systems; Fatty Acids, Omega-3; Female; Humans; Mice; Neoplasm Proteins; Ovarian Neoplasms; Phytoestrogens; Progestins; Resveratrol; Retinoids; Stilbenes; Vitamin D

Genistein, a naturally occurring isoflavonic phytoestrogen associated with reduced incidence of heart disease, may be a possible alternative treatment for postmenopausal women with heart disease.. This study examined the effects of genistein on in vitro heart function and ischemic tolerance in ovariectomized (OVX) Sprague-Dawley rats.. To examine the acute effects of genistein on cardiac function, isolated working hearts were perfused under aerobic conditions with increasing concentrations of genistein (10-150 microM). A separate group of OVX rats was used to assess ischemic tolerance: treated rats received genistein (250 mg/kg, dissolved in 200 microL dimethyl sulfoxide [DMSO]) injected once daily for 2 days, and control rats received DMSO only. After treatment, hearts were perfused for 30 minutes under aerobic conditions and then subjected to 20 minutes of global no-flow ischemia by clamping the preload and afterload lines, followed by 30 minutes of reperfusion.. Genistein was associated with improvements in mechanical function in OVX rat hearts (n = 5) with maximum increases in contractility (259 mm Hg/sec above baseline) and cardiac output (7 mL/min above baseline) observed with 30 microM of genistein (both, P < 0.05). Relative to baseline, genistein-treated hearts (n = 5) also had greater ischemic tolerance than did control hearts (n = 6) and significant improvements in mean (SEM) recovery of contractility (to 75.0% [9.7%] of preischemic function; P < 0.05) and cardiac output (to 48.8% [12.3%] of preischemic function; P < 0.05) after reperfusion. These effects occurred without significant changes in myocardial levels of nonprotein thiols or thiobarbituric acid reactive substances, although a reduction in mean glucose transporter protein 4 content (13.2% [2.7%]; P < 0.05) was observed in genistein-treated hearts. No significant changes in blood pressure were observed with genistein.. Despite the lack of significant changes in physical characteristics, 2-day treatment with genistein was associated with significant cardioprotective effects in OVX rats, suggesting a potential therapeutic role in postmenopausal women.

Topics: Animals; Disease Models, Animal; Female; Genistein; In Vitro Techniques; Ischemia; Myocardial Contraction; Ovariectomy; Phytoestrogens; Postmenopause; Rats; Rats, Sprague-Dawley

There is great interest in soy isoflavones as alternatives to endogenous estrogens not only in hormonal pathologies, but also in inflammatory, neurodegenerative diseases, and pain. We investigated the effect of the isoflavone genistein on neuropathic pain. Genistein binds estrogen receptors (ER) with higher affinity for the ERbeta particularly expressed in neuronal and immune cells. Neuropathy was induced in mice by means of chronic sciatic nerve constriction, and the subcutaneous administration of genistein from the third day after the lesion reversed pain hypersensitivity in a time- and dose-dependent manner. This effect may have been due to the activation of classical nuclear receptor and/or anti-oxidant, anti-inflammatory, and immunomodulating properties of genistein. The fact that a specific ERbeta antagonist prevented both its anti-allodynic and anti-hyperalgesic action, whereas a specific ERalpha antagonist was ineffective and a non-selective ER antagonist only reversed the anti-allodynic effect, suggests the involvement of ERbeta. Antioxidant effects are also involved as the anti-nociceptive dose reversed the increase in reactive oxygen species and malondialdehyde in injured paw tissues, and increased the activity of anti-oxidant enzymes. The phytoestrogen had immunomodulatory and anti-inflammatory activities as it reduced peripheral and central nuclear factor-kappaB, nitric oxide system and pro-inflammatory cytokine over-activation. Taken together, our results suggest that genistein could ameliorate painful neuropathy by multiple mechanisms.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Chronic Disease; Disease Models, Animal; Dose-Response Relationship, Drug; Ganglia, Spinal; Genistein; Glycine max; Hyperalgesia; Inflammation; Inflammation Mediators; Ligation; Male; Mice; Mice, Inbred C57BL; Neurons, Afferent; Nociceptors; Oxidative Stress; Peripheral Nervous System Diseases; Phytoestrogens; Plant Extracts; Receptors, Estrogen; Sciatic Neuropathy; Treatment Outcome

Genistein aglycone positively affects bone loss in postmenopausal women, but bone quality data are still lacking. To clarify this, we investigated the effects of genistein compared with alendronate, raloxifene and oestradiol in an animal model of established osteoporosis.. Six months after ovariectomy, 96 ovariectomized (OVX) rats were divided into 8 equal groups, randomized to treatments (genistein aglycone (1 and 10 mg kg(-1) s.c.); alendronate (0.003 and 0.03 mg kg(-1) s.c.); raloxifene hydrochloride (0.05 and 0.5 mg kg(-1) s.c.); 17-alpha-ethinyl oestradiol (0.003 and 0.03 mg kg(-1) s.c.)) for 12 weeks. Untreated OVX (n=12) and sham OVX (n=12) were used as controls. At the beginning and end of treatment, bone mineral density (BMD) and bone mineral content (BMC) were assessed. At the end of the experiment, calcium, phosphorus, bone-alkaline phosphatase (b-ALP), collagen C-telopeptide (CTX), osteoprotegerin (OPG) and soluble receptor activator of nuclear factor-kappaB ligand (sRANKL) were assayed. Femurs were removed and tested for breaking strength and histology.. Genistein (10 mg kg(-1)) showed a greater increase in both BMD (P<0.0001 vs OVX) and BMC than all the other treatments. Moreover, genistein significantly increased breaking strength, bone quality, b-ALP (P<0.0001 vs OVX) and OPG, and reduced CTX and sRANKL compared with the other treatments at all dose levels.. The results strongly suggest that the genistein aglycone might be a new therapy for the management of postmenopausal osteoporosis in humans.

Topics: Alendronate; Animals; Bone Density; Bone Density Conservation Agents; Disease Models, Animal; Dose-Response Relationship, Drug; Estradiol; Female; Genistein; Osteoporosis; Ovariectomy; Phytoestrogens; Raloxifene Hydrochloride; Random Allocation; Rats; Rats, Sprague-Dawley; Time Factors

To investigate the preventive effect of the phytoestrogen daidzein on prostatic hyperplasia induced by testosterone in rats.. Thirty-six male adult Sprague-Dawley rats were equally randomized into 6 groups: Group I and II (normal control and model, treated with 1 ml distilled water by oral gavage), and Group III-VI (low-, medium- and high-dose daidzein and positive control, respectively given daidzein at 2, 20 and 100 mg/kg and diethylstilbestrol at 0.1 mg/kg once a day for 90 days). From the 91st day , Group III-VI were treated with subcutaneous injection of testosterone at 7.5 mg/kg/d for 10 days to induce prostatic hyperplasia. The wet weight and the index of the prostate were obtained, its morphological changes detected and the changes of ERalphaa and ERbeta expressions in the prostate observed by immunohistochemistry.. Compared with the model group, the wet weight and the index of the prostate were significantly reduced in the 3 daidzein groups and the positive control group (P < 0.05 or P < 0.01). Medium- and high-dose daidzein induced a more obvious alleviation of prostate hyperplasia, characterized by thinner epithelia, decreased secretions in the glandular cavity and reduced interstitial tissues. The expression of ERalpha showed no significant difference between the model group and the other groups, while that of ERbeta was markedly decreased in the daidzein-treated groups as compared with the normal control or the model group.. The phytoestrogen daidzein has some preventive effect on prostatic hyperplasia induced by testosterone in rats.

