trans-sodium-crocetinate and Disease-Models--Animal

Saffron (Crocus savitus) is a Middle-Eastern herb with strong antioxidant properties. Its major constituents, safranal, crocin, and crocetin, are also antioxidants and bear structural similarities to other well-known natural antixodant substances, such as zeaxanthin. Given the role of oxidative stress in many diseases, considerable interest has been shown into the potential role of saffron supplementation as a treatment for a range of diseases. In vitro and animal studies have provided evidence that saffron and its constituents may be potent therapies for a range of pathologies, including Alzheimer's disease, age-related macular degeneration (AMD) and cardiac ischemia. Whether these findings translate into clinical efficacy, however, has as of yet been incompletely assessed. This makes assessing the role of saffron supplementation in these diseases difficult. Here, we review the current human clinical evidence supporting saffron supplementation as a treatment for a range of pathologies and the underlying science supporting its use.

Topics: Affect; Animals; Antioxidants; Cardiovascular System; Carotenoids; Clinical Trials as Topic; Cognition; Crocus; Cyclohexenes; Disease Models, Animal; Humans; Oxidative Stress; Phytotherapy; Plant Preparations; Reproduction; Terpenes; Vision, Ocular; Vitamin A; Zeaxanthins

Topics: Animals; Blood Pressure; Carotenoids; Disease Models, Animal; Heart Rate; Humans; Male; Oxygen Consumption; Oxygen Inhalation Therapy; Rats; Rats, Sprague-Dawley; Shock, Hemorrhagic; Vitamin A

Cardiovascular disease (CVD) is a chronic disease and causes the highest rate of death globally. CVD-related deaths account for 80% of all deaths in low and middle-income countries, such as China. Crocetin (CT), a carotenoid phytoconstituent already confirm their anti-inflammatory and antioxidant effects in various diseases animal models. In the study, we make effort to access the cardio-protective effect of Crocetin against vitamin D3 and high fat induced atherosclerosis in rats and scrutinize the underlying mechanism. Sprague Dawley (SD) rats were used in this study and rats were divided into different groups and high fat diet and vitamin D was used for induction the atherosclerosis. The rats were received oral administration of crocetin (5, 10 and 15 mg/kg) and simvastatin (0.5 mg/kg) until 30 days. At the end of the experimental period, lipid, cardiac markers, anti-inflammatory, antioxidant, pro-inflammatory cytokines and atherogenic index were estimated. The mRNA expression of Intercellular adhesion molecule-1 (ICAM-1), Monocyte Chemoattractant Protein-1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) in aortic tissue of the atherosclerotic rats. Crocetin significantly reduced the aortic membrane thickness and platelet aggregation rates. Crocetin also dose-dependently reduced total cholesterol (TC), very low-density lipoprotein (VLDL), triacylglycerol (TG), low-density lipoprotein (LDL) and augmented the level of high-density lipoprotein (HDL) level. Additionally, Crocetin significantly (p < 0.001) abridged the level of malonaldehyde (MDA) and augmented the level of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and glutathione peroxidase (GPx). Furthermore, Crocetin significantly (p < 0.001) dose-dependently reduced the levels of pro-inflammatory cytokines and inflammatory mediators. Crocetin attenuated mRNA expression of VCAM-1, ICAM-1 and MCP-1. Crocetin had anti-atherosclerosis and cardio-protective effects on vitamin D3 and high fat induced atherosclerosis in rats through anti-inflammatory and antioxidant mechanisms.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Antioxidants; Aorta; Atherosclerosis; Carotenoids; Chemokine CCL2; Cholecalciferol; Cytokines; Diet, High-Fat; Disease Models, Animal; Inflammation Mediators; Intercellular Adhesion Molecule-1; Lipid Metabolism; Phytotherapy; Rats, Sprague-Dawley; Vascular Cell Adhesion Molecule-1; Vitamin A

Parkinson's disease (PD) is a common neurodegenerative disorder with motor symptoms linked to the loss of dopaminergic neurons in the brain. α-Synuclein is an aggregation-prone neural protein that plays a role in the pathogenesis of PD. In our previous paper, we found that saffron; the stigma of Crocus sativus Linné (Iridaceae), and its constituents (crocin and crocetin) suppressed aggregation of α-synuclein and promoted the dissociation of α-synuclein fibrils in vitro. In this study, we investigated the effect of dietary saffron and its constituent, crocetin, in vivo on a fly PD model overexpressing several mutant α-synuclein in a tissue-specific manner. Saffron and crocetin significantly suppressed the decrease of climbing ability in the Drosophila overexpressing A30P (A30P fly PD model) or G51D (G51D fly PD model) mutated α-synuclein in neurons. Saffron and crocetin extended the life span in the G51D fly PD model. Saffron suppressed the rough-eyed phenotype and the dispersion of the size histogram of the ocular long axis in the eye of A30P fly PD model. Saffron had a cytoprotective effect on a human neuronal cell line with α-synuclein fibrils. These data showed that saffron and its constituent crocetin have protective effects on the progression of PD disease in animals in vivo and suggest that saffron and crocetin can be used to treat PD.

Topics: alpha-Synuclein; Animals; Animals, Genetically Modified; Carotenoids; Cell Line; Crocus; Disease Models, Animal; Drosophila melanogaster; Female; Humans; Longevity; Male; Motor Activity; Mutation; Neurons; Neuroprotective Agents; Parkinson Disease; Retinal Degeneration; Vitamin A

The natural carotenoid crocetin has been reported to suppress phenotypes of an experimental myopia model in mice. We investigated the minimum effective dose to prevent myopia progression in a murine model. Three-week-old male mice (C57B6/J) were equipped with a -30 diopter (D) lens to induce myopia, and fed with normal chow, 0.0003%, or 0.001% of crocetin-containing chow. Changes in refractive errors and axial lengths (AL) were evaluated after three weeks. Pharmacokinetics of crocetin in the plasma and the eyeballs of mice was evaluated with specific high sensitivity quantitative analysis using liquid chromatography tandem mass spectrometry (LC-MS/MS) to determine the minimum effective dosage. A concentration of 0.001% of crocetin-containing chow showed a significant (

Topics: Animals; Carotenoids; Dietary Supplements; Disease Models, Animal; Disease Progression; Eye; Male; Mice; Mice, Inbred C57BL; Myopia; Vitamin A

Crocetin (CRT) has shown various neuroprotective effects such as antioxidant activities and the inhibition of amyloid β fibril formation, and thus is a potential therapeutic candidate for Alzheimer's disease (AD). However, poor water solubility and bioavailability are the major obstacles in formulation development and pharmaceutical applications of CRT. In this study, a novel water-soluble CRT-γ-cyclodextrin inclusion complex suitable for intravenous injection was developed. The inclusion complex was nontoxic to normal neuroblastoma cells (N2a cells and SH-SY5Y cells) and AD model cells (7PA2 cells). Furthermore, it showed stronger ability to downregulate the expression of C-terminus fragments and level of amyloid β in 7PA2 cell line as compared to the CRT free drug. Both inclusion complex and CRT were able to prevent SH-SY5Y cell death from H

