Effects Of Electroacupuncture Combined With Treadmill Training On Acute Skeletal Muscle Injury Repair In Rats With Neuromuscular Junction Associated Proteins Nestin Cdk5 Epsilon-nAChR

Objective:To observe the effect of treadmill and electro-acupuncture interventions on rats with skeletal muscle acute contusion,so as to reveal the role of?-nAChR?Cdk5 and nestin on the recovery of NMJ.Method:60 adult male SD rats were randomly divided into 2 groups : control group(n=6)and natural recovery group and treatment group.In the natural recovery group and treatment group,the rats who were induced acute injury model of right lower limb were divided randomly into 2groups(n=24): natural recovery group and treatment group.After 48 h,the natural recovery group and treatment group would start Treadmill training schedule,then taken EA with ZuSanli and A Shi xue for once-daily,continuous five days after the intervention,take two days off every week.The testing was made at different time points,seventh day,fourteenth day,twenty-first day and twenty-eighth days after the rats were injured.The morphological alterations were observed by HE stain;Nestin?Cdk5 and ?-nAChR were tested by Western blot?Immunofluorescence and RT-PCR.Result:1 HE staining showed: In the normal group,the shape of skeletal muscle fibers integral and arranged neatly;at the same point time,the skeletal muscle fibers in natural recovery group;the rupture?denaturation necrosis were worse than normal control group,and there were a few of newborn satellite cell with centronucleus;the skeletal muscle fibers organization structure in intervention group regained more faster than normal control group,the extravasated blood and edema in the orgnazation vanished faster,the newborn muscle fibers were more than control group.2 Immunofluorescence staining:In the natural recovery group and treatment group,nestin showed coexpression with Cdk5 on the cytomembrane;in normal control group.the coexpression fluorescence intensity showed less and less among the 4 observe time poins.3 Western blot showed:In the natural recovery group and treatment group:nestin showed no expression in twenty-eighth days(P >0.05),but showed more expression than natural recovery group in seventh days and fouth days and twenty-first days(P <0.05);Cdk5 expression were more than normol group in seventh day,so did fourteenth days and twenty-first days(P <0.05),at these observe time points.the intervention group expressin less than natural recovery group(P <0.05),but both of group's expression were almost the same in tweenty-eighth days(P >0.05);?-nAChR expression increased with the observe time points(P <0.05),at last,the natural recovery group and treatment group had no difference with normal group in tweenty-eighth days(P >0.05),but treatment group showed more expression than natural recovery group.4 RT-PCR expression showed:In the natural recovery group and treatment group:nestin mRNA showed more expression than natural recovery group in seventh days and fouth days(P <0.05),at last,the natural recovery group and treatment group had no difference with normal group in tweenty-First day and tweenty-eighth days(P >0.05).Cdk5 mRNA expression in the natural recovery group and treatment group were more than normol group in seventh day,so did fourteenth days and twenty-first days(P <0.05),at these observe time points.the intervention group expressin less than natural recovery group(P <0.05),but both of groups' expression were almost the same(P >0.05).The expression of ?-nAChR mRNA n the natural recovery group and treatment group were less than inormol group in seventh day,so did fourteenth days and twenty-first days(P <0.05),at these observe time points.the intervention group expressin more than natural recovery group(P <0.05),in tweenty-eighth days,the natural recovery group and treatment group had no difference with normal group(P >0.05),but treatment group showed more expression than natural recovery group.Conclusion: Electroacupuncture combined with treadmill exercise contribute to promote the recovery of NMJ on the remodeling of skeletal muscle after blunt trauma.The mechanism may be related to improving the expression of ?-nAChR?nestin and restrains Cdk5 expression at the injury site.