Topics: Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Isoflavones; Male; Phytoestrogens; Prostate; Prostatic Hyperplasia; Random Allocation; Rats; Rats, Sprague-Dawley; Testosterone

To find a more potent alternative with less oestrogen-related side effects for hormone replacement therapy (HRT) in postmenopausal osteoporosis, we designed and synthesized a NO-releasing prodrug of genistein (NO-G) according to the structure of NONSAIDs. The purpose of this study was to evaluate the effects of the prodrug on bone in ovariectomised (OVX) rats.. Forty-eight adult Sprague-Dawley female rats were ovariectomised and treated with vehicle, 9 mg/kg genistein and 4.5, 9 or 18 mg/kg NO-G by oral administration daily. The bioassays were performed in terms of bone mineral density (BMD), mechanical testing, bone formation markers, bone alkaline phosphatase (b-ALP) and osteocalcin (OCN) and bone resorption marker urine deoxypyridinoline (DPD). In addition, the effects of the drugs on uterus and body weight were examined.. After treatment for 12 weeks, the BMD of whole femur and tibia in the NO-G (9 and 18 mg/kg) groups were 12.1% and 12.2% higher than that of OVX group (P<0.01); the bending strength of the femur was 11.2% and 12.2% higher than OVX group (P<0.01). The OVX-induced increase of serum b-ALP, OCN and urinary DPD were markedly attenuated. The prodrug showed no side effects on uterus and body weight.. The NO-releasing prodrug of genistein improves the bone loss in OVX rats without stimulatory effects on the uterus, which suggests that the product could potentially be used for the prevention and treatment of postmenopausal osteoporosis.

Topics: Administration, Oral; Animals; Body Weight; Bone Density; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Genistein; Nitric Oxide; Osteoporosis; Ovariectomy; Phytoestrogens; Prodrugs; Rats; Rats, Sprague-Dawley; Treatment Outcome

Intact female rats fed a high-phytoestrogen diet are protected against adverse left ventricular (LV) remodeling induced by chronic volume overload. We hypothesized that both phytoestrogens and ovarian hormones, particularly estrogen, are necessary for this dietary-induced cardioprotection. To test this hypothesis, eight groups of female rats were studied; rats were fed either a high-phytoestrogen (+phyto) or phytoestrogen-free diet. Groups included sham-operated rats, intact rats with fistula (Fist), ovariectomized rats with fistula (Fist-OX), and Fist-OX rats treated with estrogen (EST). Myocardial function and remodeling were assessed after 8 wk of volume overload using a blood-perfused isolated heart apparatus. Fist-OX rats developed significant ventricular dilatation and increased compliance vs. intact Fist rats, which were associated with a significant decrease in contractility. Estrogen treatment prevented pulmonary edema and attenuated LV hypertrophy and dilatation but did not maintain contractility. However, dietary phytoestrogens completely prevented LV dilatation in both the Fist+phyto and Fist-OX+EST+phyto groups but had no effect on LV remodeling in the Fist-OX+phyto group. Contractility was significantly greater in the estrogen-treated rats fed the phytoestrogen diet than in those treated with estrogen alone. Dietary phytoestrogens did not affect LV or uterine mass, serum estrogen, LV estrogen receptor expression, or cardiac function in sham animals. These data indicate that estrogen is not solely responsible for the cardioprotection exhibited by intact females and that phytoestrogens can work synergistically with ovarian hormones to attenuate ventricular remodeling induced by chronic volume overload in female rats.

Topics: Animals; Aorta, Abdominal; Arteriovenous Shunt, Surgical; Cardiac Output; Cardiomegaly; Chronic Disease; Diet; Disease Models, Animal; Estrogen Replacement Therapy; Estrogens; Female; Heart Failure; Myocardial Contraction; Ovariectomy; Phytoestrogens; Rats; Rats, Sprague-Dawley; Time Factors; Venae Cavae; Ventricular Function, Left; Ventricular Pressure; Ventricular Remodeling

The aims of the present study were to assess the effects of long-term estrogen replacement therapy (ERT) on size and indices of bone turnover in periarticular osteophytes in ovariectomized cynomolgus monkeys and to compare dynamic indices of bone turnover in osteophyte bone with those of subchondral bone (SCB) and epiphyseal/metaphyseal cancellous (EMC) bone. One hundred sixty-five adult female cynomolgus macaques were bilaterally ovariectomized and randomly divided into three age- and weight-matched treatment groups for a 36-month treatment period. Group 1 (OVX control) received no treatment, Group 2 (SPE) received soy phytoestrogens, and Group 3 (ERT) received conjugated equine estrogens in the diet; all monkeys were labeled with calcein before necropsy. A midcoronal, plastic-embedded section of the right proximal tibia from 20 randomly selected animals per treatment group was examined histologically. Forty-nine of the sections (OVX control, n=16; SPE, n=16; ERT, n=17) contained lateral abaxial osteophytes, and static and dynamic histomorphometry measurements were taken from osteophyte bone, SCB from the lateral tibial plateau, and EMC bone. Data were analyzed using the ANOVA and Kruskal-Wallis test, correlation and regression methods, and the Friedman and Wilcoxon signed rank test. There was no significant effect of long-term ERT on osteophyte area or on any static or dynamic histomorphometry parameters. The bone volume, trabecular number, and trabecular thickness in osteophyte bone were considerably higher than in EMC bone; whereas, trabecular separation was considerably lower in osteophyte bone. In all three treatment groups, BS/BV was significantly lower in osteophyte bone vs. EMC bone and significantly higher in osteophyte bone vs. lateral SCB. We conclude that osteophyte area and static and dynamic histomorphometry parameters within periarticular tibial osteophytes in ovariectomized cynomolgus monkeys are not significantly influenced by long-term ERT, but that site differences in static and dynamic bone histomorphometry parameters exist, particularly between EMC and osteophyte bone.

Topics: Analysis of Variance; Animals; Bone Remodeling; Disease Models, Animal; Estrogen Antagonists; Estrogen Replacement Therapy; Estrogens; Estrogens, Conjugated (USP); Female; Humans; Macaca fascicularis; Osteoarthritis, Knee; Osteophyte; Ovariectomy; Phytoestrogens; Postmenopause; Random Allocation; Tibia

This study was aimed to examine effects of genistein on leukocyte adhesion in femur microcirculation in relation to bone-loss induced in ovariectomized female rats. Sixty-four female Wistar rats were divided into 4 groups: sham (daily treated with vehicle; DMSO, sc; 100 microl/day), ovariectomized rat treated with vehicle (OVX(veh)), 17beta-estradiol treated-ovariectomized rat (OVX(E2), 5 microg/kg/day, s.c.) and genistein treated-ovariectomized rat (0.25 mg/kg/day, s.c.; OVX(gen)). One and three weeks after the ovariectomy, blood flow perfusion (BF) in femur tissue was measured using laser Doppler flowmetry. Leukocyte adhesion in femur venules (15-30 microm in diameter) of each group was evaluated by intravital fluorescence videomicroscopy. The bone mineral content (BMC) was measured and expressed in terms of ratio of ash-to-dry matter weight. Serum osteocalcin and alkaline phosphatase levels were determined using chemiluminescence immunoassay. In both one and three week-OVX(veh), leukocyte adhesion increased significantly, compared to their age-matched sham groups, but it decreased significantly in OVX(gen), compared to OVX(veh) (p<0.05). In three week-OVX(gen), both BF and BMC increased significantly, but osteocalcin and alkaline phosphatase levels decreased, compared to those of three week-OVX(veh). In conclusion, genistein supplementation could effectively prevent bone-loss and microvascular endothelial dysfunction induced by estrogen deficiency.

