aurapten and Disease-Models--Animal

In the 1980s, the authors developed the enzyme immunoassay (EIA) system for mouse nerve growth factor (NGF) to clarify its important physiological roles. Our EIA system was a new and powerful tool for measurement of extremely low levels of NGF in vitro and in vivo, and it contributed to investigation into the regulatory mechanism of NGF synthesis. After that, we demonstrated that the compounds with a low molecular weight, such as 4-methylcatechol, which elicit stimulatory activity toward NGF synthesis, were useful and practical for therapeutic purposes; as NGF has potent activity on neuronal degeneration in both the central nervous system (CNS) and the peripheral nervous system. Since 2008, we have been searching for and isolating neuroprotective component(s) from citrus peels. As a result, our study revealed that 1) 3,5,6,7,8,3',4'-heptamethoxyflavone (HMF) has neuroprotective ability in the CNS by inducing brain-derived neurotrophic factor (BDNF) and by suppressing inflammation; 2) auraptene (AUR) also has neuroprotective ability in the CNS by suppressing inflammation and by probably inducing neurotrophic factor(s). As the content of AUR in the peels of Kawachi Bankan is exceptionally high, 1) we found this peel powder to exert neuroprotective effects in the brain of various pathological model mice; 2) some of the AUR transited from the peel to the juice during the squeezing process to obtain the juice. Therefore, K. Bankan juice, which is enriched in AUR by adding peel paste to the raw juice, was shown to be practical for suppression of cognitive dysfunction of aged healthy volunteers.

Topics: Animals; Catechols; Citrus; Cognitive Dysfunction; Coumarins; Disease Models, Animal; Drug Discovery; Female; Humans; Immunoenzyme Techniques; Male; Mice; Nerve Growth Factor; Neuroprotective Agents; Phytotherapy; Rats

We have recently shown that 3,5,6,7,8,3',4'-heptamethoxyflavone (HMF) and auraptene (AUR) have neuroprotective effects on the central nervous system. HMF, a citrus flavonoid, altered NMDA-type glutamate receptor antagonist MK-801-induced memory dysfunction and schizophrenia-positive symptom-like behavior. HMF also showed a protective effect against ischemia-induced short-term memory dysfunction. In the ischemic brain, HMF induced the following protective effects against brain dysfunction: 1) rescue of neuronal cell death in the hippocampus; 2) increased production of brain-derived neurotrophic factor; 3) stimulation of neurogenesis in the dentate gyrus subgranular zone; 4) activation of the autophosphorylation of calcium-calmodulin-dependent protein kinase II; and 5) suppression of microglial activation. On the other hand, AUR, a citrus coumarin, ameliorated lipopolysaccharide-induced inflammation in the brain as shown by inhibition of microglial activation and inhibition of cyclooxygenase (COX)-2 expression in the hippocampus. AUR also showed antiinflammatory effects on the ischemic brain by inhibiting microglial activation, COX-2 expression, and neuronal cell death in the hippocampus. The peel of kawachibankan (Citrus kawachiensis), a noted citrus product of Ehime prefecture, Japan, contains AUR, HMF, naringin, and narirutin. The dried powder of both the peel and juice had antiinflammatory effects in the mouse hippocampus, suggesting that citrus compounds may be beneficial as neuroprotective agents in the treatment of neurological disorders.

Topics: Animals; Anti-Inflammatory Agents; Brain; Brain Ischemia; Brain-Derived Neurotrophic Factor; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cell Death; Central Nervous System; Citrus; Coumarins; Cyclooxygenase 2; Disease Models, Animal; Flavones; Food; Hippocampus; Humans; Memory Disorders; Mice; Microglia; Nervous System Diseases; Neurogenesis; Neuroprotective Agents; Phytotherapy; Receptors, N-Methyl-D-Aspartate; Schizophrenia

