| Objective:In this study,we observed the effects of massage manipulation on autophagy,apoptosis,and oxidative stress related factors in the repair of acute blunt injury of skeletal muscle in rats.To explore the possible biological therapeutic mechanism of massage therapy for acute skeletal muscle injury.Methods:Forty-two adult male Sprague-Dawley rats were randomly divided into three groups: group A(Normal group,n=6),group B(Model group,n=18)and group C(Treatment group,n=18).Group B and group C were randomly divided into three subgroups: 3 days group,9 days group and 16 days group.The acute blunt injury model of gastrocnemius muscle was established by using self-provided instruments.The treatment group was treated with massage after 48 hours of model making,once a day for 15 minutes each time.The injured gastrocnemius muscle was taken out in each group at 3 days,9 days and 16 days after cardiac blood collection.The morphological changes of gastrocnemius were observed by HE staining,the expression of LC3,Beclin1,P62 in gastrocnemius was detected by Immunohistochemistry and Western blot,the level of apoptosis in gastrocnemius muscle was detected by TUNEL,and the expression of Bax,Bcl-2,Cyt-C,Caspase-3 in gastrocnemius muscle were detected by Western blot,the serum content of SOD and MDA were detected by WST-1 and TBA,the serum content of TNF-??IL-1??IL-6?CRP?PGE2 were detected by ELISA.Results:During the repair of skeletal muscle injury,The results of HE staining showed that the morphology of the cells in the model group was more than that in the normal group,and the nuclei of the myogenic cells appeared at the day of 9 and 16,the recovery of the cells in the treatment group was better than that in the model group,and the number of neonatal muscle cells in the treatment group was more than that in the normal and model group at each time point,the shape is more normal.The results of immunohistochemistry showed that the expression of autophagy associated protein LC3-II,Beclin1,P62 of gastrocnemius muscle in group B and group C was significantly raised contrast with group A,the expression of LC3-II and Beclin1 was significantly higher in group C than that in group B at different time points(P< 0.05,P < 0.01,P< 0.001),the expression of P62 was significantly lower in group C than that in group B at different time points(P< 0.05,P < 0.01).The results of Western blot showed that the expression of LC3-II/I and Beclin1 in gastrocnemius muscle of group B was significantly reduced contrast with group A at each time point,and the expression of P62 in group B was significantly raised contrast with group A at each time point.At the same time point,the expression of LC3-II/I and Beclin1 of gastrocnemius muscle in group C was significantly higher than that in group B(P< 0.01).the expression of P62 in group C was significantly lower than that in group B(P< 0.01).The results of immunofluorescence showed that the apoptosis index of group B and group C was significantly raised contrast with group A at each time point.the apoptotic index of group C was significantly lower than that of group B(P<0.05,P<0.01).The results of Western blot showed that the expression of apoptosis-related protein Bax,Bcl-2,Cyt-C,Caspase-3 in group B and group C was significantly raised contrast with group A at each time point,the expression of Bax,Cyt-C,Caspase-3 in group C was significantly lower than that in group B(P<0.05,P<0.01,P<0.001),and the expression of Bcl-2 in group C was significantly higher than that in group B(P < 0.01,P<0.001).The results of WST-1 and TBA showed that the serum content of SOD and MDA in group B were significantly raised contrast with those in group A at each time point after injury.The serum content of SOD in group C was significantly higher than that in group B(P< 0.05),the serum content of MDA in group C was significantly lower than that in group B(P< 0.05).The results of ELISA showed that the serum content of proinflammatory factor in group B was significantly raised contrast with that in group A at each time point after injury,but in group C they were significantly lower than that in group B(P< 0.01,P< 0.05).Conclusion:Massage can effectively promote the repair of acute skeletal muscle contusion,and its mechanism may be related to the increase of autophagy activity of muscle tissue cells and inhibition of apoptosis of normal cells.It may also be related to the reduction of oxidative stress and the decrease of inflammatory response of injured body by massage,thus accelerating the repair speed of skeletal muscle injury. |
PDF Full Text Request