Topics: Animals; Bone and Bones; Bone Density; Cell Adhesion; Disease Models, Animal; Female; Femur; Genistein; Leukocytes; Microscopy, Video; Ovariectomy; Phytoestrogens; Rats; Rats, Wistar; Venules

Inflammatory bowel disease (IBD) is very common in Europe and USA. Its incidence in East Asia has been traditionally low, albeit the risk of IBD increases in Asian immigrants adopting western lifestyles, suggesting a strong role of environmental/dietary factors in IBD. A lifelong exposure to phytoestrogen-rich diets has been associated with a decreased risk of developing breast cancer and might also be protective against IBD. We studied the influence of in utero and postnatal exposure to a phytoestrogen (PE)-rich diet on acute inflammation in an animal model of TNBS-induced colitis. Wistar rats were exposed in utero and postnatally to high (genistein: 240 microg/g feed; daidzein: 232 microg/g feed) or very low levels (genistein and daidzein <10 microg/g feed) of phytoestrogen isoflavones fed to pregnant dams with the diet and throughout nursing. After weaning, the offspring had free access to these diets. At the age of 11 weeks, colitis was induced with an enema of TNBS. After 3 days, animals were sacrificed and tissues were collected for histological evaluation and analysis of molecular markers of inflammation. Animals kept on a PE-rich diet (PRD) had higher colon weights than animals on low PE-levels (PDD), suggesting enhanced acute inflammation by phytoestrogens. This result was supported by histological findings and by analysis of myeloperoxidase activity. Interestingly, relative mRNA and protein expression of cyclooxygenase-2 (COX-2) were modulated in rats on PRD, providing evidence that COX-2, the inducible isoform of the enzyme, is involved in the management of colonic inflammation. Our results suggest that early-in-life exposure to PE might not protect against the development of IBD but enhances the extent of acute inflammation.

Topics: Acute Disease; Animal Feed; Animals; Colitis; Colon; Cyclooxygenase 2; Diet; Disease Models, Animal; Female; Organ Size; Peroxidase; Phytoestrogens; Pregnancy; Pregnancy, Animal; Prenatal Exposure Delayed Effects; Rats; Rats, Wistar; RNA, Messenger; Time Factors; Trinitrobenzenesulfonic Acid; Uterus

To evaluate the effect of estrogen replacement therapy or soy isoflavones supplement on endothelium-dependent relaxation in vitro and gene expression of endothelial nitric oxide synthase (eNOS) in cerebral arteries in a rabbit model of human hypercholesterolemia.. Thirty-six female ovariectomized Watanabe heritable hyperlipidemic (WHHL) rabbits were randomised to treatment with 17beta-estradiol (17beta-E(2)), SoyLife 150 or control for 16 weeks. Ring segments of basilar artery (BA) and posterior cerebral artery (PCA) were mounted in myographs for isometric tension recordings. Concentration response curves to carbamylcholine chloride, sodium nitroprusside (SNP) and l-NAME were evaluated after precontraction with potassium. Total RNA was extracted, reverse transcribed and eNOS quantitated by real-time polymerase chain reaction (real-time PCR).. Plasma cholesterol was significantly higher at termination in the SoyLife group (P<0.0001), whereas low-density lipoprotein (LDL) cholesterol was comparable in all treatment groups. Neither treatment influenced the endothelium-dependent responses to carbamylcholine chloride or l-NAME or the endothelium-independent response to SNP in any of the arteries. Correspondingly, eNOS mRNA was similarly expressed in all treatment groups in both arteries.. Improvement of cerebral endothelial function by estrogen or soy isoflavones in ovariectomized WHHL rabbits is not supported by the present data. The findings may be unique to the WHHL rabbit in which the hypocholesterolemic effect of estrogens mediated by upregulation of liver LDL receptors is excluded.

Topics: Animals; Basilar Artery; Cholesterol, LDL; Disease Models, Animal; Endothelium, Vascular; Estradiol; Estrogen Replacement Therapy; Female; Hyperlipidemias; Isoflavones; Nitric Oxide Synthase Type III; Ovariectomy; Phytoestrogens; Posterior Cerebral Artery; Rabbits; Random Allocation; Up-Regulation; Vasodilation

Soy products are of particular interest because of their potential health benefits in a range of hormonal conditions, such as osteoporosis, due to their high content in phytoestrogens. Because equol, the main metabolite from soy isoflavones, is thought to be powerful, the present study was designated to evaluate the bone-sparing effects of equol by either providing the molecule through the diet or by eliciting its endogenous production by modulating intestinal microflora by short-chain fructooligosaccharides (sc-FOS) or live microbial (Lactobacillus casei) together with daidzein, its precursor.. A comparison with daidzein and genistein was also performed. Rats (3 months old) were ovariectomised (OVX) or sham-operated (SH). Ovariectomised rats were randomly assigned to six experimental diets for 3 months: a control diet (OVX), the control diet supplemented with either genistein (G), or daidzein (D), or equol (E) at the level of 10 microg/g body weight/d. The remaining OVX rats were given daidzein at the dose of 10 mug/g body weight/d, simultaneously with short-chain FOS (Actilight, Beghin-Meiji) (D+FOS) or Lactobacillus casei (Actimel, Danone) (D+L). The SH rats were given the same control diet as OVX.. Genistein, daidzein or equol exhibited a bone sparing effect. Indeed, total femoral bone mineral density (BMD) was significantly enhanced (compared to that of OVX rats), as was the metaphyseal compartment. Bone strength was improved by E consumption, but not by genistein or daidzein given alone. As far as the FOS diet is concerned, the addition of prebiotics significantly raised efficiency of the daidzein protective effect on both femoral BMD and mechanical properties. The effects of lactobacillus were similar, except that the increase in metaphyseal-BMD was not significant.. In conclusion, long-term equol consumption, like genistein and daidzein, in the ovariectomized rat, provides bone sparing effects. Adding indigestible sugars, such as FOS or live microbial as L. casei, in the diet significantly improves daidzein protective effects on the skeleton.

Topics: Animals; Biomarkers; Body Weight; Bone Density; Bone Resorption; Disease Models, Animal; Equol; Female; Femur; Genistein; Glycine max; Isoflavones; Organ Size; Osteocalcin; Osteoporosis; Ovariectomy; Phytoestrogens; Rats; Rats, Wistar; Uterus

Oxidative stress is a major cause of cardiovascular tissue fibrosis. We evaluated the effects of daily doses of soy isoflavones, genistein and daidzein on cardiovascular tissue fibrosis in Otsuka Long-Evans Tokushima Fatty (OLETF) diabetic rats and Long-Evans Tokushima Otsuka (LETO) non-diabetic rats as a severe or mild oxidative stress model, respectively. Glucose and lipid metabolisms did not improve with genistein or daidzein treatment. However, genistein decreased hydroxyproline concentrations in the heart. Hydroxyproline reductions as a result of genistein were mildly stronger than those of daidzein. Thus, genistein significantly suppressed the progression of myocardial fibrosis in LETO rats despite the insignificant changes in OLETF rats. Although a daily dosage of isoflavone was not sufficient to prevent tissue fibrosis under marked oxidative stress in the early stage of diabetes, isoflavones might promise significant clinical benefits by reducing oxidative stress in the heart during aging.