The main strategy of cancer cells for survival is uncontrolled cell division and escape from apoptosis. The use of anticancer agents inducing the production of reactive oxygen species (ROS) and controlling cell division might be a therapeutic approach to eradicate cancer cells. Herein, we examined the therapeutic effects of Auraptene on CT26 cells as well as on a mouse model of colorectal cancer (CRC). The spheroid assay was also conducted to analyze the anti-proliferative activity of Auraptene. We also assessed the in vitro analysis of ROS generation. The impact of Auraptene on oxidant/antioxidant markers, as well as the mRNA expression of Bax, Bcl-2, Nrf2, Cyclin D1, and Survivin genes, was evaluated by qPCR in tumor samples. As a result, Auraptene significantly reduced the size of CT26 spheroids at a dose of 200 µM. After 12 h, ROS levels were significantly elevated in CT26 cells. The administration of Auraptene induced apoptosis and the cell cycle arrest by modulating Bax, Bcl-2, Nrf2, Cyclin D1, and Survivin mRNA levels. Furthermore, our results demonstrated that Auraptene suppressed CAT, GSH (reduced Glutathione), and FRAP while increasing MDA in tissue homogenates which in turn could raise oxidative stress and stimulate apoptosis. Therefore, Auraptene may act as a powerful adjuvant therapy in CRC since it triggers apoptosis and cell cycle.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Cell Proliferation; Colorectal Neoplasms; Coumarins; Cyclin D1; Disease Models, Animal; Mice; NF-E2-Related Factor 2; Oxidative Stress; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Survivin

Farnesoid X receptor (FXR) has been considered as a promising target for nonalcoholic steatohepatitis (NASH), while existing FXR agonists suffer from serious side effects. Thus, it is very necessary to identify novel FXR agonists with good safety. Auraptene (AUR) is a new FXR agonist with excellent safety and extensive pharmacological activities, while the lactone of AUR is vulnerable to esterolysis. In this study, the lactone of AUR was converted to metabolically stable amide moiety, and the obtained analog SU5 revealed comparable activity and better metabolic stability than that of AUR. In NASH model, SU5 showed stronger efficacy than AUR on fatty liver by upregulating gene expressions related to FXR in vivo. Moreover, SU5 improved lipid metabolism by downregulating the gene expressions of lipid synthesis, while upregulating the gene expressions of fatty acid β-oxidation and triglyceride metabolism. Besides, the inflammation-related genes were significantly decreased in SU5-treated group. These positive results highlighted the pharmacological potential of SU5 for the treatment of NASH.

Topics: Animals; Choline; Coumarins; Diet; Disease Models, Animal; Down-Regulation; Half-Life; Humans; Lipid Metabolism; Liver; Methionine; Mice; Mice, Obese; Non-alcoholic Fatty Liver Disease; Rats; Receptors, Cytoplasmic and Nuclear; Tumor Necrosis Factor-alpha; Up-Regulation

Benign prostatic hyperplasia (BPH) is a disease that commonly strikes the majority of aged men. Developing new therapies to manage BPH with improved efficacy and safety is strongly needed. In this regard, auraptene is a natural compound with multiple pharmacological effects, but with poor oral bioavailability. This investigation aimed to assess the possible protection offered by auraptene-nanostructured lipid carrier (auraptene-NLC) in a BPH model induced by testosterone in rats. Auraptene-NLC had optimum particle size and drug release profile compared to raw auraptene. At doses (5 and 10 mg/kg), it hampered the rise in prostatic weights & indices relative to rats challenged with testosterone. Moreover, auraptene-NLC alleviated histopathological abnormalities in prostate architecture and decreased the glandular epithelial height. Additionally, testosterone-induced oxidative stress was alleviated by auraptene-NLC and inhibited raised lipid peroxidation, catalase and superoxide dismutase exhaustion as well as enhanced glutathione content. Moreover, it significantly reduced the prostate content of nuclear factor κB, Interleukins1β & 6, as well as transforming growth factor β, compared to testosterone group. The proapoptotic activity of auraptene-NLC (10 mg/kg) was confirmed by a significant increase of prostate cleaved caspase-3, boosted Bax/Bcl2 mRNA ratio that was further confirmed by assessing their protein expressions. Furthermore, the beneficial effects of auraptene-NLC against BPH were substantiated by ameliorating testosterone-induced decline of nuclear PPARα & PPARγ and inhibiting the increased expression of cyclin D1 protein. In conclusion, auraptene-NLC offers a protective effect in rats whereby BPH was induced by testosterone, via its anti-inflammatory, antioxidant and proapoptotic activities, and PPAR family activation.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Apoptosis; Apoptosis Regulatory Proteins; Coumarins; Disease Models, Animal; Drug Compounding; Drug Liberation; Inflammation Mediators; Male; Nanoparticles; Nanotechnology; Oxidative Stress; Peroxisome Proliferator-Activated Receptors; PPAR alpha; PPAR gamma; Prostate; Prostatic Hyperplasia; Rats, Wistar; Signal Transduction; Testosterone