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Blood-Brain Barrier; Carotenoids; CHO Cells; Cricetulus; Disease Models, Animal; gamma-Cyclodextrins; Humans; Rats; Rats, Sprague-Dawley; Vitamin A

Dengue virus (DENV) infection is one of the most widespread mosquito-borne viral infections. Liver injury is commonly observed in severe DENV infection, and the present study aimed to examine the efficacy of crocetin treatment in an immunocompetent mouse model of DENV infection exhibiting liver injury. The efficacy of crocetin treatment in DENV-induced liver injury was assessed via both transaminase levels and histopathology analysis. A real-time polymerase chain reaction array was then used to describe the expression of 84 apoptosis-related genes. Using real-time RT-PCR and Western blot analysis, the gene expressions of host factors were investigated. Additionally, the effect of crocetin in NF-kB signaling during DENV infection was studied. We did not observe any significant reduction in virus production when DENV-infected mice were treated with crocetin. However, DENV-infected mice treated with crocetin showed reduced DENV-induced apoptosis. The real-time polymerase chain reaction array revealed pro-inflammatory cytokine expressions to be significantly reduced in the crocetin-treated DENV-infected mice. We also found that crocetin could effectively modulate antioxidant status in DENV-infected mice. Moreover, crocetin demonstrated the ability to reduce the nuclear translocation of NF-kB in DENV-infected mice. Our results suggest that crocetin treatment does not inhibit DENV replication in the liver of DENV-infected mice; however, we did find that crocetin improves host responses that reduce liver injury.

Topics: Active Transport, Cell Nucleus; Animals; Apoptosis; Carotenoids; Dengue; Dengue Virus; Disease Models, Animal; Gene Expression; Liver; Liver Diseases; Male; Mice; Mice, Inbred BALB C; Transaminases; Virus Replication; Vitamin A

Topics: Adenosine Triphosphate; Animals; Calcium; Carotenoids; Cell Line, Tumor; Cell Survival; Chromatography, High Pressure Liquid; Crocus; Disease Models, Animal; Flowers; HEK293 Cells; Humans; Light; Mice; Neurodegenerative Diseases; Neuroprotective Agents; Plant Extracts; Rats, Sprague-Dawley; Receptors, Purinergic P2X7; Retina; Retinal Degeneration; Vitamin A

We investigated the effects of oral and ocular administration of crocetin in a murine retinal vein occlusion (RVO) model. Crocetin is a type of carotenoid contained in the fruit of gardenia (. This study was performed on a murine RVO model, which was created by laser irradiation of retinal veins. We evaluated the retinal thickness after the oral administration of crocetin (100 mg/kg) 1 and 6 h before laser irradiation, and immediately, 6 h, 12 h, and 18 h after laser irradiation in the murine RVO model. In addition, we measured the retinal layer thickness after administration of crocetin eye drops (0.03% or 0.10%) immediately, 6 h, and 12 h after laser irradiation. Western blotting of retinal tissue was used to determine the expression levels of matrix metalloproteinase (MMP-9), tumor nuclear factor (TNF-α), and occludin after oral administration of crocetin.. Oral and ocular administration of crocetin improved retinal edema in the murine RVO model. Crocetin administration statistically significantly suppressed overexpression of MMP-9 and TNF-α, and reversed the reduction of occludin.. These findings indicate that crocetin can protect retinal tight junctions by suppressing retinal edema through an anti-inflammatory effect, which suggests that crocetin may be useful for RVO disease.

Topics: Administration, Ophthalmic; Administration, Oral; Animals; Carotenoids; Disease Models, Animal; Edema; Inflammation Mediators; Male; Matrix Metalloproteinase 9; Mice; Occludin; Ophthalmic Solutions; Retina; Retinal Vein Occlusion; Tight Junctions; Tumor Necrosis Factor-alpha; Vitamin A

The study was designed to determine the safety and pharmacokinetics of intraocular crocetin and examine whether crocetin inhibits the development of proliferative vitreoretinopathy (PVR) in a rabbit model. In the toxicity study, the right eyes of rabbits were injected with 0.2 μmol or 0.4 μmol crocetin. The left eyes were injected with 0.1 ml phosphate buffered saline (PBS) containing the same concentration of DMSO. Fundus photography, optical coherence tomography (OCT), and electroretinogram (ERG) were obtained at baseline and 14 days. Afterward, the eyes were enucleated for histopathological analysis and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) assay. In the pharmacokinetic study, the eyes received an intravitreous injection of 0.4 μmol crocetin to detect vitreous drug levels with HPLC at specific time points. In the efficacy study, PVR was induced with an intravitreal injection of ARPE-19 cells in rabbits. Then ten eyes were injected with 0.4 μmol crocetin, and the other 10 eyes received 0.1 ml PBS. Fundus photography, OCT and ERG were performed at days 3 and 7 and weekly for a total of 4 weeks after injection. Afterward, the eyes were enucleated and subjected to histological analysis and TUNEL staining. The results demonstrated no signs of retinal toxicity. Intravitreal injection of 0.4 μmol crocetin had a half-life of 4.231 h. Treatment with crocetin significantly inhibited the progression of PVR in parallel with a reduced expression of α-SMA, collagen fibers and Ki67. These results indicate that crocetin is an effective and safe inhibitor of PVR in rabbit models.

Topics: Animals; Carotenoids; Cell Line; Disease Models, Animal; Electroretinography; Humans; Intravitreal Injections; Rabbits; Retina; Vitamin A; Vitreoretinopathy, Proliferative

Increased global incidence of myopia necessitates establishment of therapeutic approaches against its progression. To explore agents which may control myopia, we screened 207 types of natural compounds and chemical reagents based on an activity of a myopia suppressive factor, early growth response protein 1 (Egr-1) in vitro. Among the candidates, crocetin showed the highest and dose-dependent activation of Egr-1. For in vivo analysis, experimental myopia was induced in 3-week-old C57BL/6 J mice with -30 diopter (D) lenses for 3 weeks. Animals were fed with normal or mixed chow containing 0.003% (n = 19) and 0.03% (n = 7) of crocetin during myopia induction. Refraction and axial length were measured at 3-week-old and the 6-week-old with an infrared photorefractor and a SD-OCT system. Compared to controls (n = 14), crocetin administration showed a significant smaller change of refractive errors (-13.62 ± 8.14 vs +0.82 ± 5.81 D for 0.003%, p < 0.01, -2.00 ± 4.52 D for 0.03%, p < 0.01) and axial elongation (0.27 ± 0.03 vs 0.22 ± 0.04 mm for 0.003%, p < 0.01, 0.23 ± 0.05 mm for 0.03%, p < 0.05). These results suggest that a dietary factor crocetin may have a preventive effect against myopia progression.