Topics: Analysis of Variance; Animals; Aorta, Thoracic; Blood Glucose; Blood Pressure; Cardiovascular System; Collagen; Disease Models, Animal; Disease Progression; Fibrosis; Heart Rate; Hydroxyproline; Isoflavones; Lipid Metabolism; Male; Myocardium; Organ Size; Oxidative Stress; Phytoestrogens; Rats; Rats, Inbred OLETF; Rats, Long-Evans; Severity of Illness Index; Soy Foods

The study reported here was designed to determine whether a phytoestrogen-containing soy extract (SSE) could negate/overwhelm the inhibitory effects of ICI 182 780 on the growth of estrogen-sustained human breast cancer xenografts (MCF-7), in ovariectomized athymic mice. As expected, estradiol-supplemented tumors did not grow over the study period in ICI 182 780-treated females; concomitant administration of 50 mg/kg per day SSE slightly potentiated the inhibitory activity of the drug, while at 100 mg/kg per day, SSE partially negated ICI 182 780 activity. In keeping with these in vivo outcomes, we observed that the level of cyclin D1 (and progesterone receptor) in MCF-7 xenografts was considerably reduced by ICI 182 780, an effect enhanced by concomitant treatment with 50 SSE, but reduced by the higher dosage (i.e. 100 mg/kg per day). Thrombospondin-1 (TSP-1) and kallikrein 6 (KLK6) levels were also reduced following ICI 182 780, although to a lesser degree; again, combined anti-estrogen and SSE produced a dose-dependent regulation in TSP-1 and KLK6 tumor level, with a further reduction in the mRNA gene expression at 50 SSE (compared with ICI 182 780) and a partial reversion of the drug-induced down-regulation at 100 mg/kg per day. No modulation was detected in the serum concentration of IGF-1 (a potent mitogen for estrogen receptor-positive breast cancer cell lines) either upon treatment with ICI 182 780 or concomitant administration of the anti-estrogen with SSE. In conclusion, results from this study raise concerns about the consumption of isoflavone supplements in conjunction with ICI 182 780 therapy, in postmenopausal women with estrogen-dependent breast cancer.

Topics: Animals; Antineoplastic Agents, Hormonal; Breast Neoplasms; Cell Proliferation; Disease Models, Animal; Drug Synergism; Estradiol; Estrogens; Female; Fulvestrant; Glycine max; Humans; Kallikreins; Mice; Mice, Nude; Neoplasms, Hormone-Dependent; Organ Size; Phytoestrogens; Plant Extracts; RNA, Messenger; Thrombospondin 1; Uterus; Xenograft Model Antitumor Assays

The use of soy isoflavones is a potential alternative to hormone replacement therapy in post-menopausal bone-loss prevention. Nevertheless, phytoestrogens can target other organs and may disrupt cell proliferation, or could modify endogenous steroid hormone metabolism. These mechanisms could be linked to an increased risk of developing cancer. We therefore studied the possible side effects of such treatments in an experimental model of menopause. Forty adult female Wistar rats were ovariectomized and fed with a genistein-, daidzein- or equol-supplemented diet at bone-sparing levels (10 mg/kg BW/day) for 3 months. The estrogenic effects were assessed by histological and molecular analyses on reproductive organs. The impact on the oxidative metabolism of estradiol and on associated cytochrome P450 (CYP) activities was evaluated in liver microsomes. The relative wet weights of both the uterus and the vagina were increased in the equol group, but no significant changes in proliferating cell nuclear antigen or hormone receptor mRNA expression were noticed. In contrast, genistein and daidzein did not induce uterotrophy but caused an overexpression of estrogen receptor alpha mRNA which could correspond to a long-lasting effect of physiological concentrations of estrogens. The hepatic metabolism of estradiol was influenced by daidzein which increased the synthesis of putative mutagenic derivatives. At the same time, genistein favored estrogen 2-hydroxylation, and equol decreased 4-hydroxyestrogen production. Surprisingly, no significant alteration in hepatic CYP activities was detected. Taken together, these results demonstrate that isoflavonoid-based bone-sparing treatments are able to cause side effects on other estrogen-sensitive target organs when given in the long-term.

Topics: Animals; Bone Density; Cytochrome P-450 Enzyme System; Disease Models, Animal; Equol; Estradiol; Female; Gene Expression Regulation; Genistein; Isoflavones; Menopause; Microsomes, Liver; Organ Size; Osteoporosis; Ovariectomy; Phytoestrogens; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Uterus; Vagina

Soy phytoestrogens have been proposed as an alternative to estrogen replacement therapy and have demonstrated potential neuroprotective effects in the brain. We have shown that a high soy diet significantly reduces infarct size following permanent middle cerebral artery occlusion (MCAO). Here, we tested the hypothesis that a high soy diet would attenuate programmed cell death after stroke. Adult female Sprague-Dawley rats were ovariectomized and fed either an isoflavone-reduced diet (IFP) or a high soy diet (SP) for 2 weeks before undergoing 90 min of transient middle cerebral artery occlusion (tMCAO) followed by 22.5 h reperfusion. Infarct size, as assessed by triphenyltetrazolium chloride staining, was significantly reduced by a high soy diet (P<0.05). Apoptosis in the ischemic cortex, measured by TUNEL staining, was significantly reduced by the high soy diet. The number of active caspase-3 positive cells and caspase-mediated alpha-spectrin cleavage were also significantly decreased in the ischemic cortex of SP rats. Furthermore, nuclear translocation of apoptosis-inducing factor (AIF) was significantly reduced in the ischemic cortex of SP rats. Soy significantly increased bcl-x(L) mRNA and protein expression in the ischemic cortex compared with IFP rats. Immunohistochemistry revealed increased neuronal expression of bcl-2 and bcl-x(L) in the ischemic cortex of both IFP and SP rats following tMCAO. These results suggest that a high soy diet decreases both caspase-dependent and caspase-independent programmed cell death following tMCAO. Further, a high soy diet enhances expression of the cell survival factor bcl-x(L) following tMCAO, contributing to the neuroprotective effects of soy in the ischemic cortex.

Topics: Active Transport, Cell Nucleus; Animals; Apoptosis; Apoptosis Inducing Factor; Apoptosis Regulatory Proteins; bcl-X Protein; Brain; Caspases; Cell Survival; Disease Models, Animal; Estrogens; Female; Food, Formulated; Infarction, Middle Cerebral Artery; Nerve Degeneration; Neuroprotective Agents; Phytoestrogens; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; RNA, Messenger; Soy Foods; Stroke

Topics: Animals; Cholesterol, Dietary; Disease Models, Animal; Female; Genistein; Hypercholesterolemia; Male; Myocardial Infarction; Phytoestrogens; Rabbits; Risk Factors

Human endometrial carcinomas, as well as complex atypical hyperplasias (CAH), are estrogen related and frequently have mutations in the PTEN gene. However, the mutual contribution of estrogen and PTEN mutations to endometrial carcinogenesis in vivo is unknown. To address this issue, we investigated whether neonatal estrogenic treatments augment the incidence of CAH and carcinomas in murine PTEN (mPTEN) heterozygous (+/-) mutant mice, an animal model for endometrial carcinoma. Low doses of diethylstilbestrol (1 ng/g/day), genistein (50 microg/g/day) in phytoestrogens, estriol (E(3)) (4 microg/g/day), and vehicle (ethanol and corn oil) were administered subcutaneously daily to neonatal pups from the 1st to 5th day after birth. At 52 weeks of age, the morphological changes in the endometrium, and uterine expression of Hoxa 10 and Hoxa 11, were evaluated. These Hoxa genes are abdominal B-type homeobox genes, which normally regulate differentiation of the Müllerian duct. The incidence of CAH and adenocarcinomas of the endometrium was significantly decreased by the neonatal estrogenic treatments in the mPTEN+/- mice. Coincidentally, all treatments significantly decreased the stromal cell density, and CAH and adenocarcinomas rarely developed in the epithelium adjacent to the affected endometrial stroma. Moreover, the uterine expression of Hoxa 10 in mice with neonatal genistein and E(3) treatments, and that of Hoxa 11 in mice with all treatments, was significantly lower when compared with vehicle alone. Taken together, neonatal estrogenic exposure induced stromal atrophy and/or hyalinization accompanied by repressed expression of Hoxa 10 and Hoxa 11, and exerted an inhibitory effect on PTEN-related tumorigenesis. These findings provide new insight into the interaction between endometrial epithelium and stroma in endometrial carcinogenesis in vivo.

