Mechanism Study on Apoptosis of Osteoclasts Induced by Guilu Erxian Jiao(龟鹿二仙胶)
Objective To investigate the effects of different concentration of Guilu Erxian Jiao(龟鹿二仙胶)on osteoclast ap-optosis and osteoclast relevant mechanism.Methods RAW264.7 cells were cultured and passaged and RAW264 in mouse mono-cytes was induced using M-CSF+RANKL 7 cell lines to differentiate into osteoclasts and obtain osteoclasts.CCK8 method,TUNEL staining and low cytometric analysis were used to detect the effects of different concentrations of Guilu Erxian Jiao on os-teoclast apoptosis.The expressions of apoptosis-related molecules Bcl-2,Bax and Cytochrome C were detected by Real-time PCR and Western blot.After the intervention of Guilu Erxian Jiao and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway inhibitor LY294002,the apoptosis rate of osteoclasts and apoptosis-related protein AKT were detected by flow cytometry and Western blot.Results Different concentrations of Guilu Erxian Jiao(high,medium,low dose)could inhibit prolifer-ative activity of osteoclasts,improve the apoptosis rate,the ratio of Bax/Bcl-2 and Cytochrome C expression.But the effect of Guilu Erxian Jiao at medium concentration for 72 h was the most significant.After the intervention of medium concentration of Guilu Erxian Jiao and PI3K/AKT pathway inhibitor LY294002,the phosphorylation of PI3K/AKT pathway related protein AKT protein was significantly inhibited,and the total apoptosis rate of osteoclasts was significantly increased.Conclusion Guilu Erxian Jiao can promote apoptosis of osteoclasts by inhibiting PI3K/AKT pathway activation.
traditional Chinese medicineGuilu Erxian Jiao(龟鹿二仙胶)PI3K/AKTosteoclastsapoptosis
万方数据
维普
