基于Rho/ROCK信号通路探讨电针任脉、督脉经穴缓解类痛经模型大鼠子宫平滑肌痉挛的机制  

Mechanisms of electroacupuncture relieves uterine smooth muscle spasm in dysmenorrhea rats based on Rho/ROCK signaling pathway

作　　者：路广通 范玺胜 王雪松 左广 张怡 张明健 林杪 刘今 刘君 张俊茶 师旭亮 佘延芬 LU Guang-tong;FAN Xi-sheng;WANG Xue-song;ZUO Guang;ZHANG Yi;ZHANG Ming-jian;LIN Miao;LIU Jin;LIU Jun;ZHANG Jun-cha;SHI Xu-liang;SHE Yan-fen(School of Acupuncture-moxibustion and Tuina,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei International Joint Research Center for Dominant Diseases in Chinese Medicine and Acupuncture,Shijiazhuang 050200;Bejing Jingmei Group General Hospital,Beijing 102399)

机构地区：[1]河北中医药大学针灸推拿学院,石家庄050200 [2]河北省中医针灸优势病症国际联合研究中心,石家庄050200 [3]北京京煤集团总医院,北京102399

出　　处：《针刺研究》2024年第4期367-375,共9页Acupuncture Research

基　　金：国家自然科学基金面上项目(No.81973755);国家重点研发计划项目(No.2022YFC3500601)。

摘　　要：目的:基于Rho/Rho激酶(ROCK)信号通路,探讨电针任、督二脉经穴缓解子宫平滑肌痉挛、改善原发性痛经(PD)的作用机制并观察任、督二脉经穴作用是否存在差异。方法:SD雌性大鼠随机分为盐水组、模型组、任脉组、督脉组及非经非穴组,每组12只。采用苯甲酸雌二醇联合缩宫素(OT)建立类痛经大鼠模型。任脉组电针“气海”“中极”并针刺“关元”,督脉组电针“命门”“腰俞”并针刺“腰阳关”,非经非穴组电针左侧“非穴1”“非穴3”并针刺“非穴2”,每次20 min,1次/d,连续10 d。观察各组大鼠腹腔注射OT(2 U/只)30 min内的扭体反应;多导生理记录仪记录大鼠在体子宫平滑肌电信号,HE染色法观察大鼠子宫组织的病理形态变化;ELISA法检测大鼠子宫组织前列腺素F_(2α)(PGF_(2α))、OT、钙离子(Ca^(2+))的含量,Western blot及荧光定量PCR法检测子宫组织平滑肌22-α(SM22-α)、RhoA、ROCKⅡ蛋白及mRNA表达水平。结果:与盐水组比较,模型组大鼠扭体评分升高(P<0.01),在体子宫平滑肌振幅电压升高(P<0.01),HE染色可见子宫内膜大范围剥脱伴有严重的水肿,且腺体分泌增加,子宫组织中PGF_(2α)、OT、Ca^(2+)含量升高(P<0.01),子宫组织SM22-α、RhoA、ROCKⅡ的蛋白及mRNA的表达升高(P<0.01)。与模型组和非经非穴组比较,任脉组、督脉组扭体评分均降低(P<0.01,P<0.05),子宫平滑肌振幅电压降低(P<0.01),子宫内膜水肿程度较轻,腺体分泌减少,子宫组织中PGF_(2α)、OT、Ca^(2+)的含量降低(P<0.01,P<0.05),子宫组织SM22-α、RhoA、ROCKⅡ的蛋白及mRNA的表达降低(P<0.01,P<0.05)。结论:电针任脉、督脉经穴均可降低类痛经大鼠的扭体评分、子宫平滑肌振幅电压、子宫病理损伤程度,缓解子宫平滑肌的痉挛。其机制可能与通过调控子宫组织PGF_(2α)、OT含量,抑制子宫组织Rho/ROCK信号通路,降低SM22-α、RhoA、ROCKⅡ蛋白与mRNA表达及Ca^(2+)含量有关Objective To investigate the effect of electroacupuncture(EA)on Rho/Rho-associated coiled-coilforming kinases(ROCK)signaling pathway of uterus tissue in rats with dysmenorrhea,so as to explore the underlying mechanism of EA treating primary dysmenorrhea(PD)and uterine smooth muscle spasm,and to observe whether there is a difference in the effect of meridian acupoints in Conception Vessel(CV)and Governer Vessel(GV).Methods Sixty female SD rats were randomly divided into saline,model,CV,GV,and non-acupoint groups,with 12 rats in each group.The dysmenorrhea model was established by subcutaneous injection of estradiol diphenhydrate combined with intraperitoneal injection of oxytocin(OT).EA(2 Hz)was applied to“Qihai”(CV6)and“Zhongji”(CV3)for CV group,“Mingmen”(GV4)and“Yaoshu”(GV2)for GV group,“non-acupoint 1”and“non-acupoint 3”on the left side for non-acupoint group,and manual acupuncture was applied to“Guanyuan”(CV4)for CV group,“Yaoyangguan”(GV3)for GV group,“non-acupoint 2”on the left side for non-acupoint group.The treatment was conducted for 20 min each time,once daily for 10 days.The writhing score was evaluated.The smooth myoelectric signals of rats’uterus in vivo were recorded by multi-channel physiological recorder.The uterine histopathological changes were observed by HE staining.The contents of prostaglandin F2α(PGF_(2α)),OT and calcium ion(Ca^(2+))in uterine tissue of rats were detected by ELISA.The protein and mRNA expression levels of smooth muscle 22-α(SM22-α),RhoA and ROCKⅡin uterine tissue were detected by Western blot and fluorescence quantitative PCR,respectively.Results Compared with the saline group,the writhing score of rats in the model group was increased(P<0.01),the amplitude voltage of uterine smooth muscle in vivo was elevated(P<0.01),the contents of PGF_(2α),OT and Ca^(2+),the protein and mRNA expression of SM22-α,RhoA and ROCKⅡin uterine tissue were all increased(P<0.01).Compared with the model and the non-acupoint groups,the writhing scores

关 键 词：原发性痛经 电针 RHO/RHO激酶信号通路 任脉 督脉 

分 类 号：R245.97[医药卫生—针灸推拿学]