推拿对神经病理性疼痛大鼠miR-21-5P、ERK、STAT3的影响  

Effects of Massage on miR-21-5P, ERK and STAT3 in Rats with Neuropathic Pain

作　　者：吴娉婷 肖易[2] 梁英业 谢柳红 丁瑶 卢栋明 陈晓红 唐宏亮[3] Wu Pingting;Xiao Yi;Liang Yingye;Xie Liuhong;Ding Yao;Lu Dongming;Chen Xiaohong;Tang Hongliang(Guangxi University of Chinese Medicine,Nanning 530000,China;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530000,China;Fangchenggang Hospital affiliated to Guangxi University of Chinese Medicine,Fangchenggang 538000,China)

机构地区：[1]广西中医药大学,南宁530000 [2]广西中医药大学第一附属医院,南宁530000 [3]广西中医药大学附属防城港医院,防城港538000

出　　处：《世界科学技术-中医药现代化》2022年第11期4393-4401,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会青年科学基金项目(8180151635):从LncBANCR介导ERK/mTOR调控自噬探讨推拿对SNL大鼠的镇痛效应机制,负责人:卢栋明;广西壮族自治区自然科学基金委员会面上项目(2018JJA140664):从miRNA-21的表达调控作用探讨推拿治疗慢性病理性疼痛的效应机制研究,负责人:肖易;广西壮族自治区研究生教育创新计划区级项目(YCSW2021229):从LNCRNAH19调控miR-342-3p/IER3探讨枢经推拿对NPP的镇痛机制研究,负责人:吴娉婷。

摘　　要：目的 观察在神经病理性疼痛大鼠中miR-21-5P、细胞外调节蛋白激酶(Extracellular signal-regulated kinases,ERK)和信号转导和转录活化因子3(Signal transducer and activator of transcription3,STAT3)的表达变化,探究推拿镇痛作用的机制。方法 40只SD雄性大鼠,按照随机分类的方法将其分为正常组、模型组、假手术组、推拿组和假推拿组5个组,每组8只。不给予正常组任何干预措施;模型组、推拿组及假推组建立L5脊神经结扎模型(SNL);只暴露假手术组的脊神经2-3 min;在造模后第1天使用按摩推拿手法模拟仪对推拿组的“环跳”“、阳陵泉”和“悬钟”穴进行推拿治疗,每天1次,每次治疗18 min(双侧),连续干预7天;假推拿组的大鼠则每天给予双后肢18 min的轻轻抚摸,连续干预7天。各组大鼠的机械痛阈值(Paw withdrawl mechanical threshold,PWMT)和热痛阈值(Hindpaw withdrawal thermal latency,PWTL)在造模前1天、造模后第3天和第7天进行测定。使用实时荧光定量PCR(Quantitative Real-time PCR,q-PCR)检测造模后第7天各组大鼠中miR-21-5P的表达水平;使用蛋白免疫印迹法(Western blot)测定各组大鼠中ERK、STAT3的蛋白表达水平,酶联免疫吸附法(Enzyme linked immunosorbent assay,ELISA)检测各组大鼠中白细胞介素-1β(Interleukin-1β,IL-1β)的表达水平。结果 术后第3天和第7天,模型组大鼠的机械阈值与热痛阈值比正常组及假手术组的阈值下降(P<0.05),推拿组大鼠的机械阈值与热痛阈值比模型组、假推拿组的阈值升高(P<0.05)。术后第7天,q-PCR检测结果显示,模型组大鼠的miR-21-5P水平较正常组及假手术组的miR-21-5P水平显著增加(P<0.01),推拿组大鼠的miR-21-5P水平较模型组与假推拿组的miR-21-5P水平显著下降(P<0.01);Western blot检测结果显示,与正常组及假手术组比较,模型组大鼠的ERK与STAT3水平显著增加(P<0.01),与模型组、假推拿组比较,推拿组能显著下调大鼠ERK(P<0Objective To observe the expression changes of Mir-21-5p, ERK and STAT3 in neuropathic pain rats, and explore the analgesic mechanism of massage. Methods Forty male SD rats were randomly divided into 5 groups,including normal group, model group, sham operation group, massage group and sham massage group, with 8 rats in each group. The normal group was not given any intervention;L5 spinal nerve ligation model(SNL) was established in model group, massage group and false push group. Only spinal nerves of the sham group were exposed for 2-3 minutes;On the first day after modeling, massage manipulation simulator was used to massage the "Huantiao", "Yangling quan" and "Xuanzhong" points of the massage group, once a day, each treatment 18 min(bilateral), continuous intervention for 7 days;The rats in the sham massage group were gently stroked with both hind limbs for 18min every day for 7 consecutive days. PWMT and PWTL of rats in each group were determined 1 day before modeling, 3 and 7 days after modeling. The expression level of miR-21-5P in each group was detected by q-PCR on day 7 after modeling. The protein expression levels of ERK and STAT3 were determined by Western blot. The expression level of IL-1β in each group was detected by ELISA. Results On the 3rd and 7th day after operation, the mechanical threshold and thermal pain threshold of rats in the model group were lower than those in the normal group and the sham group(P<0.05), and the mechanical threshold and thermal pain threshold of rats in the massage group were higher than those in the model group and the sham group(P<0.05). On the 7th day after surgery, q-PCR results showed that miR-21-5p level in model group was significantly increased compared with normal group and sham group(P<0.01). The miR-21-5p level in massage group was significantly decreased compared with that in model group and sham massage group(P<0.01). Western blot test results showed that compared with normal group and sham operation group, ERK and STAT3 levels in model group were signifi

关 键 词：推拿 神经病理性疼痛 miR-21-5P ERK STAT3 IL-1Β 镇痛 

分 类 号：R244.1[医药卫生—针灸推拿学]