Extration Of Black Bean Hull Derived Active Constituents And Its Effect On Kidney Injury Repair

Black soybean hull are one of the traditional medicine and food homogenous agricultural resources.At present,the research on black beans mainly focuses on the study of proteins,and there are few reports on other biological activities.In this paper,black soybean hull was used as raw material to study the extraction process of anthocyanins from black soybean hull.The anthocyanins of black soybean hull were isolated and purified by macroporous adsorption resin.The composition of anthocyanins in black soybean hull was analyzed by UV,IR,HPLC,and paper chromatography.The study of black soybean hull anthocyanin on the repair of rat kidney injury was studied.This provides a certain reference for the further development and utilization of black bean skin.Comparing five methods to extract anthocyanins,it was found that the extraction rate of anthocyanins in black bean hulls was the highest by ethanol / ammonium sulfate aqueous two phase system.The optimum conditions for the extraction of aqueous two phase were determined by single factor test.The response surface methodology was used to optimize the extraction of anthocyanins from black soybean hull by aqueous two phase.The optimal parameters were optimized by response surface analysis.The liquid ratio,ammonium sulfate massfraction,ethanol concentration,p H were 1:56(g/m L),22 %,30 %,3.0 respectively.The optimal yield of anthocyanin was 2.81 mg/g.Two kinds of macroporous adsorption resins were used for the static adsorption of the anthocyanin extract from Black Soybean Hull,and AB-8 type resin presented the highest adsorption ratio and was consequently selected for further investigations.The optimum conditions for the dynamic adsorption and desorption of the anthocyanin extract were explored.Lyophilization gave a crystallized anthocyanin.The major components of the anthocyanin was identified to be ascyanidin-3-O-glucoside and delphinidol-3-O-glucoside through IR spectroscopic,UV-vis,thin layer chromatographic,and HPLC.Study on the effect of anthocyanin from black soybean hulls on cisplatin-induced acute kidney injury in rats by animal experiments.The rats were treated with low(30mg/(kg·d)).medium(150mg/(kg·d)),and high(750mg/(kg·d))three black soybean hull anthocyanins at the same time.The platinum model group gavage distilled water for12 days.On the tenth day of the experiment,the cisplatin model group and the anthocyanin(low,middle,high)group were intraperitioneally injected with cisplatin7.5mg/Kg(by body weight),and normal rats were injected with the same amount of normal saline..Determination of serum urea nitrogen(BUN)and creatinine(Scr)levels?pathology and Western bolt in each group.Results:Compared with the normal groupthe,anthocyanin(low,middle,high)group of black soybean hull significantly inhibited the increase of serum urea nitrogen(BUN)and creatinine(Scr)in rats induced by cisplatin;HE staining revealed that renal tubular injury was severe,medullary hyperemia,renal tubular structure was destroyed,epithelial cells were detached,and vacuolar degeneration was observed in the model group.The kidneys of the black soybean anthocyanin group were alleviated.Western blot showed that the expression of Bcl-2,Bax,Caspase-3 was up regulated respectively at Model group,while the anthocyanin(low,middle,high)group could prevent the changes.Conclusion:Anthocyanin from black soybean hull has a protective effect on kidney injury induced by cisplatin in rats.The mechanism may be related to the decrease of lipid peroxides and increase of antioxidant enzyme activity in black soybean hull anthocyanins.