Functional Verification Of The Target Genes(Chrnal And Ipo5)Associated With Blood Flow Restricted Exercise In A Rat Model Of Skeletal Musclar Myopachynsis

Objective To verify the function of the target genes(Chrnal and Ipo5)associated with blood flow restricted exercise(BFRE)in a rat model of skeletal musclar myop achynsis in vivo and in vitro by using RT-PCR and Western blots,to investigate the e ffect of Myo statin mRNA expression under the influence of the target genes’ siRNA(small interfering RNA)or their over-expressed plasmid,confirming their closely asso ciated with muscle atrophy and hypertrophy.Methods Rat muscle cell line L6 was used in the functional verification of the target genes in vitro,including four groups:the normal group,the gene silencing group,the gene over expression group,the gene rescue group;Primitive cells of rat skeletal muscle was used in the functional verification of the target genes in vivo,including five groups:the normal group,the model group,the gene silencing group,the gene over expression group,the gene rescue group;After transfection was optimized,the extracted total RNA and the total protein at 48h after transfection,the real-time quantitative PCR and Western blots were used to detect the expresstion changes of chrnal,Ipo5 and Myostatin.Results The over expression of Chrnal can downregulate the expression of Myostatin in rat muscle cell line L6,which can also be confirmed by Western blots.Chrnal siRNA can obviously increase the expression of Myostatin in rat muscle cell line L6,which can also be confirmed by the result of Western blots.The rescued group had a lower expression of Myostatin compared with the siRNA interference group.Chmal can downregulate the expression of Myostatin in some way,so as to increase muscle protein synthesis.The over expression of Ipo5 can raise the expression of Myostatin in rat muscle cell line L6,which can also be confirmed by the result of Western blots.Ipo5 siRNA of can obviously decrease the expression of Myostatin in Rat muscle cell line L6,which can also be confirmed by the result of Western blots.The rescued group had a higher expression of Myostatin compared with the siRNA interference group.In the other words,Ipo5 can raise the expression of Myostatin in some way,so as to promote muscle atrophy.Conclusion Chrnal,Ipo5 can both affects the expression of Myostatin,confirming its closely associated with muscle atrophy and hypertrophy and establishing a foundat ion for gene therapy for muscle atrophy.