电针对偏头痛大鼠三叉神经节miR-34a-5p/SIRT1通路的影响  

Effects of electroacupuncture on miR-34a-5p/SIRT1 signaling in the trigeminal ganglion of rats with migraine

作　　者：张慧 何胜东 纵单单[3] 张雪梅 罗婧 郑佳坤 ZHANG Hui;HE Sheng-dong;ZONG Dan-dan;ZHANG Xue-mei;LUO Jing;ZHENG Jia-kun(College of Integrated Traditional Chinese and Western Medicine,Southwest Medical University,Luzhou 646000,Sichuan Province,China;The Second Affiliated Hospital,Guilin Medical University,Guilin 541100,Guangxi Province;The Second Xiangya Hospital,Central South University,Changsha 410038;Graduate School,Beijing University of Chinese Medicine,Beijing 100078)

机构地区：[1]西南医科大学中西医结合学院,四川泸州646000 [2]桂林医学院第二附属医院,广西桂林541100 [3]中南大学湘雅二医院,长沙410038 [4]北京中医药大学研究生院,北京100078

出　　处：《针刺研究》2020年第11期868-874,共7页Acupuncture Research

基　　金：国家自然科学基金青年基金项目(No.81704173);四川省教育厅重点项目(No.17ZA0662)。

摘　　要：目的:观察电针对偏头痛大鼠三叉神经节miR-34a-5p、沉默信息调节因子1(SIRT1)及核因子-κB亚基p65(NF-κB p65)表达的影响,探讨电针预防偏头痛的作用机制。方法:雄性SD大鼠随机分为正常组、假手术组、模型组、电针组及电针+EX527(SIRT1抑制剂)组,每组10只。电刺激三叉神经节建立偏头痛大鼠模型。电针组、电针+EX527组大鼠建模前连续5 d分别给予电针"外关""风池"干预,每次20 min,1次/d,电针+EX527组同时每天腹腔注射EX527(5 mg/kg)。建模后第1天处死大鼠,酶联免疫吸附法检测大鼠血清前列腺素E2(PGE2)和降钙素基因相关肽(CGRP)的浓度,实时荧光定量PCR法检测三叉神经节miR-34a-5p及SIRT1、白细胞介素-1β(IL-1β)的mRNA水平,Western blot法检测三叉神经节SIRT1、IL-1β、NF-κB p65、乙酰化NF-κB p65(NF-κB Ac-p65)和环氧化酶2(COX2)蛋白表达。结果:与正常组和假手术组比较,模型组大鼠血清PGE2和CGRP浓度升高(P<0.05),三叉神经节miR-34a-5p表达增加(P<0.05)、SIRT1 mRNA和蛋白表达减少(P<0.05)、IL-1βmRNA和蛋白表达增加(P<0.05),NF-κB p65、NF-κB Ac-p65和COX2蛋白表达增加(P<0.05);与模型组比较,电针组血清PGE2和CGRP浓度降低(P<0.05),三叉神经节miR-34a-5p表达减少(P<0.05)、SIRT1 mRNA和蛋白表达增加(P<0.05)、IL-1βmRNA和蛋白表达减少(P<0.05)、NF-κB p65、NF-κB Ac-p65和COX2蛋白表达减少(P<0.05);与电针组比较,电针+EX527组血清PGE2和CGRP浓度升高(P<0.05),三叉神经节IL-1βmRNA和蛋白表达增加(P<0.05),SIRT1蛋白表达减少(P<0.05),NF-κB p65、NF-κB Ac-p65和COX2蛋白表达增加(P<0.05)。结论:电针可抑制偏头痛大鼠三叉神经节miR-34a-5p表达,增加SIRT1表达,下调IL-1β/COX2炎性反应信号,减少PGE2合成,进而减少三叉神经末梢释放CGRP,这可能是电针预防偏头痛的机制之一。Objective To investigate the effect of electroacupuncture(EA)on miR-34 a-5 p,silent mating type information regulation 2 homolog-1(SIRT1)and nuclear factor-κB subunit p65(NF-κB p65)in the trigeminal ganglion of rats with migraine,so as to explore the mechanisms of EA underlying prevention of migraine.Methods Male SD rats were randomly divi-ded into normal,sham operation,model,EA,and EA plus EX527(a SIRT1 inhibitor)groups,with 10 rats in each group.The rat model of migraine was established by electrical stimulation of the trigeminal ganglion.Before modeling,EA was applied at"Waiguan"(TE5)and"Fengchi"(GB20)for 20 min each time,once a day for 5 consecutive days,and intraperitoneal injection of EX527(5 mg/kg)every day simultaneously.Serum prostaglandin E2(PGE2)and calcitonin gene-related peptide(CGRP)concentrations were measured by enzyme-linked immunosorbent assay.The levels of miR-34 a-5 p,SIRT1 and interleukin-1β(IL-1β)mRNA,and protein expression of SIRT1,IL-1β,NF-κB p65,NF-κB Ac-p65 and cyclooxygenase-2(COX2)in trigeminal ganglia were detected by real-time quantitative PCR and Western blot,separately.Results The serum concentrations of PGE2 and CGRP,the expression of miR-34 a-5 p,IL-1βmRNA and protein,NF-κB p65,NF-κB Ac-p65 and COX2 protein expression in the trigeminal ganglion were remarkably increased(P<0.05),while the SIRT1 mRNA and protein decreased(P<0.05)in the model group in contrast to the normal group.Following EA intervention,the serum PGE2 and CGRP concentrations,miR-34 a-5 p expression,IL-1βmRNA and protein,NF-κB p65,NF-κB Ac-p65 and COX2 protein expression were significantly down-regulated(P<0.05),and SIRT1 mRNA and protein significantly up-regulated(P<0.05).Compared with the EA group,the serum concentrations of PGE2 and CGRP,IL-1βmRNA and protein,NF-κB p65,NF-κB Ac-p65 and COX2 protein expressions increased(P<0.05),and SIRT1 protein decreased(P<0.05)in the EA plus EX527 group.Conclusion In migraine rats,EA can inhibit miR-34 a-5 p expression in the trigeminal ganglion,increase SIRT1

关 键 词：偏头痛 电针 三叉神经节 miR-34a-5p 沉默信息调节因子1 核因子-κB亚基p65 

分 类 号：R245.9+7[医药卫生—针灸推拿学]