| ObjectiveThe purpose of this study was to examine the effects of massage on expressions of plasma membrane repair proteins in rats with skeletal muscle acute contusion, and the effects of massage on muscle biomechanical properties following contusion were studied too,so as to explore the possible mechanism of massage therapy in repair of skeletal muscle plasma membrane injury.Methods1.Forty-two SD rats were randomly divided into control group(n=6)and model group. Acute contusion on the right gastrocnemius muscle were made in model group and thirty-six rats of the model group were divided randomly into three groups with 7d,14 d,21d of contusion respectively,each group contains natural recovery group(n=6) and massage therapy group(n=6). Massage was given to rats in the massage therapy group on local damage surface after 48 h injury while other groups without anytreatment. HE staining to observe the pathologic changes of the muscle tissue. Double-labeling immunofluorescence staining to observe the co-localization of dysferlin with annexin A1 and Western-blot analysis was used to detect the protein expressions of dysferlin、 annexin A1 and MG53.2.Twenty-eight SD rats were randomly divided into control group( n=4) and model group. Acute contusion on the right gastrocnemius muscle were made in model group and eighteen rats of the model group were divided randomly into three groups with 7d,14 d,21d of contusion respectively,each group contains natural recovery group(n=4) and massage therapy group(n=4). The biomechanical performance of the injured gastrocnemius were tested in each group.Results1. HE staining showed that there were no inflammatory cells infiltration and cells morphology were normal in control group. At 7d,natural recovery group and massage treatment group had obvious cells morphological deformation, cells disintegration and atrophy, the inflammatory infiltration was increased significantly in cell gap, there were no significant differences between the two groups. At 14 d, cells in natural recovery group were disintegrated, atrophy, cell gap was filled with connective tissue, and cells massage treatment group atrophy was alleviated, cells arranged relatively closely, new muscle fibers and smallvascular were found. At 21 d, new muscle cells and vascular cells natural were restoration, connective tissue staggered obviously in natural recovery group, and the massage treatment group with a small amount of connective tissue filling, but the cell shape had a significant recovery, cells arranged closely and tightly, angiogenesis significantly.2.Double-labeling immunofluorescence staining revealed that dysferlin and annexin A1 were distributed in plasma membrane or in control group and had a lower expressions without co-expressions. After injuried they were diffusedly distributed in cells especially in 7d and reduced with times, the expressions of two proteins in massage therapy group were lower than natural recovery group at the same time points and had more co-expressions in plasma membrane.2.Western blot showed a significant upregulation of three proteins in model groups than control group,the expressions of the proteins in natural recovery groups were reduced with time and the differences among the groups at 7d, 14 d and 21 d were statistically significant(P<0.05), and the massage therapy groups had a same tendency(P<0.05).The expressions of dysferlin, annexin A1 and MG53 of massage therapy groups had a lower expressions than natural recovery group at 21d(P<0.05), as well as the expressions of annexin A1 and MG53 at 14 d, 21d(P<0.05)and 7d, 14 d,21d(P<0.05)respectively.2.Tonic contraction tension tests showed an increased tension in eachgroups compared with control group(P<0.05). With time went by, the tonic contraction tension natural recovery group and the massage therapy group were decreased gradually. At 7d and 14 d, the tonic contraction tension had no significant difference between massage treatment group and natural recovery group(P>0.05). At 21 d, the tonic contraction tension of massage treatment group closed to control group and had a significant difference compared with the natural recovery group(P<0.05).Conclusion :Massage may played a role on the regulations of membrane repair proteins after skeletal muscle acute contusion in the experiment.Co-expressions of dysferlin and annexin A1 as well as the reduction of the proteins expressions of dysferlin, annexin A1 and MG53 in massage therapy group may be related with inflammatory response and suggesting that massage might promoting the repair of membrane healing. Besides,massage plays a beneficial role in decreasing the tonic contraction tension following contusion injury and improve the quality of repair in injured skeletal muscle. |
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