Topics: Administration, Oral; Animals; Carotenoids; Disease Models, Animal; Early Growth Response Protein 1; Mice; Mice, Inbred C57BL; Myopia; Refraction, Ocular; Refractive Errors; Vitamin A

Trans sodium crocetinate (TSC) has been reported to exert a protective effect against cerebral ischemia/reperfusion (I/R) injury. However, whether TSC protects against myocardial ischemia/reperfusion (MI/R) injury remains unknown. Herein, we found that TSC treatment reduced myocardial infract size and elevated serum LDH and CK activities of MI/R rats. TSC administration attenuated oxidative stress in MI/R rats and H9C2 cells exposed to oxygen glucose deprivation/reperfusion (OGD/R). TSC administration relieved I/R-induced myocardial apoptosis in vivo and in vitro, as evidenced by reduced number of TUNEL positive cells, accompanying with marked decreases in caspase-3 activity and Bax protein level and an increase in Bcl-2 protein level. TSC treatment markedly increased SIRT3 activity and SIRT3 and SOD2 protein levels, and could also diminished the phosphorylation of FOXO3a protein. Additionally, TSC treatment attenuated the acetylation of FOXO3a and SOD2 protein. But, these effects were obviously blocked by SIRT3 knockdown. Besides, SIRT3 knockdown blocked the cardioprotective effect of TSC on OGD/R-induced oxidative stress, apoptosis and mitochondrial dysfunction in vitro. In summary, TSC alleviates I/R-induced myocardial oxidative stress and apoptosis via the SIRT3/FOXO3a/SOD2 signaling pathway. Our study suggests that TSC may become a novel drug for the treatment of MI/R injury.

Topics: Animals; Antioxidants; Apoptosis; Carotenoids; Coronary Occlusion; Disease Models, Animal; Forkhead Box Protein O3; Humans; Male; Myocardial Infarction; Myocardium; Oxidative Stress; Rats; Rats, Wistar; Reperfusion Injury; RNA, Small Interfering; Signal Transduction; Sirtuin 3; Vitamin A

Crocetin, an active ingredient of saffron, has been recognized as a potent antioxidant. Plant extracts or their components may be useful in ameliorating the various diseases, including neurodegenerative disorders. This study investigated the effects of crocetin on oxidative damage induced by chronic restraint stress in the rat brain. For this reason, rats were kept in the restrainers for 1 hour every day, for 21 consecutive days. The animals were injected crocetin (20, 40, 60 mg/kg) or vehicle daily for 21 days. Findings showed that the immobility time significantly increased in the rodents subjected to the chronic stress compared with the normal group. However, the number of crossing beams in the rats submitted to the chronic stress significantly decreased versus the non-stress rats. Treatment with crocetin ameliorated the immobility time and the number of crossing in the chronic restraint stress rats versus the non-treated stress group. Crocetin also reverted the levels of MDA and GSH and also the activities of antioxidant enzymes to the normal levels in the stress groups.. The present study suggests that crocetin may be useful for the management of depressantlike effects induced by chronic stress through decreasing oxidative damage in the brain.

Topics: Animals; Antioxidants; Carotenoids; Corticosterone; Depression; Disease Models, Animal; Exploratory Behavior; Glutathione; Glutathione Reductase; Immobilization; Male; Malondialdehyde; Random Allocation; Rats; Rats, Wistar; Swimming; Vitamin A

Polycystic ovarian syndrome (PCOS), the most common endocrine disorder of women in reproductive age, is typical with hyperandrogenism and disturbance of the hypothalamus-pituitary-ovary (HPO) axis, i.e. abnormal expression of hypothalamic gonadotropin-releasing hormone (GnRH) followed by the elevated ratio of serum luteinizing hormone (LH) level to follicle-stimulating hormone (FSH) level. This derangement might have a close relationship with hypothalamic kisspeptin expression that is thought to be a key regulator of GnRH. Crocetin, one of the main components of Saffron clinically used as traditional medicine in gynecology diseases, was evaluated for its therapeutic effects on PCOS induced by prenatally exposure to dihydrotestosterone (DHT) in mice. Herein, we found that DHT-treated mice showed a similar phenotype to human PCOS such as heavier ovary, prolonged diestrus, multiple enlarged follicles with fewer corpus luteum, and higher LH and testosterone levels. Kisspeptin expression was lower in anteroventral periventricular nucleus (AVPV) but higher in arcuate nucleus (ARC). Treatment of crocetin prevented the prolongation of diestrus and reduction in corpora luteum, recover the levels of GnRH, FSH, LH, progesterone (P4), estradiol (E2) and testosterone (T), and increase the kisspeptin level in AVPV but reduce that in ARC. The present study provides in vivo evidence that crocetin improved the PCOS in mice via increasing AVPV-kisspeptin and reducing ARC-kisspeptin expression.

Topics: Animals; Arcuate Nucleus of Hypothalamus; Carotenoids; Crocus; Dihydrotestosterone; Disease Models, Animal; Drugs, Chinese Herbal; Estrous Cycle; Female; Hypothalamo-Hypophyseal System; Hypothalamus, Anterior; Kisspeptins; Neurons; Ovary; Polycystic Ovary Syndrome; Vitamin A

Given study evaluates the cardioprotective effect of crocetin in myocardial infracted (MI) rats. MI was produced by administering isoproterenol (90mg/kg/day, i.p.) in rats for two consecutive days. all the animals were divided in to four groups such as control group receives only saline; MI group which receives only isoproterenol and crocetin treated group which receives crocetin (50, 100 and 200mg/kg/day, p.o.) for the duration of 15 days. At the end of dosing left ventricular functions was assessed to estimate its effect on cardiac functions. Catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), creatine kinase (CK-MB), lactate dehydrogenase (LDH) and inflammatory cytokines were determined in the cardiac tissue homogenate. Histopathology study was also carried out using hematoxylin and eosin staining. Immunohistochemistry was done for the estimation of Caspase-3, Bcl-2, Bax and Nrf-2 level in the myocardial tissues of MI rats. Result of the study suggested that GSH, CAT, CK-MB, and LDH were (p<0.01) increased in the tissue homogenate of crocetin treated group than MI group. However crocetin significantly (p<0.01) decreases the level of MDA and activity of SOD in the tissue homogenate than MI group. It was observed that treatment with crocetin attenuates the level of inflammatory cytokines in the myocardial tissues of MI rats. Moreover level of caspase-3, Bax and Nrf-2 significantly reduced and Bcl-2 enhanced in the myocardial tissues of MI rats than MI group. The altered cellular architecture of heart tissue sections in the myocardial infracted rats were reversed by administration of crocetin treatment. Taking all these data together, it may be suggested that the crocetin act as a possible protective agent in myocardial infarction by decreasing oxidative stress and inflammatory cytokines and thereby attenuates the apoptosis of myocardial cells.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Biomarkers; Cardiotonic Agents; Carotenoids; Caspase 3; Cytokines; Disease Models, Animal; Hemodynamics; Inflammation Mediators; Isoproterenol; Male; Myocardial Infarction; Myocardium; NF-E2-Related Factor 2; Oxidative Stress; Rats, Wistar; Ventricular Function, Left; Vitamin A

Crocetin is an ingredient of traditional Chinese medicine and has therapeutic potential in various diseases due to its pharmacological properties, such as neuroprotection, anti-oxidative stress, and anti-inflammation. These properties might benefit the treatment of spinal cord injury. In the present study, we tested the effect of crocetin on neurite growth and sensorimotor dysfunction in a rat model of spinal cord injury. We evaluated the viability of cultured hippocampal neurons with tetrazolium dye and lactate dehydrogenase assays, visualized neurites and axons with antibody staining, and monitored motor and sensorimotor functions in rats with spinal cord injury using the Basso, Beattie, and Bresnahan assay and the contact plantar placement test, respectively, and measured cytokine expression using enzyme-linked immuno-absorbent assays. We found that crocetin (1) did not alter the viability of cultured hippocampal neurons; (2) accelerated neurite growth with preference for the longest process in individual hippocampal neurons; (3) reversed the inhibition of neurite growth by chondroitin sulfate proteoglycan and NogoA; (4) facilitated the recovery of motor and sensorimotor functions after spinal cord injury; and (5) did not inhibit pro-inflammatory responses, but restored the innervation of the descending 5-HT system in injured spinal cord. Crocetin promotes neurite growth and facilitates the recovery of motor and sensorimotor functions after spinal cord injury, likely through repairing neuronal connections.

