Experimental Study On The Effect Of Electroacupuncture At Zusanli On PGC-1α/SIRT3 Signal Pathway In Skeletal Muscle Of Rats With Spleen-qi Deficiency

Purpose:In this study,the rat model of spleen-qi deficiency was duplicated by the combined factors of excessive fatigue and inadequate diet.By observing the expression of PGC-1a/SIRT3 signaling pathway related molecules in skeletal muscle of rats with spleen-qi deficiency by electroacupuncture Zusanli(ST36),the regulation mechanism of Zusanli(ST36)on spleen-qi deficiency syndrome was explored from the perspective of oxidative stress,and the scientific connotation of “spleen governing muscle” was partly explained.Material and method:Thirty-two male SD rats were randomly divided into 4 groups:blank group,spleen-qi deficiency group,Zusanli(ST36)group and non-acupoint group,with 8 rats in each group.All rats were housed in an SPF animal experimental center.A rat model of spleen-qi deficiency syndrome was established by the combination of excessive fatigue and irregular diet.After successful modeling,electro-acupuncture was applied to Zusanli(ST36)and non-meridian acupoints on both sides of rats respectively,once a day,20 minutes each time for 7 days.The blank group and spleen-qi deficiency group were fixed on the operating table for 20 minutes without any other treatment.The mRNA expression levels of PGC-1alpha、ERRalpha、SIRT3、and SOD2 in rat skeletal muscle were detected by real-time PCR.The skeletal muscle PGC-1alpha、ERRalpha、SIRT3、SOD2 were detected by Western blot.Results:1.Observation of general state of rats in each groupThe rats in the blank group were strong in body,shiny in fur,high in excitement and normal in stool.The rats in the spleen-qi deficiency group were thinner,fed less,lost weight,lazy,withered fur and loose stool.After electroacupuncture at Zusanli(ST36)point,the rats’ food intake increased,the body weight increased slightly,the activity increased,the hair loss decreased,and the stool condition improved.The general state of the non-acupoint group rats did not improve significantly compared with the spleen-qi deficiency group.2.Grasping strength comparison of rats in each groupThere was no significant difference in the grip force of rats in each group before the modeling(P>0.05);after the modeling,the grip strength of rats in spleen-qi deficiency group,Zusanli(ST36)group and non-acupoint group decreased significantly compared with the blank group(P<0.05);after the intervention of electroacupuncture at Zusanli(ST36)point,the grip strength of rats in Zusanli(ST36)group increased significantly compared with that in spleen-qi deficiency group(P<0.05),but there was no significant difference between non-acupoint group(P>0.05).3.Comparison of the mRNA expression of PGC-1alpha,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in each groupCompared with the blank group,the mRNA expression of PGC-1alpha,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in spleen-qi deficiency group and non-acupoint group decreased significantly(P<0.05);compared with spleen-qi deficiency group,the mRNA expression of PGC-1alpha,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in Zusanli(ST36)group increased significantly(P<0.05),while there was no significant difference in the non-acupoint group(P>0.05).4.Comparison of the protein expression of PGC-1alpha,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in each groupCompared with the blank group,the protein expression of PGC-1a,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in the spleen-qi deficiency group and non-acupoint group decreased significantly(P<0.05);compared with the spleen-qi deficiency group,the protein expression of PGC-1alpha,ERRalpha,SIRT3 and SOD2 in skeletal muscle tissue of rats in Zusanli(ST36)group increased significantly(P<0.05),but there was no significant difference in the non-acupoint group(P>0.05).Conclusion:1.Electroacupuncture at Zusanli(ST36)can regulate the antioxidant capacity of skeletal muscle of rats with spleen-qi deficiency and improve the symptoms of spleen-qi deficiency and fatigue.2.The antioxidant ability of electroacupuncture at Zusanli(ST36)on skeletal muscle of ratswith spleen-qi deficiency is closely related to PGC-1a/SIRT3 signaling pathway.