电针“委中”大鼠血清对多裂肌卫星细胞成肌分化因子和周期蛋白表达的影响

Effects of Electroacupuncture "Weizhong" Serum on MYOD and Cyclin Expression in Satellite Cells of Polyfidus Muscle

目的观察电针"委中"大鼠血清对大鼠腰多裂肌卫星细胞成肌分化因子MyoD、周期蛋白CDK4、P57和CyclinD表达的影响,从细胞周期角度分析电针"委中"血清促进损伤多裂肌再生修复的起效机制。方法SD大鼠随机分为空白组、模型组和电针"委中"组,每组8只。通过肌肉注射0.5%布比卡因制备多裂肌损伤模型,电针组选取"委中"穴进行电针治疗,每日1次,每次20 min,第4日收集血清。培养原代大鼠腰多裂肌卫星细胞,随机分为空白血清组、模型血清组、电针"委中"血清组、抑制剂血清组(电针"委中"血清中加PI3K抑制剂LY294002),以对应血清分别培养1 d,免疫印迹法检测肌卫星细胞MyoD、CDK4、P57和CyclinD蛋白表达。结果造模后,各血清组MyoD、CDK4和P57表达水平均显著升高(P <0.01);与模型血清组对比,电针"委中"血清组MyoD、CDK4升高(P <0.05或P <0.01)而P57显著降低(P <0.01);与电针"委中"血清组对比,加入LY294002抑制剂后MyoD、CDK4降低(P <0.05或P <0.01)而P57显著升高(P <0.01)。与空白血清组对比,电针"委中"血清组CyclinD升高(P <0.05),其他各组对比差异无统计学意义(P> 0.05)。结论电针"委中"血清可通过上调MyoD、CDK4的表达,并下调P57的表达,加速肌卫星细胞增殖的周期性进程,促进损伤多裂肌的良性修复。

Objective: To observe the effect of electro-acupuncture "Weizhong" serum on the expression of MyoD,CDK4,P57 and Cyclin D in rat lumbar multifidus muscle satellite cells,and to analyze the mechanism of electro-acupuncture "Weizhong" serum promoting the regeneration and repair of injured multifidus muscle from the perspective of cell cycle. Methods: SD rats were randomly divided into the blank group,the model group and electro-acupuncture "Weizhong" group,with 8 rats in each group. The model of polyfid muscle injury was established by intramuscular injection of 0.5% bupivacaine. The electro-acupuncture group selected "Weizhong" point for electro-acupuncture treatment,once a day,20 minutes a time,and collected serum on the fourth day. The primary rat lumbar multifidus muscle satellite cells were cultured and randomly divided into blank serum group,model serum group,electro-acupuncture "Weizhong" serum group and electro-acupuncture "Weizhong" plus LY294002 inhibitor serum group. The corresponding serum was cultured for 1 day. The expression of MyoD,CDK4,CyclinD and P57 protein in muscle satellite cells was detected by Western blot. Results: The expression levels of MyoD,CDK4 and P57 in serum groups were significantly increased after modeling(P < 0.01);compared with model serum group,MyoD and CDK4 in electroacupuncture "Weizhong" serum group increased(P < 0.05 or P < 0.05) and P57 decreased significantly(P < 0.01);compared with electroacupuncture "Weizhong" serum group,MyoD and CDK4 decreased(P < 0.05 or P < 0.01) and P57 increased significantly after adding LY294002 inhibitor(P < 0.01). Compared with the blank serum group,CyclinD in the serum group of electro-acupuncture "Weizhong" increased(P < 0.05),but there was no significant difference among the other groups(P > 0.05). Conclusion: Electroacupuncture "Weizhong" serum can accelerate the cyclical process of muscle satellite cell proliferation by up-regulating the expression of MyoD and CDK4 and down-regulating the expression of P57,and promote the benign repair of damaged polyfidus muscle.