疏肝调神针法对偏头痛大鼠受体活性修饰蛋白1、5-羟色胺1D受体表达的影响

Manual Acupuncture Stimulation Regulates Expression of Receptor Activity-modifying Protein 1 and 5-HT 1D Receptor Proteins and Genes in Migraine Rats

目的:探讨疏肝调神针法对偏头痛大鼠三叉神经脊束核、中脑受体活性修饰蛋白1(RAMP1)、5-羟色胺1D受体(5-HT1DR)表达的影响,探讨本法治疗偏头痛的镇痛机制。方法:雄性Wistar大鼠随机分为对照组、模型组、疏肝调神组、普通针刺组,每组10只。疏肝调神组取"百会""风池""内关""太冲"、普通针刺组取"百会""风池"进行针刺,每次30min,每日1次,共8d。针刺干预结束后,于大鼠颈后皮下注射硝酸甘油复制偏头痛模型。采用荧光定量PCR法、Western blot法检测大鼠三叉神经脊束核、中脑RAMP1、5-HT1D R的mRNA和蛋白表达情况。结果:与对照组比较,模型组三叉神经脊束核、中脑中RAMP1的mRNA、蛋白表达均显著升高(P<0.05),5-HT1DR的mRNA、蛋白表达均显著降低(P<0.05)。与模型组比较,疏肝调神组、普通针刺组三叉神经脊束核、中脑中RAMP1的mRNA、蛋白表达均明显降低(P<0.05),5-HT1DR的mRNA、蛋白表达均明显升高(P<0.05)。与普通针刺组比较,疏肝调神组三叉神经脊束核、中脑中RAMP1的mRNA、蛋白表达均明显降低(P<0.05),5-HT1DR的mRNA、蛋白表达均明显升高(P<0.05)。结论:疏肝调神针法对偏头痛的治疗作用可能与抑制中枢降钙素基因相关肽的表达、激活5-HT的表达有关。

Objective To observe the effect of liver-soothing and mental-activity-regulating（LSMAR）needling on the expression of receptor activity-modifying protein 1（RAMP 1,receptor of calcitonin gene-related peptide）,5-hydroxytryptamine1 Dreceptor（5-HT 1 DR）in the spinal trigeminal nucleus（STN）and midbrain in migraine rats,so as to explore its underlying mechanism in relieving migraine.Methods A total of 40 male Wistar rats were randomly divided into control,model,LSMAR and conventional needling groups（n=10 rats in each）.The migraine model was established by subcutaneous injection of nitroglycerin at the posterior neck.LSMAR was applied to＂Baihui＂（GV 20）,bilateral＂Fengchi＂（GB 20）,＂Neiguan＂（PC 6）and＂Taichong＂（LR 3）in the LSMAR group and conventional needling was applied to＂Baihui＂（GV 20）and bilateral＂Fengchi＂（GB 20）in the conventional acupuncture group for 30 min,once a day for 8 days before modeling.The expression levels of RAMP 1 and 5-HT 1 DR proteins and mRNAs in the STN and mesencephalon were detected by real-time fluorescent quantitative PCR and Western blot,separately.Results Compared with the control group,the expression levels of RAMP 1 protein and mRNA in STN and mesencephalon were significantly increased（P〈0.05）and those of 5-HT 1 DR protein and mRNA considerably decreased（P〈0.05）in the model group.After the acupuncture treatment,the increased levels of RAMP 1 protein and mRNA and the decreased levels of 5-HT 1 DR protein and mRNA in the STN and midbrain were obviously reversed in the LSMAR and conventional needling groups relevant to the model group（P〈0.05）.The effect of LSMAR needling was significantly superior to that of conventional needling in down-regulating the expression levels of RAMP 1 mRNA and protein in the STN and mesencephalon（P〈0.05）and in up-regulating the expression levels of 5-HT 1 DR mRNA and protein in the two brain regions（P〈0.05）.Conclusion Manual acupuncture stimulation of GV 20,GB 20,etc.can inhibit the expression of RAMP 1 protein and mRNA in the STN and midbrain,and up-regulate the expression of 5-HT 1 DR in the two brain regions of migraine rats,which may be related to its effect in relieving migraine.