Mechanism Of Siwu Decoction Regulating Proliferation And Apoptosis Of The Bone Marrow Stromal Cells Based On PI3K-Akt-FoxO Signal Pathway

Objective:To observe the effect of Siwu decoction on the proliferation and apoptosis of OP9 cells in bone marrow stromal cells of mice.To detect the influence of Siwu decoction on the gene expressions of PI3K,Akt,FoxO,CyclinD1,p27^KIP1 and Bim and the phosphorylation level of Akt and FoxO protein in OP9 cells.Then,to observe the changes of these indices by treated with PI3K inhibitor.To explore the mechanism of Siwu decoction in regulating the proliferation and apoptosis of OP9 cells.Methods:The effects of Siwu decoction with different concentrations on the proliferation of OP9 were detected by MTT,and the effects of Siwu decoction on cell cycle and apoptosis of OP9 cells were observed by flow cytometry.The expression of PI3K,Akt,FoxO,CyclinD1,p27^KIP1 mRNA was examined by real-time fluorescent quantitative PCR,the protein expression of PI3K、Akt、FoxO,CyclinD1,p27^KIP1,Bim and the phosphorylation level of Akt and FoxO were examined by western blot,and the protein expression of PI3K,Akt,CyclinD1,p27^KIP1,Bim and phosphorylation level of Akt and FoxO were examined by immunohistochemistry.Then,by treated with PI3K inhibitor LY294002,the changes of indices above were observed to verify the effect of Siwu decoction on PI3K-Akt-FoxO signal pathway.Results:（1）Siwu decoction promoted the proliferation of OP9 cells.（2）Siwu decoction promoted OP9 cells from G₀/G₁ phase to s phase,and inhibited apoptosis significantly.（3）Siwu decoction upregulated the gene expression of PI3K,Akt and CyclinD1,downregulated the gene expression of FoxO,p27^KIP1,Bim significantly,and raised the phosphorylation level of Akt and FoxO in OP9cells.（4）Siwu decoction upregulated significantly the protein expression of PI3K,Akt,p-Akt,p-FoxO and CyclinD1 and downregulated significantly the protein expression of p27^KIP1 and Bim.（5）LY294002,a selective inhibitor of PI3K,significantly inhibited the up-regulation of the expression of PI3K,Akt,p-FoxO and CyclinD1,and inhibited the down-regulation of the expression of FoxO,p27^KIP1,Bim by Siwu decoction.Conclusion:Siwu decoction can promote the proliferation of OP9 cells and inhibit its apoptosis by regulating PI3K-Akt-FoxO signaling pathway.