强力益气颗粒对H2O2损伤大鼠L6骨骼肌成肌细胞的作用及机制研究OA

Objective By observing the effect of Qiangli Yiqi Granule on myoblasts of L6 skeletal muscle in rats injured by H2O2,the possible mechanism of action of MED 13/Nurr1/GLUT-4,a member of nuclear receptor 4 family,in MG was discussed.Methods After the L6 skeletal muscle myoblasts were isolated and cultured,they were divided into six groups:the normal serum group,the model group,the blank serum group,and the high,medium and low dose groups of serum containing Qiangyiqi Granule.Except for the normal serum group,all the other groups used H2O2 to cause oxidative damage.The normal serum group and the model group were cultured in FBS complete medium respectively.The blank serum group was incubated with normal rat serum(excluding the influence of PBS on cells),while the high,the medium and the low dose groups of Qiangyiqi medicated serum were incubated with rat medicated serum in corresponding dose groups,and the cells were collected after 24 hours of culture.Cell proliferation and apoptosis cycle were detected.Real-time PCR,immunofluorescence and Western blot were used to detect the quantitative,localized and qualitative expressions of Qiangli Yiqi serum on L6 skeletal muscle myofibrils PGC-1α,MEF2,MED13,Nurr1 and GLUT-4 protein.Results H2O2 treatment for 2 hours of the proliferation of myoblasts was significantly inhibited in L6 rats(P＜0.05),and the apoptosis rate was significantly increased(P＜0.05).The medicated serum of Qiangli Yiqi Granule can improve the proliferation rate of myoblasts in L6 skeletal muscles of rats injured by H2O2 and reduce the transfer to green.MED13 gene was significantly up-regulated at transcription level and protein level(P＜0.05).Compared with the model group,the transcription level of Nurr1 gene was decreased(P＜0.05).For PGC-1α,MEF2 and GLUT-4,H2O2 treatment significantly reduced their transcription levels(P＜0.05),and pretreatment of drug-containing serum could reverse this extent(P＜0.05),and at the same time a certain dose dependence was showed.Conclusion The medicated serum of Qiangli Yiqi Granule has a protective effect on L6 skeletal muscle myoblasts injured by H2O2 by increasing the protein levels of MED13,GLUT-4,PGC-1α,MEF2,and decreasing the protein level of Nurr1.It is concluded that the medicated serum of Qiangli Yiqi Granule can improve the glucose metabolism of skeletal muscle,reduce the damage of skeletal muscle cells,and further improve the pathology of impaired function of MG skeletal muscle by interfering with MED13/Nurr1/GLUT4 signaling pathway.