电针对骨骼肌钝挫伤模型大鼠细胞周期素依赖性蛋白激酶5表达的影响
Effects of electroacupuncture on the expression of cyclin-dependent kinase 5 after skeletal muscle contusion
目的:观察电针对骨骼肌钝挫伤大鼠细胞周期素依赖性蛋白激酶5( CDK5)表达的影响,探讨电针治疗骨骼肌损伤、促进肌肉功能恢复的作用机制。方法将32只SD大鼠按照随机数字表法分为正常组(n=4)、模型组(n=4)、自然恢复组(n=12)、电针组(n=12),正常组不做特殊处理,其余各组使用自制重物打击器建立骨骼肌钝挫伤模型。自然恢复组不做电针干预、自然恢复;电针组于造模48 h后开始电针干预,每日1次,每次15 min。模型组于建模24 h后处死,目的在于验证造模成功,自然恢复组、电针组分别于造模后第7、14、21天处死取材,使用苏木精-伊红( HE )染色观察组织形态变化,采用免疫印迹和实时定量荧光PCR分别检测CDK5蛋白及其mRNA的表达情况。结果 HE染色显示,与正常组比较,各组可见大量肌纤维断裂溶解及炎性细胞浸润。电针组与自然恢复组比较,肌卫星细胞增殖更快,新生肌纤维明显增多,修复更为迅速。自然恢复组、电针组两组CDK5蛋白表达量较正常组均升高( P<0.05),且电针组明显低于自然恢复组( P<0.05),PCR检测结果也进一步表明,自然恢复组mRNA表达水平显著高于电针组( P<0.05),且均高于正常组( P<0.05)。结论骨骼肌钝挫伤可使大鼠骨骼肌中CDK5蛋白表达升高,电针促进骨骼肌肌肉功能恢复的机制可能与抑制CDK5蛋白及mRNA的表达水平有关。
更多Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.
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