Topics: Animals; Antioxidants; Behavior, Animal; Carotenoids; Disease Models, Animal; Hippocampus; Inflammation; Motor Activity; Neurites; Neurons; Rats; Recovery of Function; Spinal Cord Injuries; Vitamin A

Crocetin is the main component of saffron and exhibits anti-oxidative and anti-inflammatory effects. Neuroinflammation and oxidative stress have been recognized to play a crucial role in the pathogenesis of neuropathic pain. We investigated the effect of crocetin in a mouse model with neuropathic pain induced by spared nerve injury (SNI). Crocetin was intrathecally perfused at various doses for up to 12 days starting 3 days before the surgery. Behavioral tests were performed to determine pain sensitivity. The concentrations of proinflammatory cytokines tumor necrosis factor (TNF-α) and interleukin-1β (IL-1β) were measured to assess neuroinflammation. In addition, the enzymatic activity of superoxide dismutase (SOD) was measured to reveal the oxidative stress level. We found that repeated treatment with crocetin dose-dependently attenuated mechanical and thermal allodynia in SNI mice. In addition, treatment with high dose of crocetin reduced SNI-induced increase of TNF-α and IL-1β. Crocetin also restored the activity of mitochondrial MnSOD which was reduced in the sciatic nerve and the spinal cord of SNI mice. Collectively, our data demonstrate that crocetin effectively attenuates the neuropathic pain and significantly suppresses oxidative stress and neuroinflammation in the SNI mouse model, supporting the potential of crocetin in the treatment against neuropathic pain.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Biomarkers; Carotenoids; Crocus; Disease Models, Animal; Dose-Response Relationship, Drug; Inflammation Mediators; Interleukin-1beta; Male; Mice, Inbred Strains; Neuralgia; Oxidative Stress; Peripheral Nerve Injuries; Phytotherapy; Sciatic Nerve; Spinal Cord; Superoxide Dismutase; Tumor Necrosis Factor-alpha; Vitamin A

Trans-sodium crocetinate (TSC) is a novel synthetic carotenoid compound that improves diffusion of small molecules, including oxygen, in solutions. TSC provides neuroprotection in healthy rats and rabbits. This study seeks to determine whether TSC is neuroprotective in obese mice. Sixteen-week-old CD-1 male mice that had been fed a high-fat diet for 10 weeks were subjected to a 90-min middle cerebral arterial occlusion (MCAO). They received TSC by two boluses through a tail vein 10 min after the onset of MCAO and reperfusion, respectively, with doses of 0.14, 0.28, and 0.7 mg/kg or by a bolus-infusion-bolus strategy with a dose of 0.14 mg/kg during MCAO. The neurological outcome was evaluated 72 hr after MCAO. Brain tissues were harvested 24 hr after MCAO to measure nitrotyrosine-containing proteins, 4-hydroxy-2-nonenal, matrix metalloproteinase (MMP)-2 and -9 activity and expression, and inflammatory cytokines. TSC given in the two-bolus strategy did not improve the neurological outcome. The bolus-infusion-bolus strategy significantly reduced brain edema, infarct volume, and hemorrhagic transformation and improved neurological functions. TSC reduced nitrotyrosine-containing proteins, MMP-9 activity and expression, and inflammatory cytokines in ischemic brain tissues. Our results indicate that TSC delivered by the bolus-infusion-bolus strategy provides neuroprotection in obese mice. This protection may occur through reduction of oxidative stress, MMP-9 activity, or inflammatory cytokines in the ischemic brain tissues.

Topics: Aldehydes; Analysis of Variance; Animals; Brain; Brain Ischemia; Carotenoids; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Male; Matrix Metalloproteinases; Mice; Nervous System Diseases; Neuroprotective Agents; Obesity; Oxidative Stress; Reperfusion; Tyrosine; Vitamin A

Endothelial dysfunction, characterized by an enhancement in vasoconstriction, is clearly associated with hypertension. Saffron (Crocus sativus L.) bioactive compounds have been recognized to have hypotensive properties. Recently, we have reported that crocetin exhibits potent vasodilator effects on isolated aortic rings from hypertensive rats. In this work, we have aimed to analyze the anticontractile ability of crocetin or crocetin esters pool (crocins) isolated from saffron. Thus, we have studied the effects of saffron carotenoids on endothelium-dependent and -independent regulation of smooth muscle contractility in genetic hypertension.. We have measured the isometric responses of aortic segments with or without endothelium obtained from spontaneously hypertensive rats. The effects of carotenoids were studied by assessing the endothelial modulation of phenylephrine-induced contractions (10(-9)-10(-5) M) in the presence or absence of crocetin or crocins. The role of nitric oxide and prostanoids was analyzed by performing the experiments with L-NAME (NG-nitro-l-arginine methyl ester) or indomethacin (both 10(-5) M), respectively.. Crocetin, and to a minor extent crocins, diminished the maximum contractility of phenylephrine in intact rings, while crocins, but not crocetin, increased this contractility in de-endothelizated vessels. In the intact vessels, the effect of crocetin on contractility was unaffected by indomethacin but was abolished by L-NAME. However, crocetin but not crocins, lowered the already increased contractility caused by L-NAME.. Saffron compounds, but especially crocetin have endothelium-dependent prorelaxing actions. Crocins have procontractile actions that take place via smooth muscle cell mechanisms. These results suggest that crocetin and crocins activate different mechanisms involved in the vasoconstriction pathway in hypertension.

Topics: Animals; Aorta; Carotenoids; Crocus; Disease Models, Animal; Esters; Hypertension; Male; Muscle, Smooth, Vascular; Plant Extracts; Rats; Rats, Inbred SHR; Vitamin A

Trans-sodium crocetinate (TSC) is a novel carotenoid compound capable of enhancing the diffusion of small molecules in aqueous solutions. TSC improves the diffusion of oxygen and glucose, and increases oxygenation in ischemic brain tissue. TSC also dampens the intensity of an ischemic challenge during an ongoing ischemic event. The current study examined the impact of TSC in rat models of ischemic and hemorrhagic stroke. Rat three vessel occlusion (3VO), and combined 3VO and one vessel occlusion (3VO/1VO) models of ischemic stroke were evaluated for structural and behavioral outcomes. The effects of TSC were also tested in a rat model of intracerebral hemorrhage (ICH). Delayed treatment with TSC reduced infarct volume in a rodent model of transient focal ischemia involving either 2 or 6h of ischemia. Neurological outcomes, based on a multi-scale assessment and automated gait analysis, also were improved by TSC treatment. Additionally, TSC reduced edema and hemorrhagic volume in a rat model of ICH. An optimal therapeutic candidate for early intervention in ischemic stroke should be effective when administered on a delayed basis and should not aggravate outcomes associated with hemorrhagic stroke. The current findings demonstrate that delayed TSC treatment improves outcomes in experimental models of both ischemic and hemorrhagic stroke. Together, these findings suggest that TSC may be a safe and beneficial therapeutic modality for early stroke intervention, irrespective of the type of stroke involved.