Topics: Adenocarcinoma; Animals; Animals, Newborn; Disease Models, Animal; Endometrial Neoplasms; Female; Gene Expression Regulation, Neoplastic; Homeobox A10 Proteins; Homeodomain Proteins; Mice; Mice, Inbred C57BL; Organ Size; Phytoestrogens; PTEN Phosphohydrolase; Recombination, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Uterus

As phytoestrogens are postulated as being neuroprotectants, we assessed the hypothesis that dietary isoflavone-type phytoestrogens are neuroprotective against ischemic stroke. Transient focal cerebral ischemia (90 min) was induced by middle cerebral artery occlusion (MCAO) following the intraluminal thread technique, both in rats fed with soy-based diet and in rats fed with isoflavone-free diet. Cerebro-cortical laser-Doppler flow (cortical perfusion, CP), arterial blood pressure, core temperature, PaO2, PaCO2, pH and glycemia were measured before, during and after MCAO. Neurological examination and infarct volume measurements were carried out 3 days after the ischemic insult. Dietary isoflavones (both glycosides and aglycones) were measured by high-performance liquid chromatography. Neither pre-ischemic, intra-ischemic nor post-ischemic CP values were significantly different between the soy-based diet and the isoflavone-free diet groups. Animals fed with the soy-based diet showed an infarct volume of 122 +/- 20.2 mm3 (19 +/- 3.3% of the whole ipsilateral hemisphere volume). In animals fed with the isoflavone-free diet the mean infarct volume was significantly higher, 191 +/- 26.7 mm3 (28 +/- 4.1%, P < 0.05). Neurological examination revealed significantly higher impairment in the isoflavone-free diet group compared with the soy-based diet group (3.3 +/- 0.5 vs. 1.9 +/- 0.5, P < 0.05). These results demonstrate that dietary isoflavones improve stroke outcome after transient focal cerebral ischemia in such a way that a higher dietary isoflavone content results in a lower infarct volume and a better neurological status.

Topics: Animals; Body Weight; Cerebral Infarction; Chromatography, High Pressure Liquid; Diet; Disease Models, Animal; Ischemic Attack, Transient; Laser-Doppler Flowmetry; Male; Neurologic Examination; Neuroprotective Agents; Phytoestrogens; Rats; Rats, Wistar; Reperfusion; Stroke; Time Factors

Reduced estrogen levels occurring during menopause in woman are accompanied by a variety of disorders, e. g., hot flushes, depressions, osteoporosis, increase of body weight, and reduced movement drive. In this study we investigated the combined effects of physical activity, estradiol substitution, and a phytoestrogen-rich diet on bone mineral density, increase of body weight, and movement drive in an animal model. Ovariectomized (OVX) female Wistar rats were either fed an isoflavone-rich diet (IRD) or substituted with 17beta-estradiol (E2) for 3 months. Sham-operated rats (Sham) and vehicle-treated OVX animals served as controls. One half of the animals had the opportunity of voluntary wheel running. OVX rats displayed an eight times lower movement activity than Sham animals. E2 treatment, but not IRD, significantly increased the movement activity of OVX rats. During 3 months the lowest increase of body weight was observed in Sham animals, the highest rate in OVX animals. Along with running activity E2 treatment, but not IRD, also lowered the increase of body weight significantly compared to OVX animals. Bone mineral density (BMD) in the trabecular area of the tibia was strongly reduced in OVX rats compared to Sham animals. In contrast to IRD, E2 substitution resulted in a protection of BMD in this area compared to OVX animals. Our data demonstrate that body weight, movement drive, and BMD are positively influenced by E2. The steroid estrogen acts in the trabecular area of the tibia in a bone-protective manner, increases movement drive and antagonizes the increase of body weight. All these effects could not be observed in animals fed an isoflavone-rich diet.

Topics: Animals; Body Weight; Bone Density; Dietary Supplements; Disease Models, Animal; Estradiol; Female; Isoflavones; Motor Activity; Osteoporosis; Ovariectomy; Phytoestrogens; Phytotherapy; Rats; Rats, Wistar

There is a great deal of controversy surrounding the issue of hormone replacement therapy after transplantation. The question whether or not this therapy has effects in cardiac allograft vasculopathy (CAV), the Achilles heel of cardiac transplantation or other unique aspects of allograft function is still unknown.. We investigated the long-term effect of 17beta-estradiol as well as phytoestrogen Coumestrol, a synthetically produced phytoestrogen, on the development of CAV and the degree of fibrosis in an ovariectomized female heterotopic chronic allograft model (LEW-F344).. We found that, 150 days after transplantation, no significant effect of estrogen application on intimal thickening of coronary arteries was observed. 17beta-estradiol and phytoestrogen Coumestrol did significantly reduce the perivascular immune reaction. However, the immune effect had no consequence on the intensity of CAV. Although neither 17beta-estradiol nor phytoestrogen Coumestrol revealed a positive effect on CAV, the group of animals treated with 17beta-estradiol showed the highest decline in heart function and the most distinct fibrosis.. 17beta-estradiol does not affect CAV positively, but worsens cardiac allograft function and leads to increased fibrosis. This is the first study showing a negative effect of 17-beta-estradiol after heart transplantation in the long term.

Topics: Animals; Body Weight; Coumestrol; Disease Models, Animal; Estradiol; Female; Heart Transplantation; Phytoestrogens; Postoperative Complications; Rats; Rats, Inbred F344; Rats, Inbred Lew; T-Lymphocytes; Transplantation, Homologous; Transplantation, Isogeneic; Uterus; Weight Gain

Long-term maintenance of ovariectomized rats (9 weeks) on chow containing high phytoestrogen levels (Purina LabDiet 5001) as compared to chow with minimal phytoestrogens (Harlan 2016 Teklad) was associated with better performance of the spatial memory task, object placement, increased dendritic spine density in CA1 and prefrontal cortex pyramidal neurons, and higher uterine weights. Object recognition memory, anxiety on an elevated plus maze and body weight were unaffected by phytoestrogen levels in the diet.

Topics: Administration, Oral; Animals; Anxiety; Dendritic Spines; Disease Models, Animal; Estrogens; Female; Food, Formulated; Hippocampus; Maze Learning; Memory; Memory Disorders; Neural Pathways; Neuronal Plasticity; Organ Size; Phytoestrogens; Prefrontal Cortex; Rats; Synapses; Treatment Outcome; Up-Regulation; Uterus

There is increasing interest in the role of complementary and alternative medicines for the treatment of menopause-related problems. This study compared the preventive effect on atheroma formation of a commercially available mixed phytoestrogen concentrate with that of estradiol.. An ovariectomized cholesterol-fed rabbit model of atheroma formation was used. Rabbits were ovariectomized before the commencement of the 12-week treatment period. There were two control groups. Control Group 1 received isoflavone-free rabbit chow whilst Control Group 2 received 1% cholesterol-enriched isoflavone-free rabbit chow. Rabbits in Group 3 received 1% cholesterol-enriched isoflavone-free rabbit chow plus a 500 mg tablet containing a concentrated extract of Trifolium pretense (red clover). Rabbits in Group 4 received 1% cholesterol-enriched isoflavone-free rabbit chow plus a 0.5 mg tablet of oral estradiol. Atheroma formation was measured by, first, calculation of the area of atheroma on the intimal surface, and, second, measuring the cholesterol content in the aorta.. There were no significant differences in serum cholesterol between the cholesterol-fed control Group 2 and the treatment Groups 3 and 4. However, there was significantly less staining for atheroma and significantly less cholesterol accumulation in the aorta in Group 4 (estradiol-treated) rabbits compared with either control Group 2 or Group 3 (phytoestrogen-treated) rabbits.. In this study, only estradiol was shown to have a significant protective effect against atheroma formation.