Early life stress is associated with various complications. Auraptene has significant antioxidant and anti-inflammatory effects. This study aimed to assess the probable underlying mechanisms that mediate changes in the behavior, hippocampus, heart and serum in the mouse model of maternal separation (MS) stress. We evaluated the possible protective effects of auraptene in these changes focusing on inflammatory response and oxidative state. Mice were treated with auraptene (5, 10, and 50 mg/kg). In addition, anxiety-like behaviors were evaluated using behavioral tests; including open field test (OFT) and elevated plus maze (EPM). Hippocampus and heart samples were assessed histopathologically. Levels of malondialdehyde (MDA) and antioxidant capacity, as well as nitrite levels, were measured in serum, heart, and hippocampal tissues. Moreover, gene expression of inflammatory markers (Il-1β and Tlr-4) was evaluated in the heart and hippocampus. Results showed that auraptene reversed the negative effects of MS on behavior (increased time spent in central zone of the OFT and time and entries to the open arms of the EPM). Auraptene mitigated adverse effects of MS on the hippocampus (increased diameter and decreased percentage of dark neurons in the CA3 area). Accordingly, auraptene decreased MDA and nitrite levels and increased the antioxidant capacity in serum, and hippocampal samples. However, we observed different effects for different doses of auraptene in the heart samples. We concluded that MS is associated with anxiety-like behavior and cellular/molecular modifications in the heart, hippocampus and serum. We found that auraptene exerted protective effects against these negative effects of MS in mouse.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Anxiety, Separation; Behavior, Animal; Coumarins; Disease Models, Animal; Heart; Hippocampus; Humans; Interleukin-1beta; Maternal Deprivation; Mice; Oxidative Stress; Stress, Psychological

Breast cancer is a common malignant tumor in women with limited treatment options and multiple side effects. Today, the anti-cancer properties of natural compounds have attracted widespread attention from researchers worldwide.. In this study, we treated 4T1 tumor-bearing Balb/c mice with intraperitoneal injection of Auraptene, paraffin oil, and saline as two control groups. Body weight and tumor volume were measured before and after treatment. Hematoxylin and eosin (H & E) staining and immunohistochemistry of Ki-67 were used as markers of proliferation. In addition, ELISA assays were performed to assess serum IFN-γ and IL-4 levels.. There was no significant change in body weight in all animal groups before and after treatment. 10 days after the last treatment, Auraptene showed its anti-cancer effect, which was confirmed by the smaller tumor volume and H & E staining. In addition, Ki-67 expression levels were significantly reduced in tumor samples from the Auraptene-treated group compared to the paraffin oil and saline-treated groups. In addition, in tumor-bearing and normal mice receiving Auraptene treatment, IL-4 serum production levels were reduced, while serum levels of IFN-γ were significantly up-regulated in tumor-bearing mice after Auraptene treatment.. In the case of inhibition of tumor volume and Ki-67 proliferation markers, Auraptene can effectively inhibit tumor growth in breast cancer animal models. In addition, it might increases Th1 and CD8 + T cell responses after reducing IL-4 serum levels and IFN-γ upregulation, respectively. However, further research is needed to clarify its mechanism of action.

