按摩对大鼠骨骼肌急性钝挫伤后膜修复蛋白dysferlin和annexin A1表达的影响

关键词：dysferlin  annexin A1  急性钝挫伤  按摩  膜修复

Keywords:dysferlin  annexin A1  acute contusion  massage  membrane repair

摘要：

      摘要 目的：研究大鼠骨骼肌急性钝挫伤后，按摩对细胞膜特异性修复蛋白dysferlin和annexin A1表达的影响，探讨按摩促进骨骼肌急性损伤修复的可能机制。 方法：42只成年SD雄性大鼠随机抽取6只作为正常对照组，其余36只为模型组。模型组在建立右下肢急性腓肠肌钝挫伤模型后，随机分为自然恢复组和按摩治疗组，根据治疗时间分为损伤后7d、14d、21d三个时间点，每个时间点6只。正常对照组和自然恢复组不做治疗；按摩治疗组在建模后48h开始每天于损伤局部行按摩治疗。免疫荧光双标法标记dysferlin和annexin A1，运用激光共聚焦显微镜摄片，观察其在细胞内分布及表达；运用蛋白质印迹法检测其表达量。 结果：激光共聚焦显微镜结果显示:正常肌细胞中两种荧光信号主要分布于近细胞膜周围，强度较弱，无共表达现象；模型组各时间点，两种荧光信号强度均明显增加，胞浆内也有弥散性分布，随时间的延长，信号强度逐渐减弱，细胞膜共表达现象增加；其中按摩治疗组荧光信号强度较同时期自然恢复组减弱明显，细胞膜上共表达现象增多。蛋白质印记结果显示:模型组各时间点较正常对照组，dysferlin和annexin A1蛋白表达升高，差异显著;随着时间的延长，模型组蛋白表达量均下降，自然恢复组和按摩治疗组在损伤后14d和21d比较，差异均有显著性意义（P<0.05）；损伤后21d，自然恢复组较按摩治疗组dysferlin表达量比较，差异有显著性意义（P<0.05）；损伤后14d和21d，自然恢复组较按摩治疗组annexin A1表达量比较，差异有显著性意义（P<0.05）。 结论：这可能是其促进肌细胞膜修复相关机制之一。

Abstract:

      Abstract Objective：To investigate the effect of massage on the expressions of cytomembrane repair proteins dysferlin and annexin A1 in rats with acute skeletal muscle contusion and to explore the possible mechanism of massage therapy on repairment of skeletal muscle injury. Method：Forty-two SD rats were randomly divided into control group（n=6）and model group（n=36）. Acute contusion on the right gastrocnemius muscle were made in model group and rats of the model group were divided randomly into three groups with 7, 14, 21 days treatment respectively. Each point in time contains natural recovery group（n=6） and massage therapy group（n=6）. Massage was given to rats in massage therapy group on damaged surface daily since 48 h after injury while no treatment was given to other groups. The distribution and quantity of protein expressions of dysferlin and annexin A1 were taken by double-labeling immunofluorescence staining and Western-blot analysis. Result：In normal cell, double-labeling immunofluorescence staining revealed that dysferlin and annexin A1 distributed on cytomembrane with lower expressions and no co-expressions. In model group and diffusedly distributed in cells, But it reduced with time. 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 on cytomembrane .Western blot showed that upregulation of two proteins in model groups significantly more than in control group. The expressions of two proteins in natural recovery group and massage therapy group reduced with time and the differences among two groups were statistically significant（P<0.05）.The massage therapy group had lower expressions of dysferlin than natural recovery group at 21d（P<0.05）,as well as lower annexin A1 at day 14 and day 21（P<0.05）. Conclusion：Massage can reduce the expressions of dysferlin and annexin A1 at cytomembrane of acute contusion muscle. It may be related to the mechanism of cytomembrane repair.