Topics: Animals; Biomechanical Phenomena; Brain; Brain Edema; Brain Ischemia; Carotenoids; Cerebral Hemorrhage; Disease Models, Animal; Gait; Male; Neuroprotective Agents; Pattern Recognition, Automated; Random Allocation; Rats, Sprague-Dawley; Severity of Illness Index; Stroke; Treatment Outcome; Vitamin A

Crocetin is one of the major active constituents of saffron extract, and it is a carotenoid compound that prevents reactive oxygen species and has anti-inflammation and anti-apoptosis characteristics. This study aims to investigate whether crocetin repairs myocardial damage in vivo after ischemia reperfusion (I/R) and the mechanisms underlying its cardioprotective effects. Male Wistar rats were randomly allocated into three groups: Sham, I/R, CRO. Crocetin (50mg/kg/day, i.g.) or sodium carboxymethylcellulose (CMC-Na) was intragastrically administered to Wistar rats for 7 days before operation. Myocardial ischemia reperfusion injury (MIRI) was induced by occluding the left anterior descending (LAD) coronary artery for 45min and subsequent reperfusion for 3h. The cardioprotective effects of crocetin were evaluated by biochemical values, histopathological observations and the antiapoptotic relative proteins and gene expressions. In the rat model, pretreatment with 50mg/kg crocetin reduced the cardiac injury, oxidative stress and inflammation compared with that of the non-treated rats, as shown by the decreased levels of infarct size, creatine kinase-MB (CK-MB), malondialdehyde (MDA), and tumor necrosis factor-alpha (TNF-α) activity and the increased levels of total superoxide dismutase (T-SOD) and inflammation cytokines interleukin-10 (IL-10) activity. Crocetin activation also decreases the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining positive percentage and Bax expression, elevated Bcl-2 and endothelial nitric oxide synthase (eNOS) expression and nitrite (NO) production, these indicating that crocetin can suppress the apoptosis damage. These results indicate that crocetin can provide protection against MIRI in rats by inhibiting ROS production, blocking inflammation, and reducing myocardium apoptosis.

Topics: Animals; Cardiotonic Agents; Carotenoids; Disease Models, Animal; Male; Myocardial Reperfusion Injury; Random Allocation; Rats; Rats, Wistar; Treatment Outcome; Vitamin A

Hypertension is associated with endothelial dysfunction characterized by decreased vasorelaxation. Crocetin, a bioactive compound of saffron, exhibits favorable cardiovascular properties. We analyze the vasomodulatory effects of crocetin in hypertension.. Myographical experiments were performed to compare the relaxation induced by acetylcholine (ACH) on aortic rings from normotensive (Wistar) and hypertensive (SHR) rats, incubated with or without crocetin or saffron extract and L-NAME or indomethacin. Extracts were also assayed in deendothelialized rings. UV-vis spectrophotometry and HPLC-DAD were used to characterize and quantify the saffron used.. Crocetin enhanced the ACH relaxations in aorta from hypertensive (strongly) and normotensive rats (weakly). Saffron extract did not modify this. Crocetin plus L-NAME abolished the relaxant response in SHR but not in Wistar aorta. Crocetin plus indomethacin did not modify the indomethacin response in either SHR or Wistar aorta. Crocetin in rubbed segments did not modify the ACH responses. In contrast, saffron increased this response in rubbed segments from SHR but not Wistar rats.. Crocetin exerts healthy vasomodulatory effects in hypertension, strongly improving endothelium-dependent ACH relaxations via endothelial nitric oxide but not the cyclooxygenase pathway. This work proposes that crocetin supplements are a possible complement in the therapy of hypertension.

Topics: Acetylcholine; Animals; Antihypertensive Agents; Carotenoids; Crocus; Cyclooxygenase Inhibitors; Disease Models, Animal; Dose-Response Relationship, Drug; Hypertension; Male; Myography; Nitric Oxide; Nitric Oxide Synthase; Phytotherapy; Plants, Medicinal; Rats, Inbred SHR; Rats, Wistar; Vasodilation; Vasodilator Agents; Vitamin A

Chronic cerebral hypoperfusion produces brain ischemia and is associated with related memory impairment. In this study, we prepared crocetin from crocin (the main water soluble pigment in saffron stigmas) in an acidic hydrolysis and its purity was evaluated using spectrophotometry, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The memory enhancing effect of crocetin was then studied in a rat model of chronic cerebral hypoperfusion. Different doses of crocetin were administered intraperitoneally (i.p.). The spatial learning and memory function were tested using Morris water maze, after permanent occlusion of common carotids. Finally, the animals were euthanatized in CO2 chamber and histopathological changes in cerebral cortex and hippocampus were investigated. The results indicated that the escape latency time significantly decreased in crocetin treated rats, in comparison with control animals. Also, the percentage of time spent and traveled distance in target quadrant on final test trial day increased in crocetin 8 mg/kg group, compared to control group, while no difference was observed between groups in swimming speed. All behavioral results were confirmed by histopathological analysis. According to our data, crocetin (8 mg/kg) could properly protect cerebrocortical and hippocampus neurons against ischemia. In conclusion, treatment with crocetin could effectively prevent neuropathological alterations in hippocampus thereby resulting in improvement of spatial learning memory in rats after chronic cerebral hypoperfusion.

Topics: Animals; Carotenoids; Dementia, Vascular; Disease Models, Animal; Hippocampus; Male; Maze Learning; Neuroprotective Agents; Rats; Rats, Wistar; Vitamin A

Ischemic neuronal damage is a common feature of occlusive strokes, hemorrhagic strokes, and traumatic brain injury. In addition, ischemia can be an anticipated or unanticipated complication of a variety of surgical procedures. Most therapeutic strategies for managing ischemic injury seek to re-establish blood flow, suppress neural metabolism, and/or limit specific cellular injury cascades. An alternative therapeutic approach is to enhance the delivery of metabolic substrates to ischemic tissue. This strategy is typified by efforts to increase tissue oxygenation by elevating the levels of circulating oxygen. Our studies are examining a complementary approach in which the delivery of metabolic substrates is enhanced by facilitating the diffusion of oxygen and glucose from the vasculature into neural tissue during ischemia. This is achieved by increasing the diffusivity of small molecules in aqueous solutions, such as plasma and interstitial fluid. The carotenoid compound, trans-sodium crocetinate (TSC) is capable of increasing oxygen and glucose diffusivity, and our studies demonstrate that TSC increases cerebral tissue oxygenation in the penumbra of a focal ischemic event. In addition, TSC treatment reduces the volume of cerebral infarction in rodent models of both permanent and temporary focal ischemia. This strategy of "metabolic reflow" thus blunts the metabolic challenge in partially-perfused tissue and reduces ischemic neural injury.

