Effect of Regulation of PI3K/AKT/FoxO1 Signaling Pathway by Gubi Tongxiao Granules on the Differentiation of Human Bone Mesenchymal Stem Cells
Objective To investigate the protective effect of Gubi Tongxiao granules on steroid-induced necrosis of femoral head(SANFH).Methods Femoral head and bone marrow tissues of patients with SANFH and femoral neck fracture(FNF)undergoing hip replacement were collected and studied.The expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),FoxOl and cholesterol regulatory binding protein(SREBP)were detected in bone tissue.Human bone marrow mesenchymal stem cells(hBMSCs)were isolated and cultured.The effect of the drug-containing serum of Gubitongxiao granule on the proliferation of hBMSCs was detected by CCK-8 method.Oil red O and alizarin red staining were used to observe the adipogenic and osteogenic differentiation of cells.The expression of peroxisome proliferator-activated receptor γ(PPARy),CCATT enhancer binding protein α(C/EBPα),Runt-associated transcription factor 2(Runx2),alkaline phosphatase(ALP)mRNA and pathway protein were analyzed by RT-qPCR and Western blot.Results Western blot results showed that the expression of PI3K,AKT and SREBP protein in femoral head tissue of SANFH patients was significantly increased compared with that of normal group,while FoxOl expression was significantly decreased(P＜0.05);Different volume fractions of Gubi Tongxiao granules could promote the proliferation of hBMSCs,especially in 5％group after 48h.Compared with the model group,5％drug-containing serum could significantly reduce the lipid droplet coloring in the cytoplasm and increase the orange-red mineralized nodules in the calcium salt deposit,down-regulate the mRNA expression levels of PPARγ and C/EBPα,and up-regulate the mRNA expression levels of Runx2 and ALP(P＜0.05).In addition,after the intervention of drug-containing serum,the expression of FoxO1 protein in cells was significantly increased,and the expression of AKT,p-FoxO1 and SREBP protein was decreased(P＜0.05).Conclusion After the intervention of Gubi Tongxiao granules,the lipid differentiation of hBMSCs decreased and the osteogenic differentiation increased,which can effectively improve the lipid deposition in glucocorticoid environment,and the mechanism may be related to the regulation of PI3K/AKT/FoxO1 signaling pathway.
Steroid-induced avascular necrosis of femoral headGubi Tongxiao granulesOsteogenic and adipogenic differentiationPhosphorylationPI3K/AKT/FoxO1 signaling pathway
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