Effect of Bushen Huoxue(补肾活血)recipe on autophagy of ovariectomized rat chondrocytes based on Akt/mTOR signaling pathway
Objective To investigate whether Bushen Huoxue recipe can protect articular cartilage by regulating Akt/mTOR signaling pathway to promote the autophagy of chondrocytes in ovariectomized rats.Methods Among 30 SPF 12-week-old female SD rats weighing(247.0±7.0)g,6 were randomly selected as the blank control group,and the remaining rats were randomly divided into model group,BSHXR-L group,BSHXR-M group and BSHXR-H group,with 6 rats in each group.The protective effect of Bushen Huoxue recipe on articular cartilage injury in rats was determined by visual observation score,mus-covine O-solid green staining and immunohistochemistry.The expression of autophagy related proteins was detected by West-em-blot,and the relative expression of Akt,mTOR and downstream autophagy genes was detected by qPCR.Results After modeling,BSHXR(L,M,H)groups could alleviate the histological damage of cartilage.Immunohistochemistry showed that the expression of Collagen-Ⅱ and Aggrecan gradually increased,and the expression of MMP-13 gradually decreased,and the differences between BSHXR-M and BSHXR-H groups and model group were statistically significant(P＜0.05).The results of Western-blot showed that the autophagy pathway proteins p-Akt/Akt and p-mTOR/mTOR were inhibited in the BSHXR(L,M,H)groups,and the expressions of downstream proteins Beclin-1 and LC3 Ⅱ were gradually increased,while p62 was gradu-ally decreased,showing a dose effect.QPCR results showed that BSHXR(L,M,H)groups could promote the relative expres-sion of Beclin-1 and LC3 Ⅱ mRNA,and inhibit the relative expression of p62,Akt,mTOR mRNA,and the differences were statistically significant compared with model group(P＜0.05).Conclusion Bushen Huoxue recipe can enhance the cartilage autophagy response by inhibiting the Akt/mTOR signaling pathway,and then protect the cartilage.
万方数据
维普