Topics: Animals; Carotenoids; Cerebral Infarction; Cerebrovascular Circulation; Disease Models, Animal; Glucose; Infarction, Middle Cerebral Artery; Male; Oxygen; Oxygen Consumption; Rats; Rats, Sprague-Dawley; Time Factors; Vitamin A

Hemorrhagic shock results in cellular damage and cell death. A primary mechanism is cellular apoptosis from mitochondrial damage. This study demonstrated that administration of crocetin to experimental animals during resuscitation from shock significantly improved postshock survival and reduced apoptosis. Crocetin is a component of saffron and has long been used in traditional medicine in Asia.. Male Sprague-Dawley rats (350 ± 30 g) were randomly assigned to 1 of 4 groups of 8 animals. Hemorrhagic shock was induced by withdrawing blood until the mean arterial pressure was 35-40 mm Hg, and blood pressure was maintained at that level for 60 minutes with further withdrawals as needed. Resuscitation was carried out by administration of 21 mL/kg lactated Ringer's solution and return of shed blood, with or without concurrent administration of crocetin (2 mg/kg). Control animals were sham-treated with surgical preparation, without shock or resuscitation, and with and without crocetin. Rats were sacrificed 24 hours after completion of resuscitation. The extent of activation of hepatic apoptosis was established by measuring levels of hepatic cytosolic cytochrome c, caspase-3, and bcl-2. A separate group of 53 animals treated identically was used to assess survival.. Crocetin administration during resuscitation resulted in less extensive activation of hepatic apoptosis and significantly increased survival relative to controls.. Crocetin administration to experimental animals during resuscitation post hemorrhage increased survival, at least in part by protecting the liver from activation of apoptotic cell death. This agent continues to show promise as a potential treatment strategy for hemorrhagic shock.

Topics: Animals; Apoptosis; Carotenoids; Caspase 3; Crocus; Cytochromes c; Disease Models, Animal; Liver; Male; Phytotherapy; Plant Extracts; Proto-Oncogene Proteins c-bcl-2; Random Allocation; Rats; Rats, Sprague-Dawley; Resuscitation; Shock, Hemorrhagic; Vitamin A

Trans sodium crocetinate (TSC) is a synthetic small-molecule antioxidant that has the ability to enhance oxygen diffusion to hypoxic tissue. Because TSC is a promising drug candidate to treat acute ischemic stroke (AIS), we tested the hypothesis that TSC may be neuroprotective following cerebral ischemia using a rabbit small clot embolic stroke model (RSCEM) using clinical rating scores as the endpoint. TSC or saline was administered IV following the injection of small blood clots into the brain vasculature. Behavior was measured 24 h following embolization in order to calculate the effective stroke dose (P(50)) that produces neurological deficits in 50% of the rabbits. A treatment is considered beneficial if it significantly increases the P(50) compared to control. TSC (0.25 mg/kg) given 5 or 60 min following embolization significantly (p<0.05) increased P(50) values by 104% and 181%; but not when given 3 h post-embolization (48% increase, p>0.05). tPA (3.3 mg/kg) produced a significant increase in P(50) when given 1, but not 3 h following embolization. In combination studies, when TSC was administered 1 h and tPA was given either 1 or 3 h following embolization, the group P(50) values were increased by 291% and 140%, respectively. In addition, TSC plus tPA administered 3 h following embolization significantly (p<0.05) increased the group P(50) value by 90%. There were no significant effects (p>0.05) of either TSC alone or TSC administered in combination with tPA on intracerebral hemorrhage incidence. This study suggests that TSC may be used for the treatment of AIS either alone or when administered before or concomitant with tPA to improve clinical rating scores with a therapeutic window for TSC therapy up to 3 h in rabbits. Moreover, it appears that TSC can be administered with tPA, since the combination did not result in any significant change in intracerebral hemorrhage incidence.

Topics: Animals; Brain Infarction; Brain Ischemia; Carotenoids; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Administration Schedule; Drug Synergism; Drug Therapy, Combination; Drug-Related Side Effects and Adverse Reactions; Emergency Medical Services; Fibrinolytic Agents; Intracranial Embolism; Male; Neuroprotective Agents; Rabbits; Time Factors; Tissue Plasminogen Activator; Treatment Outcome; Vitamin A

Glioblastoma (GB) tumors typically exhibit regions of hypoxia. Hypoxic areas within the tumor can make tumor cells less sensitive to chemotherapy and radiation therapy. Trans-sodium crocetinate (TSC) has been shown to transiently increase oxygen to hypoxic brain tumors. The authors examined whether this improvement in intratumor oxygenation translates to a therapeutic advantage when delivering standard adjuvant treatment to GBs.. The authors used C6 glioma cells to create a hypoxic GB model. The C6 glioma cells were stereotactically injected into the rat brain to create a tumor. Fifteen days later, MR imaging was used to confirm the presence of a glioma. The animals were randomly assigned to 1 of 3 groups: 1) temozolomide alone (350 mg/m(2)/day for 5 days); 2) temozolomide and radiation therapy (8 Gy); or 3) TSC (100 microg/kg for 5 days), temozolomide, and radiation therapy. Animals were followed through survival studies, and tumor response was assessed on serial MR images obtained at 15-day intervals during a 2-month period.. Mean survival (+/- SEM) of the temozolomide-alone and the temozolomide/radiotherapy groups was 23.2 +/- 0.9 and 29.4 +/- 4.4 days, respectively. Mean survival in the TSC/temozolomide/radiotherapy group was 39.8 +/- 6 days, a statistically significant improvement compared with either of the other groups (p < 0.05). Although tumor size was statistically equivalent in all groups at the time of treatment initiation, the addition of TSC to temozolomide and radiotherapy resulted in a statistically significant reduction in the MR imaging-documented mean tumor size at 30 days after tumor implantation. The mean tumor size in the TSC/temozolomide/radiotherapy group was 18.9 +/- 6.6 mm(2) compared with 42.1 +/- 2.7 mm(2) in the temozolomide-alone group (p = 0.047) and 35.8 +/- 5.1 mm(2) in the temozolomide/radiation group (p = 0.004).. In a hypoxic GB model, TSC improves the radiological and clinical effectiveness of temozolomide and radiation therapy. Further investigation of this oxygen diffusion enhancer as a radiosensitizer for hypoxic brain tumors seems warranted.