Topics: Allografts; Animals; Antineoplastic Agents; Biological Products; Biomarkers, Tumor; Breast Neoplasms; Cell Line, Tumor; Coumarins; Cytokines; Disease Models, Animal; Female; Immunohistochemistry; Mice; Mice, Inbred BALB C; Molecular Structure; Tumor Burden

Polycystic ovary with poor-quality oocytes has remained problematic in polycystic ovary syndrome (PCOS) patients. It is well documented that the inflammation and production of reactive oxygen species (ROS) in PCOS ovaries are significantly higher than normal voluntaries. In this study, we hypothesized that auraptene (AUR), as a coumarin derivative with anti-inflammatory properties, may be effective in improvement of oocyte maturation and fertilization rate in PCOS patients. For this purpose, PCOS model was induced in NMRI mice and confirmed by ovarian histopathology observations and hormonal assays. PCOS-induced mice were administrated with AUR (PCOS-AUR) and metformin (PCOS-MET), and their effects on inflammation, apoptosis rate, oocyte maturation, and in vitro fertilization capacity were determined and compared with those normal and PCOS animals treated with sesame oil (PCOS-sesame oil) and no treatment (PCOS). Treatment with AUR and MET decreased the inflammation and apoptosis rates in PCOS mice compared with PCOS animals with no treatment. PCOS-AUR and PCOS-MET oocytes also showed higher intracellular glutathione and lower ROS concentrations compared with PCOS mice, indicating improved oocyte maturation rate. PCOS-AUR and PCOS-MET groups showed higher percentages of expansion rate and MII stage oocytes, and lower rate of abnormal oocytes compared with PCOS with no treatment. The rate of fertilization in the oocytes isolated from PCOS-AUR and PCOS-MET groups was higher than PCOS-sesame oil and PCOS groups. Our findings suggest that AUR can be considered as a potential candidate for improvement of oocyte maturation and fertilization capacity in PCOS patients, comparable to MET.

Topics: Animals; Coumarins; Cumulus Cells; Disease Models, Animal; Estradiol; Female; Fertilization; Fertilization in Vitro; Inflammation; Mice; Oocytes; Oogenesis; Ovary; Polycystic Ovary Syndrome; Progesterone; Reactive Oxygen Species; Testosterone; Tumor Necrosis Factor-alpha

Topics: Animals; Anti-Inflammatory Agents; Brain; Cell Line, Tumor; Cell Survival; Citrus; Coumarins; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation, Neoplastic; Glial Cell Line-Derived Neurotrophic Factor; Glioma; Mice; Nerve Growth Factors; Neuroprotective Agents; Phosphorylation; Rats; RNA, Messenger; Signal Transduction

Current therapeutics for Parkinson's disease (PD) are only effective in providing relief of symptoms such as rigidity, tremors and bradykinesia, and do not exert disease-modifying effects by directly modulating mitochondrial function. Here, we investigated auraptene (AUR) as a potent therapeutic reagent that specifically protects neurotoxin-induced reduction of mitochondrial respiration and inhibits reactive oxygen species (ROS) generation. Further, we explored the mechanism and potency of AUR in protecting dopaminergic neurons. Treatment with AUR significantly increased the viability of substantia nigra (SN)-derived SN4741 embryonic dopaminergic neuronal cells and reduced rotenone-induced mitochondrial ROS production. By inducing antioxidant enzymes AUR treatment also increased oxygen consumption rate. These results indicate that AUR exerts a protective effect against rotenone-induced mitochondrial oxidative damage. We further assessed AUR effects in vivo, investigating tyrosine hydroxylase (TH) expression in the striatum and substantia nigra of MPTP-induced PD model mice and behavioral changes after injection of AUR. AUR treatment improved movement, consistent with the observed increase in the number of dopaminergic neurons in the substantia nigra. These results demonstrate that AUR targets dual pathogenic mechanisms, enhancing mitochondrial respiration and attenuating ROS production, suggesting that the preventative potential of this natural compound could lead to improvement in PD-related neurobiological changes.

