Study on mechanism of action of biochanin A on osteoarthritis in vitro
Objective To investigate the mechanism of action of biochanin A(BCA)on osteoarthritis through in vitro experiments.Methods Primary rat chondrocytes were extracted and cultured,and then divided into the control group(without inducer or interferant),the model group[10 ng/mL interleukin-1 β(IL-1 β)induction],and the BCA group(10 ng/mL IL-1β induction+0,3,6,12,24,48 μmol/L BCA intervention).Cell activity was detected by methyl thiazolyl tetrazolium(MTT)method.Senescence-associated β-galactosidase(SA-β-gal)staining was used to observe cell senescence.Intracellular inflammatory factors and oxidative stress indexes were detected by enzyme-linked immunosorbent assay(ELISA),the levels of related proteins were detected by Western blot,and the affinity between BCA and Nrf2 was verified by molecular docking technology.Results After screening the concentration of BCA with MTT,6,12 and 24 μmol/L BCA were selected for subsequent experiments.Compared with the control group,the chondrocyte activity and the activities of catalase(CAT)and superoxide dismutase(SOD)in the model group were significantly decreased(P＜0.05).The number of SA-β-gal positive cells and expres-sion levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and malondialdehyde(MDA)increased significantly(P＜0.05).The expression levels of type Ⅱ collagen(COL2A1)and aggrecan(ACAN)were significantly decreased(P＜0.05).Compared with the model group,the above conditions were significantly reversed in the BCA group(P＜0.05).In addition,BCA could significantly reduce the degrada-tion of inhibitor kappa B alpha(IκB-α)and nuclear translocation of NF-KBp65,and promote the expression of Nrf2 in the nucleus and the expression of human heme oxygenase-1(HO-1)in whole cells(P＜0.05).The results of molecular docking further confirmed that BCA had the highest affinity with Nrf2.Conclusion BCA can inhibit oxidative stress and inflammation of osteoarthritis chondrocytes by regulating Nrf2/NF-KB signaling pathway in vitro,and alleviate joint injury.
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维普
