The Role Of Akt/GSK3?/?-catenin Pathway In BMSCs In The Development Of Alcohol-induced ONFH

OBJECTIVES: Decreased osteogenesis is one of the important mechanisms of alcohol-induced necrosis of the femoral head.In this study,we investigated whether a novel small molecule drug targeting Akt / GSK3? / ?-catenin pathway in BMSCs can be used in the treatment and prevention of alcoholinduced ONFH by a variety of in vitro and in vivo detection methods.In-depth discussion of the molecular mechanism.METHODS:Inhibition of Akt / GSK3? / ?-catenin pathway in BMSCs was rescued in a series of in vitro experiments by Akt agonist SC79 and p53 inhibitor PFT?,and by a variety of in vivo studies,explore whether it can save the BMSCs caused by ethanol decline in osteogenic differentiation,and the occurrence of alcohol-induced ONFH.RESULTS:(1)Western blot showed that the inhibitory effect of ethanol on osteogenic differentiation of mesenchymal stem cells was mediated through the Akt / GSK3? / ?-catenin pathway.(2)Ethanol can significantly inhibit the phosphorylation level of Akt-Ser473,but no significant change in the phosphorylation level of Akt-Thr308.(3)Ethanol can induce the activation of p53 in BMSCs,activate GSK3?,and decrease the nuclear translocation of ?-catenin,resulting in the decrease of osteogenic differentiation ability of BMSCs.(4)It was found by q PCR,Western blot and alizarin red staining that Akt agonist SC79 and p53 inhibitor PFT? could rescue the inhibitory effect of ethanol on Akt / GSK3? / ?-catenin pathway,Osteogenic differentiation of mesenchymal stem cells.(5)In vivo studies,SC79 and PFT? can alleviate and prevent the occurrence of alcohol-induced ONFH by enhancing osteogenic ability.CONCLUSIONS:(1)Ethanol can inhibit the GSK-3? / ?-catenin signaling pathway by activating p53,eventually inhibiting the osteogenic differentiation of MSCs,leading to alcohol-induced ONFH.As a new and selective p53 inhibitor,PFT? can prevent alcohol-induced ONFH.(2)The phosphorylation of Akt-Ser473 in Akt / GSK3? / ?-catenin signaling pathway was related to the inhibition of alcohol induced osteogenesis,whereas Akt selective agonist SC79 could rescue the osteogenic differentiation of BMSCs,and the occurrence of alcohol-induced ONFH.