Topics: Animals; Antineoplastic Agents, Alkylating; Brain Neoplasms; Carotenoids; Cell Line, Tumor; Combined Modality Therapy; Dacarbazine; Diffusion; Disease Models, Animal; Glioblastoma; Hypoxia, Brain; Kaplan-Meier Estimate; Magnetic Resonance Imaging; Neoplasm Transplantation; Oxygen; Radiation-Sensitizing Agents; Rats; Rats, Sprague-Dawley; Temozolomide; Vitamin A

The amelioration of insulin resistance by treatment with crocetin is closely related to the hypolipidaemic effect. The present study is designed to clarify the insulin-sensitizing mechanism of crocetin by elucidating the mechanism of regulation of lipid metabolism by crocetin.. Rats given a high-fat diet were treated with crocetin for 6 weeks before hyperinsulinaemic-euglycaemic clamp. 14C-palmitate was used as tracer to track the fate of non-esterified fatty acids or as substrate to measure beta-oxidation rate. Triglyceride clearance in plasma and lipoprotein lipase activity in tissues were tested. Content of lipids in plasma and tissues was determined. Real-time PCR was used to assay the level of mRNA from genes involved in non-esterified fatty acid and triglyceride uptake and oxidation.. Crocetin prevented high-fat-diet induced insulin resistance (increased clamp glucose infusion rate), raised hepatic non-esterified fatty acid uptake and oxidation, accelerated triglyceride clearance in plasma, enhanced lipoprotein lipase activity in liver, and reduced the accumulation of detrimental lipids (DAG and long-chain acyl CoA) in liver and muscle. Genes involved in hepatic lipid metabolism which are regulated by peroxisome proliferator-activated receptor-alpha, were modulated to accelerate lipid uptake and oxidation.. Through regulating genes involved in lipid metabolism, crocetin accelerated hepatic uptake and oxidation of non-esterified fatty acid and triglyceride, and reduced lipid availability to muscle, thus decreasing lipid accumulation in muscle and liver, and consequently improving sensitivity to insulin.

Topics: Adipose Tissue; Animals; Blood Glucose; Carnitine O-Palmitoyltransferase; Carotenoids; Dietary Fats; Disease Models, Animal; Dyslipidemias; Fatty Acids, Nonesterified; Glucose Clamp Technique; Hypolipidemic Agents; Insulin; Insulin Resistance; Lipase; Lipoprotein Lipase; Lipoproteins; Liver; Male; Muscle, Skeletal; Oxidation-Reduction; Polymerase Chain Reaction; Rats; Rats, Wistar; RNA, Messenger; Time Factors; Triglycerides; Vitamin A

Antioxidants have been expected to have potential as antiatherogenic agents. Crocetin is a natural carotenoid antioxidant isolated from Gardenia jasminoids Ellis. Therefore, in the present study, we investigated the inhibitory effect of Crocetin on experimental atherosclerosis in quails. The atherosclerosis model was established by feeding hyperlipidamic diet to quail and Crocetin (25, 50, 100 mg/kg/day) was administered by oral gavage. At the 9th week, serum lipids, malondialdehyde and nitric oxide were measured, and Hematoxylin-Eosin (H&E) stains was used to investigate the histopathological changes of aorta. Results showed that Crocetin could reduce the levels of serum total cholesterol, triglyceride, low density lipoprotein cholesterol and inhibit the formation of aortic plaque. Crocetin could also reduce malondialdehyde and inhibit the descending of nitric oxide in serum. The results suggested that Crocetin could inhibit the formation of atherosclerosis in quails, which might be related to the hypolipidemic effects along with the antioxidative properties of Crocetin.

Topics: Animals; Antioxidants; Aorta; Atherosclerosis; Carotenoids; Cholesterol; Cholesterol, Dietary; Cholesterol, LDL; Cholesterol, VLDL; Diet, Atherogenic; Dietary Fats; Disease Models, Animal; Dose-Response Relationship, Drug; Lipids; Male; Malondialdehyde; Molecular Structure; Nitric Oxide; Quail; Triglycerides; Vitamin A

Crocus sativus L. (saffron) has been used as a spice for flavoring and coloring food preparations, and in Chinese traditional medicine as an anodyne or tranquilizer. Our previous study demonstrated that crocin, a carotenoid pigment of saffron, can suppress the serum deprivation-induced death of PC12 cells by increasing glutathione (GSH) synthesis and thus inhibiting neutral sphingomyelinase (nSMase) activity and ceramide formation. The carotenoid pigments of saffron consist of crocetin di-(beta-d-glucosyl)-ester [dicrocin], crocetin-(beta-d-gentiobiosyl)-(beta-d-glucosyl)-ester [tricrocin] and crocetin-di-(beta-d-gentiobiosyl)-ester [crocin]. Saffron also contains picrocrocin, the substance causing saffron's bitter taste. In this study, to confirm whether neuroprotective effects of saffron are caused solely by crocin, we examined the antioxidant and GSH-synthetic activities of these crocins in PC12 cells under serum-free and hypoxic conditions. Measurements of cell viability, peroxidized membrane lipids and caspase-3 activity showed that the rank order of the neuroprotective potency at a concentration of 10 muM was crocin>tricrocin>dicrocin and picrocrocin (the latter two crocins had a little or no potency). In addition, we show that among these saffron's constituents, crocin most effectively promotes mRNA expression of gamma-glutamylcysteinyl synthase (gamma-GCS), which contributes to GSH synthesis as the rate-limiting enzyme, and that the carotenoid can significantly reduce infarcted areas caused by occlusion of the middle cerebral artery (MCA) in mice.

Topics: Animals; Antioxidants; Brain Infarction; Carotenoids; Caspase 3; Cell Hypoxia; Cell Survival; Crocus; Cyclohexenes; Disease Models, Animal; Glucosides; Glutamate-Cysteine Ligase; Glutathione; Infarction, Middle Cerebral Artery; Lipid Peroxidation; Male; Membrane Lipids; Mice; Molecular Structure; Neurons; Neuroprotective Agents; PC12 Cells; Rats; Structure-Activity Relationship; Terpenes; Time Factors; Vitamin A

Reactive oxygen species (ROS) are implicated as the leading biochemical cause of neuronal death in various neurologic disorders, including Parkinson's disease. In the present study, neuromodulatory effects of crocetin (active constituent of Crocus sativus) in a 6-hydroxydopamine (6-OHDA) model of rat Parkinsonism were investigated. Male Wistar rats were pre-treated with crocetin (25, 50 and 75 microg/kg body weight) for 7 days and subjected to unilateral intrastriatal injection of 10 microg 6-OHDA on day 8. Locomotion and rotation were observed on day 23 post-injection, and after 4 weeks, striatum and substantia nigra were dissected out by decapitation. Activity of antioxidant enzymes and content of dopamine (DA) and its metabolites were estimated in striatum, whereas glutathione (GSH) content and thiobarbituric acid reactive substance (TBARS) were evaluated in substantia nigra. Levels of GSH and dopamine were protected, while TBARS content was attenuated in crocetin-treated groups. The activity of antioxidant enzymes was decreased in the lesion group, but protected in the crocetin-treated groups. These findings were supported by the histopathologic findings in the substantia nigra that showed that crocetin protects neurons from deleterious effects of 6-OHDA. This study revealed that crocetin, which is an important ingredient of diet in India and also used in various systems of indigenous medicine, is helpful in preventing Parkinsonism and has therapeutic potential in combating this devastating neurologic disorder.

