电针对失坐骨神经大鼠腓肠肌细胞凋亡及相关蛋白的影响  

Effects of Electroacupuncture on Apoptosis-related Protein Expression of Gastrocnemius Muscle Cells in Rats with Denervated Sciatic Nerve

作　　者：高睿琦 唐成林[1] 黄思琴[1] 曹净[1] 郭全虎[1] 张毅[1] 田源[1] 袁海洲[1] 赵丹丹[1] 罗翱[1] 张安宁[1] 

机构地区：[1]重庆医科大学中医药学院,重庆400016

出　　处：《针刺研究》2017年第4期302-307,共6页Acupuncture Research

基　　金：国家自然科学基金(No.81273870);重庆卫计委中医药科研项目(No.ZY 20150249)

摘　　要：目的:观察电针疗法对失坐骨神经大鼠腓肠肌细胞凋亡及相关蛋白的影响,探讨电针对失坐骨神经骨骼肌萎缩的延缓效应及其可能机制。方法:SD大鼠随机分为假手术组、模型组和电针组,每组21只。采用切断坐骨神经法制备腓肠肌萎缩大鼠模型。电针组电针患侧"足三里""承山"穴,每日1次,每次10min,各组分别在术后7、14、21d取材。TUNEL法检测腓肠肌细胞凋亡,Western blot法检测Bcl-2、Bcl-2相关的X蛋白(Bax)、细胞色素C(Cyt-C)以及天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)的蛋白表达,RT-PCR法检测腓肠肌Caspase-3mRNA表达。结果:模型组各时点细胞凋亡指数均高于假手术组(P<0.05),电针组的细胞凋亡指数显著低于模型组(P<0.05)。模型组各时点腓肠肌Bcl-2蛋白表达较假手术组显著降低(P<0.05),Bax、Cyt-C以及Caspase-3蛋白表达均明显升高(P<0.05)。术后14、21d,电针组与模型组比较,Bcl-2蛋白表达显著升高(P<0.05),Bax、Cyt-C及Caspase-3蛋白表达则明显降低(P<0.05)。各组大鼠腓肠肌的Caspase-3mRNA表达与蛋白表达结果基本一致。结论:电针能够促进腓肠肌Bcl-2的表达并抑制Bax、Cyt-C和Caspase-3的表达,从而抑制肌细胞的凋亡,这可能是电针延缓失神经性骨骼肌萎缩的机制之一。Objective To observe the anti-apoptosis effect of electroacupuncture（EA）on gastrocnemius muscle cells in rats with denervated sciatic nerve,so as to explore its possible mechanisms underlying delaying atrophy of skeletal muscle.Methods Sixty-three male Sprague-Dawley rats were randomly divided into sham operation group,model group,and EA group,and then further divided into 3subgroups in each group（n=7/subgroup）.Gastrocnemius muscle atrophy model was established by transecting the sciatic nerve of rat.EA（5Hz,1.5mA）was applied to ＂Zusanli＂（ST 36）and ＂Chengshan＂（BL 57）acupoints at the affected side for 10 min,once a day for 1,2,3 weeks,respectively.The gastrocnemius muscles were sampled on the 7～（th）,14 ～（th）,21～（th） d after modeling,separately.The apoptotic cells were detected by TUNEL staining.Bcl-2,Bax,Cyt-C and Caspase-3 protein expressions were determined by Western blot.RT-PCR was used to check Caspase-3 gene expression.Results Compared with the sham operation group,the cell apoptotic index in gastrocnemius was markedly higher,gastrocnemius Bcl-2 protein expression was markedly down-regulated,and gastrocnemius Bax,Cyt-C and Caspase-3 protein expressions were considerably up-regulated in the model group at all time-points（P〈0.05）.Compared with the model group,the cell apoptotic indexes in gastrocnemius were significantly lower in the three EA subgroups（P〈0.05）.Bcl-2 protein expressions were markedly up-regulated while Bax,Cyt-C and Caspase-3 protein expressions were significantly down-regulated in the EA group 14,21 days after modeling compared with the corresponding model subgroups（P〈0.05）.The changes of Caspase-3 mRNA expression levels of gastrocnemius in all groups were similar to those of Caspase-3 protein expressions.Conclusion Electroacupuncture intervention can effectively increase Bcl-2 expression and decrease Bax,Cyt-C and Caspase-3 expression in gastrocnemius muscle,and consequently reduce the apoptosis of muscle cell,which may contribute

关 键 词：失坐骨神经 细胞凋亡 电针 CASPASE-3 

分 类 号：R245.97[医药卫生—针灸推拿学]