Biochanin A attenuates myocardial ischemia/reperfusion injury through the TLR4/NF-κB/NLRP3 signaling pathway

Abstract Purpose: Myocardial ischemia/reperfusion (Ml/R) injury is a leading cause of damage in cardiac tissues, with high rates of mortality and disability. Biochanin A (BCA) is a main constituent of Trifolium pratense L. This study was intended to explore the effect of BCA on Ml/R injury and explore the potential mechanism. Methods: In vivo MI/R injury was established by transient coronary ligation in Sprague-Dawley rats. Triphenyltetrazolium chloride staining (TTC) was used to measure myocardial infarct size. ELISA assay was employed to evaluate the levels of myocardial enzyme and inflammatory cytokines. Western blot assay was conducted to detect related protein levels in myocardial tissues. Results: BCA significantly ameliorated myocardial infarction area, reduced the release of myocardial enzyme levels including aspartate transaminase (AST), creatine kinase (CK-MB) and lactic dehydrogenase (LDH). It also decreased the production of inflammatory cytokines (IL-1β, IL-18, IL-6 and TNF-α) in serum of Ml/R rats. Further mechanism studies demonstrated that BCA inhibited inflammatory reaction through blocking TLR4/NF-kB/NLRP3 signaling pathway. Conclusion: The present study is the first evidence demonstrating that BCA attenuated Ml/R injury through suppressing TLR4/NF-kB/NLRP3 signaling pathway-mediated anti-inflammation pathway.

Research on Etherification Reaction of Three Kinds of Isoflavones from Chickpeas / 世界科学技术-中医药现代化

This study was aimed to improve the drug activity of three kinds of isoflavones from Chickpeas.Biochanin A,formononetin and genistein were used as raw materials.Acetone was used as solvent.Potassium carbonate was used as catalyst.The etherification reaction was with 1,3-dibromopropane,1-bromopropane and 3-bromopropene.The results showed that 9 isoflavone ramifications were synthesized.This method was simple,easy to control with high yield.It was concluded that the product structure was confirmed by 1H-NMR,13C-NMR and ESI-MS analysis.It laid a foundation for the structural study basis in the further research of its drug activity.

Antioxidant Effects of Biochanin A in Streptozotocin Induced Diabetic Rats

ABSTRACT Bioflavonoid-containing diets have been reported to be beneficial in diabetes. In the current study, the effect of Biochanin A (BCA) on blood glucose, antioxidant enzyme activities and oxidative stress markers in diabetic rats were investigated. 30 male Wistar rats were divided into five groups. Two of them were selected as control; group1: control (receiving 0.5%DMSO), and group2: Control+BCA (receiving 10 mg/kg.bw BCA). Diabetes was induced in other rats with injection of (55 mg/kg.bw) streptozotocin; group3: diabetic control (receiving 0.5%DMSO), groups 4 and 5 were treated with 10 and 15 mg/kg.bw BCA respectively. After 6 weeks the following results were obtained. Fasting blood glucose (FBG), Triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C) and malondialdehyde (MDA) levels significantly increased and body weight, high density lipoprotein cholesterol (HDL-C), superoxide dismutase (SOD) and catalase (CAT) activity and total antioxidant status (TAS) significantly decreased in diabetic rats as compared to control rats. Oral administration of BCA in 10 and 15 mg/kg.bw, FBG, TG, TC, LDL-C, VLDL-C were decreased significantly in all treated rats. MDA was decreased in all treated rats but it was significant just in 15 mg/kg.bw BCA. HDL, CAT, SOD, and TAS were significantly increased in treated group with 15 mg/kg.bw. The obtained results indicated hypoglycemic and hypolipidemic effect of BCA. Also BCA reduced oxidative stress in diabetic rats.

Effect and mechanism of biochanin A on rat hippocampal neurons injuries impaired by H2O2 / 中国药学杂志

OBJECTIVE: To observe the effect of Biochanin A (Bio A) on rat hippocampal neurons injuries impaired by H2O2 and Bcl-2, Bax and caspase-3 protein expression. METHODS: Hippocampal neurons of newly born rat were cultured in vivo and were injured by H2O2. Different concentration of Bio A on cell viability was measured by CCK-8; cell apoptosis rate was tested by flow cytometry analysis; Bcl-2, Bax and caspase-3 protein expression were tested by Western blot method. RESULTS: Compared with the model group, Bio A significantly improved hippocampal neurons cell viability and inhibited cell apoptosis and necrosis. In addition, Bio A clearly enhanced the protein expression of Bcl-2 and decrease the protein expression of Bax and caspase-3. CONCLUSION: The protective effect of Bio A on rat hippocampal neurons injuries impaired by H2O2 may be related to the inhibition of hippocampal neurons apoptosis. The mechanism of Bio A against cell apoptosis may involve the increased Bcl-2 protein expression and reduced Bax, caspase-3 protein expression.