Topics: 3,4-Dihydroxyphenylacetic Acid; Analysis of Variance; Animals; Behavior, Animal; Carotenoids; Catalase; Corpus Striatum; Disease Models, Animal; Dopamine; Dose-Response Relationship, Drug; Glutathione; Glutathione Peroxidase; Glutathione Reductase; Homovanillic Acid; Male; Motor Activity; Neuroprotective Agents; Oxidopamine; Parkinson Disease, Secondary; Rats; Rats, Wistar; Substantia Nigra; Superoxide Dismutase; Thiobarbituric Acid Reactive Substances; Vitamin A

The main objective of the study was to examine whether crocetin, a natural product from Gardenia jaminoides Ellis, has beneficial effects on the state of insulin resistance induced by dexamethasone in a rat model. Measured using the oral glucose tolerance tests (OGTT), male Wistar rats treated with subcutaneous dexamethasone (0.08 mg/kg/d) for 6 weeks exhibited reduced insulin sensitivity at weeks 2 and 4 and impaired glucose tolerance at week 4. In the dexamethasone-treated group, serum insulin, free fatty acids (FFA), triglyceride (TG) and tumor necrosis factor (TNF)-alpha levels were significantly increased at the end of the study. In addition, the hepatic glycogen content was reduced as indicated by periodic acid-Schiff (PAS) staining, and pancreatic islet beta cells showed compensatory hyperactivity suggested by immunohistochemical (IHC) staining using an antibody against insulin. Treatment with crocetin (40 mg/kg/d) significantly attenuated all the described effects of dexamethasone. These results suggest that crocetin might prevent the development of dexamethasone-induced insulin resistance and related abnormalities in rats.

Topics: Administration, Oral; Animals; Blood Glucose; Carotenoids; Dexamethasone; Disease Models, Animal; Fruit; Gardenia; Glucose Tolerance Test; Hypoglycemic Agents; Insulin Resistance; Male; Phytotherapy; Plant Extracts; Rats; Rats, Wistar; Vitamin A

To evaluate the protection afforded by trans-sodium crocetinate against dysoxia in an animal model of recurrent sub-diaphragmatic ischaemia.. Prospective experimental animal study.. University research laboratory. Adult male Sprague-Dawley rats.. Twelve adult male Sprague-Dawley rats (340-510 g) were anaesthetised with sodium pentobarbitone 60 mg/kg i.p. and ventilated with oxygen and isoflurane via tracheostomy. Six 2-min episodes of sub-diaphragmatic hypotension (mean pressure 30 mmHg) were induced using a sling around the proximal aorta. Before the third and sixth episodes, saline 1.5 ml/kg was injected into the aortic cannula. In six rats, this saline contained trans-sodium crocetinate 50 microg/ml.. Ileal luminal PCO(2) and distal aortic pressure were monitored continuously. Following ischaemic episodes trans-sodium crocetinate had no discernible effect on either degree of PCO(2) elevation or the time to peak PCO(2). No effects on renal energy charge or nucleotide concentrations were detected. UV-visible spectroscopy of the crocetinate preparation showed that some cis isomer was present.. The findings, although limited to one drug dosage in one animal model, bring into question whether trans-sodium crocetinate affects plasma oxygen diffusivity in vivo. Alternative explanations for the negative findings include a TSC-induced exacerbation of arterio-venous oxygen shunting, the brevity of the dysoxic episodes, and the presence of cis isomer.

Topics: Animals; Carbon Dioxide; Carotenoids; Disease Models, Animal; Hypercapnia; Intestinal Mucosa; Ischemia; Kidney; Male; Prospective Studies; Rats; Rats, Sprague-Dawley; Vitamin A

This is a review of studies with two agents, glutamine and crocetin, which have been found to enhance recovery of cellular adenosine triphosphate (ATP) and adenosine diphosphate after hemorrhagic shock.. The studies used a sublethal (30 minutes) reservoir shock model in 300- to 350-g, male, Sprague-Dawley rats, using either ketamine-xylazine or isoflurane anesthesia. Glutamine was given as a 3% (21 mmol/L) solution in Ringer's lactate (630 mg/kg). Crocetin was given as a 500 nmol/L solution in Ringer's lactate (2 mg/kg).. Both glutamine and crocetin caused recovery of ATP to baseline levels (9.0 micromol/g) within 60 to 120 minutes after resuscitation. Xanthine levels returned more rapidly to baseline (0.1 micromol/g). Both agents prevented the elevation in apoptosis seen in controls at 24 and 48 hours.. Glutamine is a metabolic substrate and a precursor of ATP synthesis. Crocetin enhances oxygen diffusivity in plasma. Both agents restore cellular energy stores to normal after hemorrhagic shock and produce a marked diminution in the extent of apoptosis postshock. Their mechanism of action probably involves prevention of mitochondrial damage.

Topics: Adenosine Diphosphate; Adenosine Triphosphatases; Animals; Antioxidants; Apoptosis; Carotenoids; Disease Models, Animal; Energy Metabolism; Fluid Therapy; Glutamine; Intestine, Small; Liver; Male; Rats; Rats, Sprague-Dawley; Resuscitation; Shock, Hemorrhagic; Vitamin A

The purpose of this study was to evaluate the effects of extracts of Coptidis rhizoma, Phellodendri cortex and Gardeniae fructus, which are medicinal herbs in Orengedoku-to (Huanglin-Jie-Du-Tang in Chinese), and crocetin (a major component of Gardeniae fructus) on experimental elevation of aqueous flare in pigmented rabbits. To produce aqueous flare elevation, 0.5 microg/kg lipopolysaccharide (LPS) was injected into the ear vein, or prostaglandin E2 (PGE2) 25 microg/ml, was applied to the cornea by means of a glass cylinder. Animals were pretreated by oral administration of 150 g/day of food containing 0.15% (w/w) extract powder of Coptidis rhizoma, 0.10% (w/w) extract powder of Phellodendri cortex or 0.15% (w/w) extract powder of Gardeniae fructus for 4 days, or by intravenous injection of crocetin, 0.3, 3, 30 or 300 microg/kg, 30 minutes before aqueous flare elevation. Aqueous flare was measured with a laser flare-cell meter. Aqueous flare intensity was expressed as the area under the curve (AUC) in arbitrary units. The AUC of LPS- and PGE2-induced aqueous flare elevation was 4685 and 1386 arbitrary units, respectively. Pretreatment by oral administration of 0.15% (w/w) extract of Coptidis rhizoma or 0.10% (w/w) extract of Phellodendri cortex did not inhibit LPS-induced aqueous flare elevation. Pretreatment by oral administration of 0.15% extract of Gardeniae fructus suppressed LPS-induced aqueous flare elevation (AUC: 1411 arbitrary units). Pretreatment by intravenous injection of 3, 30 or 300 microg/kg of crocetin-inhibited LPS-induced aqueous flare elevation in a dose-dependent manner. Pretreatment with 3 or 30 microg/kg of crocetin did not inhibit PGE2-induced aqueous flare elevation, but 300 microg/kg of crocetin inhibited PGE2-induced aqueous flare elevation (AUC: 918 arbitrary units).

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Aqueous Humor; Area Under Curve; Carotenoids; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Injections, Intravenous; Lipopolysaccharides; Male; Rabbits; Time Factors; Uveitis, Anterior; Vitamin A