Topics: Animals; Behavior, Animal; Biomarkers; Cell Respiration; Coumarins; Disease Models, Animal; Dopaminergic Neurons; Free Radical Scavengers; Gene Expression; Mice; Mitochondria; Models, Biological; Oxidation-Reduction; Parkinson Disease; Reactive Oxygen Species; Tyrosine 3-Monooxygenase

We investigated the effects of auraptene on mouse oligodendroglial cell lineage in an animal model of demyelination induced by cuprizone. Auraptene, a citrus coumarin, was intraperitoneally administered to mice fed the demyelinating agent cuprizone. Immunohistochemical analysis of the corpus callosum and/or Western blotting analysis of brain extracts revealed that cuprizone reduced immunoreactivity for myelin-basic protein, a marker of myelin, whereas it increased immunoreactivity to platelet derived-growth factor receptor-α, a marker of oligodendrocyte precursor cells. Administration of auraptene enhanced the immunoreactivity to oligodendrocyte transcription factor 2, a marker of oligodendrocyte precursor cells and oligodendrocyte lineage precursor cells, but had no effect on immunoreactivity to myelin-basic protein or platelet-derived growth factor receptor-α. These findings suggest that auraptene promotes the production of oligodendrocyte lineage precursor cells in an animal model of demyelination and may be useful for individuals with demyelinating diseases.

Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Brain; Coumarins; Cuprizone; Demyelinating Diseases; Disease Models, Animal; Drug Evaluation, Preclinical; Gene Expression; Injections, Intraperitoneal; Male; Mice, 129 Strain; Mice, Inbred C57BL; Microglia; Myelin Basic Protein; Nerve Tissue Proteins; Neuroprotective Agents; Oligodendrocyte Transcription Factor 2; Oligodendroglia; Receptor, Platelet-Derived Growth Factor alpha; Stem Cells

The anti-inflammatory activity of auraptene (AUR), a citrus coumarin, in peripheral tissues is well-known, and we previously demonstrated that AUR exerts anti-inflammatory effects in the ischemic brain; the treatment of mice with AUR for eight days immediately after ischemic surgery suppressed demise and neuronal cell death in the hippocampus, possibly through its anti-inflammatory effects in the brain. We suggested that these effects were at least partly mediated by the suppression of inflammatory mediators derived from astrocytes. The present study showed that (1) AUR, as a pretreatment for five days before and another three days after ischemic surgery, suppressed microglial activation, cyclooxygenase (COX)-2 expression in astrocytes, and COX-2 mRNA expression in the hippocampus; (2) AUR suppressed the lipopolysaccharide-induced expression of COX-2 mRNA and the mRNA of pro-inflammatory cytokines in cultured astrocytes; (3) AUR was still detectable in the brain 60 min after its intraperitoneal administration. These results support our previous suggestion that AUR directly exerts anti-inflammatory effects on the brain.

Topics: Animals; Anti-Inflammatory Agents; Brain; Brain Ischemia; Coumarins; Cyclooxygenase 2; Cytokines; Disease Models, Animal; Gene Expression Regulation; Glial Fibrillary Acidic Protein; Hippocampus; Inflammation Mediators; Male; Mice

Cerebral ischemia causes delayed neuronal cell death in the hippocampus resulting in sequential cognitive impairments. Hyper-activated inflammation following ischemia is one of the etiologies for delayed neuronal cell death. In the present study, using a transient global ischemia mouse model, we showed that auraptene (AUR), a citrus coumarin, effectively inhibited microglia activation, cyclooxygenase-2 expression by astrocytes, and neuronal cell death in the hippocampus following ischemic insults. These results suggest that AUR acts as a neuroprotective agent in the ischemic brain, which may be mediated by suppression of the inflammatory response.

Topics: Animals; Anti-Inflammatory Agents; Astrocytes; Brain Ischemia; Cell Death; Coumarins; Cyclooxygenase 2; Disease Models, Animal; Hippocampus; Inflammation; Male; Mice; Mice, Inbred C57BL; Microglia; Neurons; Neuroprotective Agents