Topics: Administration, Oral; Animals; Aorta; Atherosclerosis; Cholesterol; Cholesterol, Dietary; Disease Models, Animal; Estradiol; Female; Ovariectomy; Phytoestrogens; Rabbits; Random Allocation

The mammalian lignan enterolactone (ENL) produced from plant lignans, e. g. secoisolariciresinol diglycoside (SDG), may protect against various cancers in humans. The present work aims to evaluate the effect of flaxseed on tumour formation in multiple intestinal neoplasia (Min) mice, a model for colon tumorigenesis.. Male and female Min mice were fed either with a non-fibre control diet or the same diet supplemented with 0.5 % (w/w) defatted flaxseed meal. Conversion of SDG to the mammalian lignans enterodiol (END) and ENL in the gut, and plasma ENL, were measured by HPLC with coulometric electrode array detector (CEAD) and timeresolved fluoroimmunoassay, respectively. Wild-type mice were also fed with the experimental diets in order to see whether lignan metabolism is different in Min and wild-type mice.. The total number of adenomas or their size in the small intestine was not different in the flaxseed and control groups. The flaxseed group had a tendency for a decreased number of colon adenomas in both genders. Gender and genotype based differences were found in the intestinal ENL levels. When compared to Min females, Min males in the flaxseed group had several fold higher ENL levels in the small intestine (Min males 125 +/- 124.5 nmol/g vs. females 22.8 +/- 16.0 nmol/g, P = 0.048) and caecum (47.6 +/- 31.6 nmol/g vs. females 14.5 +/- 6.6 nmol/g, P = 0.001). Presence of adenomas in the gut influences the intestinal lignan metabolism. Min mice had less intestinal END and ENL as compared with the wild-type mice (P < 0.05). Mean plasma ENL increased 7-fold during the flaxseed feeding (7 nmol/L in control vs. 50 nmol/L in flaxseed group) but no differences between gender and genotype were found. The plasma ENL level did not correlate with adenoma number in the small intestine and colon.. The number of intestinal adenomas in the Min mouse model is not related to ENL level in plasma nor is it associated with the levels of intestinal lignans. A gender difference in ENL lignan metabolism was found in the gut but not in the plasma.

Topics: Adenoma; Animals; Chromatography, High Pressure Liquid; Disease Models, Animal; Female; Flax; Fluoroimmunoassay; Genotype; Intestinal Neoplasms; Lignans; Male; Mice; Mice, Mutant Strains; Neoplasms, Multiple Primary; Phytoestrogens; Random Allocation; Sex Factors

Epidemiological studies suggest that post-menopausal hormone replacement therapy (HRT) reduces colorectal cancer (CRC) incidence. Phytoestrogens, including the soy isoflavone genistein and coumestrol, are used by many women as alternatives to HRT. Previous studies showed that ovariectomy induced a 77% increase in intestinal adenoma number in the C57BL/6J-Min/+ (Min/+) mouse, an animal model of adenomatous polyposis coli (APC)-associated CRC. Replacement of estradiol (E(2)) in ovariectomized Min/+ mice reduced tumor number to baseline and up-regulated the expression of estrogen receptor beta (ERbeta). We hypothesized that the phytoestrogens genistein and coumestrol would inhibit intestinal tumorigenesis in ovariectomized Min/+ mice. Min/+ and Apc(+/+) (WT) mice were ovariectomized and assigned to either a control diet or treatment with E(2), genistein or coumestrol. Treatment of ovariectomized Min/+ (Min/+ OX) mice with genistein resulted in a non-significant reduction in tumor number. Min/+ OX mice treated with coumestrol had significantly fewer tumors than untreated Min/+ OX controls and the same number of tumors as non-ovariectomized Min/+ mice. Bromodeoxyuridine migration assays also demonstrated that treatment with E(2) or coumestrol improved enterocyte migration rate. Immunoprecipitation and immunohistochemistry analyses showed that impaired association of the adherens junction proteins E-cadherin and beta-catenin in Min/+ mice was improved by treatment with either E(2) or coumestrol. Immunoblot analyses also showed that expression of ERbeta was elevated in enterocytes of Min/+ OX mice treated with E(2) or coumestrol as compared with those of untreated Min/+ OX mice. In conclusion, both coumestrol and E(2) prevent intestinal tumorigenesis and ameliorate enterocyte migration and intercellular adhesion in the Apc(Min/+) mouse model of CRC.

Topics: Animals; Cell Movement; Colorectal Neoplasms; Coumestrol; Disease Models, Animal; Female; Genes, APC; Genistein; Immunohistochemistry; Immunoprecipitation; Intestinal Mucosa; Intestines; Mice; Mice, Inbred C57BL; Ovariectomy; Phytoestrogens; Receptors, Estrogen

Clinical intervention studies and experimental studies with lignan-rich diets suggest that lignans may have inhibitory effects on prostate cancer, but no clinical or experimental studies with purified lignans have been published. The purpose of this study was to investigate the effect of a plant lignan 7-hydroxymatairesinol (HMR) on LNCaP human prostate cancer xenografts in athymic mice. Athymic nude male mice were injected subcutaneously with LNCaP cells. Starting 3 days after tumor cell injections, a control diet or a control diet supplemented with 0.15% or 0.30% of HMR was administered to mice and the tumor take rate and growth was observed for 9 weeks. HMR diet inhibited the growth of LNCaP tumors. Mice treated with HMR had smaller tumor volume, lower tumor take rate, increased proportion of nongrowing tumors, and higher tumor cell apoptotic index compared with controls. Furthermore, the cell proliferation index was reduced in mice receiving the 0.30% HMR diet compared with mice receiving the control diet. Our results suggest that dietary HMR started at the early phase of the tumor development inhibits the growth of the LNCaP human prostate cancer xenografts in athymic male mice.

Topics: Animals; Anticarcinogenic Agents; Apoptosis; Carcinoma; Cell Proliferation; Diet; Disease Models, Animal; Humans; Isoflavones; Lignans; Male; Mice; Mice, Nude; Models, Biological; Neoplasm Transplantation; Phytoestrogens; Prostate-Specific Antigen; Prostatic Neoplasms; Time Factors; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Dietary phytoestrogens are promoted as alternatives to synthetic estrogens for hormone therapy, however, their effects on the reproductive axis have not been exhaustively studied in vivo. Female aromatase knockout mouse (ArKO) mice are estrogen-free, anovulatory, and have a block in folliculogenesis, hemorrhagic cysts, and development of Sertoli cells within their ovaries. The purpose of this study was to evaluate the (ArKO) mouse as a model to test the effects of phytoestrogen replacement in vivo.. We examined the effects of phytoestrogen-supplemented diets on the reproductive organ weights, ovarian morphology, gonadotropin levels, and the transcript levels of ovarian somatic cell and steroidogenic markers of wild-type and ArKO mice.. The genistein diet significantly increased uterine and ovarian weights of ArKO mice. The soy, and to a larger extent the genistein diet, improved ovarian morphology. Morphological Sertoli cell transformation in ArKO mice was decreased by both diets, whereas the gene expression of Sertoli cell markers was not affected. The soy diet increased both gonadotropins in both genotypes compared with those animals on the soy-free diet. The genistein diet reduced FSH levels in ArKO mice, correlating with increased ovarian inhibin subunit expression.. Phytoestrogens are estrogenic in ArKO mice. Specifically, they can affect serum gonadotropin levels, and offset the development of Sertoli cells and hemorrhagic cysts within the ovaries. However, the effects on the mouse ovary depended on the type of dietary phytoestrogen. Further studies using the ArKO mouse are required to determine the effective doses and treatment regimes for phytoestrogens as endocrine modulators.