Mechanisms of phytoestrogen biochanin A-induced vasorelaxation in renovascular hypertensive rats

BACKGROUND: The plant-derived estrogen biochanin A is known to cause vasodilation, but its mechanism of action in hypertension remains unclear. This study was undertaken to investigate the effects and mechanisms of biochanin A on the thoracic aorta in two-kidney, one clip (2K1C) renovascular hypertensive rats. METHODS: Hypertension was induced by clipping the left renal artery, and control age-matched rats were sham treated. Thoracic aortae were mounted in tissue baths to measure isometric tension. RESULTS: Biochanin A caused concentration-dependent relaxation in aortic rings from 2K1C hypertensive and sham-treated rats, which was greater in 2K1C rats than in sham rats. Biochanin A-induced relaxation was significantly attenuated by removing the endothelium in aortic rings from 2K1C rats, but not in sham rats. Nomega-Nitro-L-arginine methylester, a nitric oxide synthase inhibitor, or indomethacin, a cyclooxygenase inhibitor, did not affect the biochanin A-induced relaxation in aortic rings from 2K1C and sham rats. By contrast, treatment with glibenclamide, a selective inhibitor of adenosine triphosphate-sensitive K+ channels, ortetraethy-lammonium, an inhibitor of Ca2+-activated K+ channels, significantly reduced biochanin A-induced relaxation in aortic rings from both groups. However, 4-aminopyridine, a selective inhibitor of voltage-dependent K+ channels, inhibited the relaxation induced by biochanin A in 2K1C rats, whereas no significant differences were observed in sham rats. CONCLUSION: These results suggest that the enhanced relaxation caused by biochanin A in aortic rings from hypertensive rats is endothelium dependent. Vascular smooth muscle K+ channels may be involved in biochanin A-induced relaxation in aortae from hypertensive and normotensive rats. In addition, an endothelium-derived activation of voltage-dependent K+ channels contributes, at least in part, to the relaxant effect of biochanin A in renovascular hypertension.

BIOCHANIN A PROTECTS DOPAMINERGIC NEURONS BY INHIBITING MICROGLIA ACTIVATION / 营养学报

Objective:To explore the protective effect of biochanin A(Bioch A) on lipopolysaccharide(LPS) -induced dopaminergic neurodegeneration. Method:An in vitro model of LPS-induced dopaminergic neurodegeneration was used to investigate the protective effect of Bioch A on dopaminergic neurons against LPS-induced neurotoxicity by measuring [3H] DA uptake and counting TH-immunoreactive cells. Microglia were visualized by staining for the CR3 complement receptor with monoclonal antibody OX-42. The production of nitric oxide(NO) was determined by measuring the accumulated levels of nitrite in the culture supernatant with the Griess reagent,and release of tumor necrosis factor-?(TNF-?) was measured by immunosorbent assay. The production of superoxide was determined by measuring the superoxide dismutase(SOD) -inhibitable reduction of cytochrome C. Results:BiochA dose-dependently attenuated LPS-induced decrease in dopamine(DA) uptake and the number of dopaminergic neurons in rat mesencephalic neuron-glia cultures. BiochA also significantly inhibited LPS-induced activation of microglia and production of TNF-?,NO and superoxide in rat mesencephalic neuron-glia cultures and microglia-enriched cultures. Conclusion:Biochanin A may protect dopaminergic neurons from LPS-induced injury and its effectiveness in inhibiting microglia activation may underlie one of the mechanisms.

Inhibitory effects of biochanin A on mouse lung tumor induced by benzo(a)pyrene

Biochanin A, an isoflavone compound, is reported to have an inhibitory effect on benzo(a)pyrene [B(a)P] metabolism. We examined the modifying effect of biochanin A on in vivo carcinogenesis using a mouse lung tumor model. As carcinogens, a single subcutaneous injection of 0.5mg of B(a)P was given within 24 hours after birth. The test groups were injected with 0.125mg of biochanin A in 0.1ml DMSO by i.p. 3 times a week for 6 weeks after weaning. All mice were sacrificed at week 9 and the incidence and multiplicity of lung tumors were examined. Concomitant administration of biochanin A showed a significant inhibitory effect on the incidence of tumor-bearing mice (12.5%, P<0.01), as well as the mean number of tumors (0.13, P<0.001), compared with the group treated with B(a)P alone in which the incidence was 57.1% and the mean number was 1.0. These results suggest that biochanin A has inhibitory potential on the development of mouse lung tumor induced by B(a)P.