基于PI3K/Akt信号通路介导的细胞凋亡探讨健步汤抗骨质疏松症的作用OA

AIM To investigate the anti-osteoporosis(OP)effects of Jianbu Decoction.METHODS Eighteen rats were selected to establish OP models by bilateral ovariectomy.After successful modeling,they were randomly divided into model group,Xianling Gubao group(0.27 g/kg)and Jianbu Decoction group(15.75 g/kg),with 6 rats in each group.The blank group and the sham operation group were also set up,with 6 rats in each group.After continuous administration for 90 days,Micro-CT was used to detect changes in bone microstructure.ELISA was employed to measure the serum levels of bone formation marker PINP,bone resorption marker CTX-1,and inflammatory factors TNF-α and IL-6 in rats.Flow cytometry was used to detect the apoptosis rate of femoral tissue cells.RT-qPCR was applied to determine the mRNA expressions of PI3K,Akt and PTEN in rat femoral tissue.Dexamethasone-induced MC3T3-E1 cells were used to establish the in vitro OP model,which was divided into blank group,model group,and low,medium and high concentration groups of Jianbu Decoction-containing serum(Jianbu Decoction-L,M,H groups)(10%,15%,20%drug-containing serum).After intervention,CCK-8 assay was used to detect cell proliferation activity.Flow cytometry was used to measure cell apoptosis rate.Western blot was performed to detect the protein expressions of PI3K,p-PI3K,Akt,p-Akt,Bax and Bcl-2.RT-qPCR was used to determine the mRNA expressions of PI3K,Akt,BGP,Runx2 and COL1a1in cells.RESULTS Jianbu Decoction could increase bone mineral density,bone volume fraction and trabecular number in OP rats(P＜0.05),elevate PINP level(P＜0.05),reduce the levels ofCTX-1,TNF-α and IL-6(P＜0.05),decrease the apoptosis rate of cells in femoral tissue(P＜0.05),downregulate PTEN mRNA expression(P＜0.05),and upregulate PI3K and Akt mRNA expressions(P＜0.05).Jianbu Decoction-containing serum could enhance the proliferation activity of MC3T3-E1 cells(P＜0.05),reduce cell apoptosis rate(P＜0.05),increase the protein expressions of p-PI3K,p-Akt and Bcl-2(P＜0.05),decrease Bax protein expression(P＜0.05),and upregulate the mRNA expressions of PI3K,Akt,BGP,Runx2 and COL1a1(P＜0.05).CONCLUSION Jianbu Decoction may exert an anti-osteoporosis effect by activating the PI3K/Akt signaling pathway,promoting the proliferation and activity of MC3T3-E1 cells,reducing cell apoptosis,increasing bone mineral density and alleviating the body's inflammatory response.