Topics: Animals; Diet; Disease Models, Animal; DNA Primers; Estrogen Replacement Therapy; Estrogens; Female; Gene Expression; Genistein; Gonadotropins; Inhibins; Isoflavones; Menopause; Mice; Mice, Inbred AKR; Mice, Knockout; Ovary; Phenotype; Phytoestrogens; Phytotherapy; Polymerase Chain Reaction; Soybean Proteins; Specific Pathogen-Free Organisms; Uterus

Coumestrol is a naturally occurring plant estrogen. As estrogen influences cellular and humoral immunity, and has known effects on murine models of lupus, we investigated the effect of coumestrol on disease expression in the NZB/W F1 mouse. Female NZB/W F1 mice were fed a "standard" rodent diet including soy proteins, a non-soy diet, or a non-soy diet with 0.01% coumestrol. Outcome measures included survival, autoantibody expression, immunoglobulin levels, proteinuria, renal histology and B cell immunohistochemistry, and renal mRNA expression. At 24 weeks, the treatment group had decreased prevalence of autoantibodies detected by immunofluorescence and less splenomegaly. At 39 weeks, the prevalence of autoantibodies was similar but the treatment group had less proteinuria. Overall, there was little effect of treatment on renal mRNA levels as assessed by gene array analysis, but functional ontology mapping revealed that genes encoding proteins involved in the immune response were most often affected. These results suggest that treatment with coumestrol may ameliorate some aspects of disease progression in this model of systemic autoimmunity.

Topics: Animals; Antibody Formation; Autoantibodies; Autoimmunity; Chromatin; Coumestrol; Disease Models, Animal; Female; Gene Expression; Immunoglobulins; Kidney; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Oligonucleotide Array Sequence Analysis; Phytoestrogens; RNA, Messenger

Topics: Animals; Disease Models, Animal; Estrogens, Non-Steroidal; Hot Flashes; Humans; Isoflavones; Menopause; Phytoestrogens; Phytotherapy; Plant Preparations; Sheep

Exogenous and dietary estrogens have been associated with modification of breast cancer risk. Mammary cancer model systems can be used to explore interactions between specific transgenes, and hormonal and dietary factors. Transgenic mice bearing the rat wild-type erbB-2 gene were used to study the effects of short-term hormonal exposure [17beta-estradiol (E2) or tamoxifen] or a soy meal diet on mammary carcinogenesis. In mice fed a casein diet, mammary tumors developed at an earlier age after short-term E2 exposure during the early reproductive period. The median mammary tumor latency was shortest (29 weeks) for the high-dose estrogen as compared with the lowest dose of E2 treated or placebo control mice (33 and 37 weeks, respectively). The timing of short-term E2 exposure was also important, with the most significant changes observed in mice exposed to E2 between 8 and 18 weeks of age. E2 exposure was associated with the subsequent development of more aggressive tumors as determined by histologic grade, multifocal tumor development, stromal invasion, and pulmonary metastasis. In contrast, short-term tamoxifen-exposed mice generally failed to develop mammary tumors by 60 weeks of age. Mice fed a soy meal diet developed mammary tumors at a later age than casein-fed animals treated with E2 or placebo, whereas no differences were observed by diet for the tamoxifen-treated mice. Mammary tumor prevention was >80% in tamoxifen-treated mice on either diet. Novel histologic tumor types were identified, suggesting greater phenotypic diversity than described previously. Benign mammary gland morphogenesis was also significantly altered by short-term hormonal exposure or dietary factors, consistent with the modification of mammary tumor risk in specific treatment groups. Estrogenic modulation of the mammary tumor phenotype in wild-type erbB-2 transgenic mice was observed. Histologic tumor types and clinical aggressivity not reported previously in this transgenic model were noted, suggesting greater biologic heterogeneity than reported previously. In addition, dietary phytoestrogens modified mammary development and tumor latency, suggesting a need for greater stringency in dietary assignment for transgenic mouse models of mammary neoplasia.

Topics: Age Factors; Animals; Anticarcinogenic Agents; Cocarcinogenesis; Diet; Disease Models, Animal; Estradiol; Estrogen Antagonists; Estrogens, Non-Steroidal; Female; Genes, erbB-2; Isoflavones; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Mammary Tumor Virus, Mouse; Mice; Mice, Transgenic; Phytoestrogens; Plant Preparations; Risk Factors; Tamoxifen

To determine the effects of long-term estrogen replacement therapy (ERT) on the severity of osteoarthritis (OA) of the knee joint in surgically postmenopausal (bilaterally ovariectomized) female monkeys. A secondary aim was to evaluate the effect of soy phytoestrogen (SPE) treatment on the severity of OA.. Feral adult female cynomolgus macaques were ovariectomized bilaterally and then randomly divided into 3 age- and weight-matched treatment groups. For 3 years, the first group received ERT with conjugated equine estrogens, the second group received SPE, and the third group received no treatment (controls). At necropsy, histologic lesions of OA were graded, and the area and thickness of cartilage and subchondral bone were measured. The data were summarized by principal components analysis, and the resulting factors and individual variables were compared using analysis of variance and analysis of covariance (age and weight as covariates).. Cartilage lesions of OA were significantly less severe in the animals given ERT compared with those in the control group. This treatment effect remained significant when adjusted for age and weight. The factor representing subchondral bone was significantly higher, but the number of osteophytes was lower, in the ERT group compared with the control group. SPE treatment had no significant effect on cartilage or bone lesions of OA.. These results demonstrate that long-term ERT significantly reduces the severity of OA lesions in this animal model.

Topics: Animals; Bone and Bones; Cartilage, Articular; Disease Models, Animal; Estrogen Replacement Therapy; Estrogens, Non-Steroidal; Female; Isoflavones; Macaca fascicularis; Osteoarthritis, Knee; Ovariectomy; Phytoestrogens; Plant Preparations; Random Allocation

The mortality of clinical prostate cancer is lower in Asian populations than in American or European men. Asian men typically consume more soy than their Western counterparts, leading to the investigation of individual components, particularly phytoestrogens, as protective factors against prostate cancer. Genistein, the predominant isoflavone in soy, has been reported to reduce the incidence of prostate cancer in animal models, but the underlying biological action remains to be elucidated. The purpose of this investigation was to identify the effects of the phytoestrogen, genistein and the synthetic estrogen diethylstilbestrol (DES), as a control, on development and function of the rat dorsolateral prostate (DLP) when given in the diet. The effects of testosterone and dihydrotestosterone (DHT) injections were also tested. Analysis of individual lobes of the DLP revealed that 1000 mg/kg, but not 250 mg/kg, of a genistein AIN-76A diet slightly reduced lateral prostate type 1 (LP1) bud perimeter. However, expression of the secretory dorsal protein 1 (DP1) and 5alpha-reductase type II activity were not altered in the prostate. This suggested that prostate differentiation, and not toxicity, had occurred. DES in the diet reduced and testosterone injections elevated relative prostate weights and perimeters of the dorsal, LP1, lateral prostate type 2 and DP1 expression. DHT increased relative prostate weights but did not significantly increase individual lobe perimeter. Unlike DES, maximally tolerated doses of genistein in the diet were not toxic to the rat prostate.

