Effect of decoction for replenishing Qi and bonesetting on bone metabolism in rats with steroid-induced osteonecrosis of femoral head via regulating PI3K/Akt signaling pathway
Objective It is to investigate the effect of decoction for replenishing Qi and bonesetting(DRQB)on bone metabolism in rats with steroid-induced osteonecrosis of femoral head(SONFH)via regulating the phosphoinositide 3-ki-nase(PI3K)/protein kinase B(Akt)signaling pathway.Methods Fifty SD male rats were randomly divided into normal group,model group,DRQB low-dose group,DRQB high-dose group and DRQB+LY294002 group,with 10 rats in each group.The rats of all group except for the normal group were injected with prednisolone acetate intramuscularly in the right buttock for 4 weeks to establish model of SONFH.At the end of modeling,the rats in the low and high dose groups of DRQB were gavaged with 0.5 mL and 1 mL of DRQB,respectively,and the rats in the DRQB+LY294002 group were ga-vaged with 1 mL of DRQB and intraperitoneally injected with 6 mg/kg of LY294002,while the rats in the normal group and model group were given the same volume of normal saline by gavage,all once daily,continuously treated for 8 weeks.The serum levels of interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),calcium,phosphorus,os-teoprotegerin(OPG),osteocalcin(OCN)and alkaline phosphatase(ALP)of the rats were determined by ELISA,the morphology of femoral head was detected and observed by Micro-CT,the pathologic morphology of femoral head tissues was observed by HE staining,the osteoclastogenesis was observed by TRAP staining,and the expressions of bone-specific tran-scription factor 2(Runx2),bone morphogenetic protein 2(BMP-2),Cleaved-Caspase-3,PI3K,and Akt proteins in the femoral head tissues were detected by Western blot.Results Compared with the normal group,the serum levels of IL-4,1L-6,TNF-α,OPG and ALP,the trabecular bone resorption(Tb.Sp),percentage of empty lacunae,number of osteoclasts and relative expressions of Cleaved-Caspase-3 protein in femoral head tissue were significantly increased in the rats of the model group(all P＜0.05),while the serum levels of calcium,phosphorus and OCN,bone volume fraction(BV/TV),number of trabeculae(Tb.N),trabecular thickness(Tb.Th),relative expressions of Runx2,BMP-2,p-PI3K/PI3K and p-Akt/Akt proteins in femoral head tissue were significantly decreased(all P＜0.05),the femoral head was greatly dam-aged,and the spatial structure of the bone trabeculae was disordered.Compared with the model group,the serum levels of IL-4,IL-6,TNF-a,OPG and ALP,Tb.Sp,percentage of empty lacunae,number of osteoclasts and relative expressions of Cleaved-Caspase-3 protein in femoral head tissue were significantly decreased in the rats of the low and high dose groups of DRQB(all P＜0.05),while the serum levels of calcium,phosphorus and OCN,(BV/TV,Tb.N,Tb.Th,relative ex-pressions of Runx2,BMP-2,p-PI3K/PI3K and p-Akt/Akt proteins in femoral head tissue were significantly increased(all P＜0.05),the damage of the femoral head was alleviated,and the spatial structure of bone trabeculae was improved;LY294002 could attenuate the improvement effect of DRQB on the above indexes,and the differences in all indexes were statistically significant when comparing with those of the high-dose group of DRQB(all P＜0.05).Conclusion Decoction for replenishing Qi and bonesetting can improve bone metabolism,reduce the levels of inflammatory factors,decrease the number of osteoclasts,and attenuate the necrosis of the femoral head in rats with hormonal necrosis of femoral head,and the mechanism may be related to the activation of the PI3K/Akt signaling pathway.
hormone induced necrosis of femoral headdecoction for replenishing Qi and bonesettingphosphoinosit-ide 3-kinaseprotein kinase Bbone metabolism
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