电针调控miRNA206/HDAC4轴促进损伤多裂肌修复过程中成肌细胞分化的机制  

Regulation of miRNA206/histone deacetylase 4 by electroacupuncture promotes myogenic differentiation during multifidus muscle repair

作　　者：刘通[1] 于佳妮[2] 邹德辉 邝伟川[1] 王小寅[1] 文希[1] 江烨[1] 邱晓佳[1] 曾遥 刘悦[1] Liu Tong;Yu Jiani;Zou Dehui;Kuang Weichuan;Wang Xiaoyin;Wen Xi;Jiang Ye;Qiu Xiaojia;Zeng Yao;Liu Yue(Department of Acupuncture and Rehabilitation,Guangdong Second Hospital of Traditional Chinese Medicine,Guangzhou 510095,Guangdong Province,China;Department of Rehabilitation,Guangdong Hospital of Traditional Chinese Medicine,Guangzhou 510120,Guangdong Province,China;School of Traditional Chinese Medicine,North China University of Science and Technology,Tangshan 063210,Hebei Province,China)

机构地区：[1]广东省第二中医院针灸康复科,广东省广州市510095 [2]广东省中医院康复科,广东省广州市510120 [3]华北理工大学中医学院,河北省唐山市063210

出　　处：《中国组织工程研究》2019年第33期5341-5346,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然基金青年基金项目(81704179),项目负责人:刘通~~

摘　　要：背景:作者前期的动物及细胞实验研究已证明电针可提高损伤腰多裂肌中Pax7 和成肌分化抗原的表达,促进多裂肌的损伤修复。亦有研究证明miRNA206 促进成肌分化主要是通过抑制组蛋白去乙酰化酶4 实现的。目的:观察电针对布比卡因致大鼠腰多裂肌损伤修复过程中miRNA206/组蛋白去乙酰化酶4 表达的影响。方法:实验方案经广东省第二中医院动物实验伦理委员会批准(批准号为048604)。将30 只雄性SD 大鼠随机分为对照组、模型组、电针组,每组10 只。模型组、电针组采用0.5%布比卡因肌内注射复制大鼠多裂肌损伤模型;对照组予注射等量生理盐水。对照组与模型组不进行针刺干预,电针组予针刺双侧委中穴、肾俞穴,针刺后连接电针,波形选用疏密波,电针频率采用2 Hz/10 Hz,电流强度选择1 mA,持续治疗20 min,每天治疗1 次,共治疗7 d。干预7 d 后,通过苏木精-伊红染色观察损伤部位多裂肌形态学变化,采用qRT-PCR检测多裂肌中miRNA206 的表达量,Western-blot 检测多裂肌中成肌分化抗原、Myogenin 及组蛋白去乙酰化酶4 蛋白的表达。结果与结论:①苏木精-伊红染色显示,7 d 后,对照组可见视野中骨骼肌纤维大部分排列整齐,无明显破坏及巨噬细胞浸润;模型组视野内可见大量肌纤维被破坏,但出现部分肌纤维修复,巨噬细胞数量仍较多;电针组新生肌纤维较多,巨噬细胞较模型组减少;②Western-blot 结果显示,7 d 后,模型组成肌分化抗原、Myogenin、组蛋白去乙酰化酶4 蛋白表达均较对照组升高(P < 0.05 或P < 0.01);电针组成肌分化抗原、Myogenin 蛋白表达较模型组升高(P < 0.01),组蛋白去乙酰化酶4 蛋白较模型组降低(P < 0.01);③qRT-PCR结果显示,7 d 后,模型组miRNA206 表达高于对照组(P < 0.01),电针组miRNA206 表达高于模型组(P <0.05);④结果说明,电针可促进多裂肌损伤后成肌分化,其作用可能是通过提高miRNA20BACKGROUND: Our preliminary studies on animal and cell experiments have shown that electroacupuncture can improve the expression ofPax7 and myogenic differentiation antigen in injured multifidus muscle and facilitate the repair of multifidus muscle injury. Studies have alsoshown that miRNA206 promotes myogenic differentiation mainly through inhibition of histone deacetylase 4.OBJECTIVE: To explore the effect of electroacupuncture on miRNA206/histone deacetylase 4 during muscle regeneration afterbupivacaine-induced multifidus muscle injury in rats.METHODS: The study protocol was approved by the Animal Ethics Committee of Guangdong Second Hospital of Traditional ChineseMedicine with the approval No. 048604. Thirty male Sprague Dawley rats were randomly divided into control group, model group andelectroacupuncture group (n=10/group). Rat models of multifidus muscle injury were established by injecting 0.5% bupivacaine hydrochloridein model and electroacupuncture groups, while normal saline injection was only given in control group. No acupuncture intervention was givenin the control and model groups. Electroacupuncture at bilateral Shenshu (BL23) and Weizhong (BL40) acupoints was given in theelectroacupuncture group. Needles were then stimulated electrically using a Han’s Acupoint Nerve Stimulator for 20 minutes daily for 7continuous days, with a density wave, frequency of 2 Hz/10 Hz and continuous current of 1 mA. At 7 days of intervention, morphologicalchanges of the multifidus muscle was observed using hematoxylin-eosin staining, and qRT-PCR was used to detect miRNA206 expression,and western blot was used to observe the expression of myogenic differentiation antigen, Myogenin and histone deacetylase 4 in themultifidus muscle.RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining indicated that, most of the skeletal muscle fibers in the control group wereneatly arranged at 7 days of treatment, with no obvious damage and macrophage infiltration. The muscle fibers in the model group werelargely damaged, but partia

关 键 词：电针 多裂肌 修复 形态学 成肌分化 miRNA206 组蛋白去乙酰化酶4 成肌分化抗原 MYOGENIN 

分 类 号：R496[医药卫生—康复医学] R454[医药卫生—临床医学]