Topics: 3-Oxo-5-alpha-Steroid 4-Dehydrogenase; Androgen-Binding Protein; Animals; Anticarcinogenic Agents; Diethylstilbestrol; Dihydrotestosterone; Disease Models, Animal; Dose-Response Relationship, Drug; Estrogens, Non-Steroidal; Genistein; Glycine max; Growth Inhibitors; Humans; Isoflavones; Male; Organ Size; Phytoestrogens; Plant Preparations; Prostate; Prostatic Neoplasms; Rats; Rats, Sprague-Dawley; Testosterone

To evaluate the effect of estrogen replacement therapy (ERT) on the functional characteristics of coronary and cerebral arteries in a new rabbit model for postmenopausal vascular function.. Female ovariectomized Watanabe heritable hyperlipidemic (WHHL) rabbits were randomized to treatment for 16 weeks with either 17 beta-estradiol or placebo. The chow used was semi-synthetic, thereby avoiding the influence of phytoestrogens. Ring segments of cerebral and coronary arteries were mounted for isometric tension recordings in myographs. The passive and active length-tension relationships for electromechanical (high potassium), pharmacomechanical (histamine) and combined electro- and pharmacomechanical (high potassium plus histamine) contraction were evaluated.. Treatment with 17 beta-estradiol significantly changed the active length-tension relationship for the electromechanical response in the proximal coronary arteries. No changes were observed for the passive length-tension relationships.. Long-term treatment with 17 beta-estradiol lowered the electromechanical tonus of atherosclerotic coronary arteries proximally, where the atherosclerosis is most developed. This could be one of the mechanisms behind the putative protective effect of hormone replacement therapy against ischemic heart disease. The study presents a promising new animal model for the investigation of postmenopausal coronary and cerebral artery function. The data correspond well with epidemiological observations in postmenopausal women.

Topics: Animal Nutritional Physiological Phenomena; Animals; Brain; Cerebral Arteries; Coronary Artery Disease; Coronary Circulation; Coronary Vessels; Disease Models, Animal; Estradiol; Estrogens, Non-Steroidal; Female; Hyperlipidemias; Isoflavones; Myography; Phytoestrogens; Plant Preparations; Postmenopause; Rabbits; Random Allocation

The effects of 17beta-estradiol (17beta-E(2)) or the phytoestrogen naringenin on spontaneous atherosclerosis were studied in 36 ovariectomized homozygous Watanabe heritable hyperlipidemic (WHHL) rabbits receiving a semisynthetic control diet; this diet added 0.0040% 17beta-E(2;) or 0.20% naringenin, for 16 weeks. The uterine weight was increased (P < 0.001) and the concentration of estrogen receptor alpha was decreased (P < 0.001) in the 17beta-E(2) group compared with the controls. Total plasma cholesterol and triglycerides were not different from those in the controls. In lipoproteins, HDL cholesterol was increased (P < 0.01), and LDL triglyceride and IDL triglyceride were lowered (P < 0.05). The oxidation (as concentration of malondialdehyde) was increased in LDL (P < 0.05) but not in plasma. The cholesterol accumulation was decreased (P < 0.05) in the ascending aorta and in the total aorta but the ratio of intima to media and area of intima in ascending, thoracic, and abdominal aorta were not significantly different. In the naringenin group the only differences, compared with the control group, were increased HDL cholesterol (P < 0.001) and decreased activity of glutathione reductase (P < 0.05). In conclusion, 17beta-E(2), but not naringenin, attenuated aortic cholesterol accumulation independently of plasma and LDL cholesterol. Further, these results support previously suggested pro-oxidant ability of 17beta-E(2) toward LDL and a possible connection between the pro-oxidant nature of 17beta-E(2) and its antiatherogenic effect.

Topics: Animals; Aorta; Arteriosclerosis; Cholesterol; Disease Models, Animal; Erythrocytes; Estradiol; Estrogens, Non-Steroidal; Female; Flavanones; Flavonoids; Food, Formulated; Humans; Isoflavones; Lipoproteins; Molecular Structure; Ovariectomy; Oxidation-Reduction; Phytoestrogens; Plant Preparations; Rabbits

The effect of phytoestrogen intake in combination with estrogen replacement therapy (ERT) on atherogenesis is largely unknown. The aim was thus to study the impact of phytoestrogens alone, or combined with oral estrogen, on experimental atherosclerosis in cholesterol-fed rabbits.. Two separate studies were performed in ovariectomized, cholesterol-fed female rabbits. In Study A, 45 rabbits were randomized to either a soy-free diet with or without oral 17 beta-estradiol (E2) 4 mg/day, or a soy-rich diet without any hormone for 14 weeks. In Study B, 100 rabbits were randomized into five groups (oral E2 0.5, 1, 2 or 4 mg/day, or no hormone) based on a soy-rich diet for 30 weeks.. By the end of treatment in Study A, aortic cholesterol content was twice the amount in the group treated with the soy-free diet compared with the soy-rich group and with the soy-free plus E2 group (p < 0.001). In Study B, aortic cholesterol content showed no significant difference between the groups (ANOVA, p = 0.49), but a tendency towards a lower aortic cholesterol content in the E2-treated animals compared with placebo was observed.. Dietary phytoestrogens significantly reduce aortic cholesterol content with a potency comparable to that of ERT, and seem to enhance (although mildly) the antiatherogenic effect of E2 in this model.

Topics: Animals; Aorta; Cholesterol; Chromatography, High Pressure Liquid; Coronary Artery Disease; Diet; Disease Models, Animal; Estradiol; Estrogens, Non-Steroidal; Female; Genistein; Glycine max; Isoflavones; Lipoproteins; Ovariectomy; Phytoestrogens; Plant Preparations; Rabbits; Random Allocation; Uterus

Phytoestrogens are a normal constituent of soy protein and have been shown to have anti-inflammatory activity in various in vitro and in vivo models. The present study was designed to determine if a diet enriched in the phytoestrogen isoflavones, genistin and daidzin, would alter the antigen-induced cellular infiltration, particularly eosinophilia, characteristic of a guinea pig model of asthma. Throughout the duration of the study, guinea pigs were maintained on a control diet (standard guinea pig chow) or the same diet enriched in isoflavones. The animals were placed on the diet 2 weeks prior to active sensitization with ovalbumin (OA). Three weeks after sensitization, animals were challenged with OA aerosol. The cellular infiltration into the lung and protein and red blood cells (RBC) in the bronchoalveolar lavage fluid (BAL) were determined 17 hr later. In animals maintained on the control diet, OA aerosol challenge resulted in the expected increase in eosinophils in both the BAL and the lung tissue, an increase in neutrophils in the BAL, and an increase in protein and the number of RBC in the BAL. In contrast, in animals maintained on the isoflavone diet, the OA-induced eosinophilia in the lung tissue was significantly attenuated. In addition, OA challenge caused a greater increase in BAL protein in animals maintained on the isoflavone diet compared with animals on the control diet. Our results indicated that a diet enriched in isoflavones results in reduced antigen-induced eosinophilia in the lung in the guinea pig model of asthma. However, this beneficial anti-inflammatory effect of dietary phytoestrogens is accompanied by a potentially detrimental increase in antigen-induced leakage of protein into the airspace.

Topics: Aerosols; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Diet; Disease Models, Animal; Eosinophils; Erythrocytes; Estrogens, Non-Steroidal; Female; Guinea Pigs; Immunoglobulin G; Isoflavones; Lung; Ovalbumin; Phytoestrogens; Plant Preparations; Proteins

Because of their beneficial effects on atherosclerosis and cancer risk, isoflavones may be useful as a dietary alternative or supplement to postmenopausal hormone replacement therapy. Isoflavones belong to the most important phytoestrogens. They may have estrogenic and antiestrogenic effects. We examined this in a well-characterised primate model of the postmenopause.. Adult, surgically postmenopausal female macaques were treated with isoflavones, estradiol or placebo for 6 months. After 6 months of therapy histopathological, morphometrical, and immunohistochemical measurements of endometrium and mammary glands were performed.. 6 months of isoflavone-therapy did not induce proliferation or any other clinical important changes in endometrium and mammary tissue. Phytoestrogentherapy did not show estradiol comparable effects. We are discussing our results with the results of other studies, which are sometimes in contrast.. Our results indicate that isoflavones do not have estrogenic effects in the tissues studied.

Topics: Animals; Disease Models, Animal; Endometrium; Estradiol; Estrogen Replacement Therapy; Estrogens, Non-Steroidal; Female; Humans; Isoflavones; Macaca fascicularis; Mammary Glands, Animal; Phytoestrogens; Plant Preparations; Postmenopause; Soybean Proteins; Uterus