川芎-天麻药对对偏头痛模型大鼠眶周机械痛阈及三叉神经节AMPK/TRPA1通路的影响
The Effect of Chuanxiong(Rhizoma Chuanxiong)& Tianma(Rhizoma Gastrodiae)Herbal Pair on the Periorbital Mechanical Pain Threshold and AMPK/TRPA1 Pathway in the Trigeminal Ganglion of Migraine Model Rats
左笑 1梁丽琦 1唐岚琦 1蒙睿娜 1房冰莹 1王海芳 1崔海燕 2刘晓华1
作者信息
- 1. 陕西中医药大学,陕西省咸阳市秦都区世纪大道中段1号,712046
- 2. 西藏民族大学
- 折叠
摘要
目的 基于腺苷酸活化蛋白激酶(AMPK)/瞬时受体电位A1通道(TRPA1)通路探究川芎-天麻药对治疗偏头痛的可能作用机制.方法 48只健康雄性SD大鼠随机分为对照组、模型组、川芎-天麻药对组,每组16只.对照组和模型组给予10 ml/kg生理盐水灌胃,川芎-天麻药对组给予0.675 g/kg川芎-天麻灌胃,均每天1次,连续8天.末次给药前造模,对照组皮下注射生理盐水10ml/kg,模型组和川芎-天麻药对组皮下注射硝酸甘油10 ml/kg制备偏头痛模型大鼠.运用Von Frey纤毛测定大鼠眶周机械痛阈值;采用酶联免疫吸附测定法(ELISA)测定大鼠血清、脑脊液中降钙素基因相关肽(CGRP)水平;一氧化氮(NO)试剂盒测定血清、脑脊液中NO水平;RT-PCR法检测大鼠三叉神经节中即刻早期基因(c-Fos)、CGRP、瞬时受体电位V1通道(TRPV1)、腺苷酸活化蛋白激酶A(PRKAA)及TRPA1 mRNA表达水平;免疫荧光法检测三叉神经颈复合体(TCC)中c-Fos阳性细胞数和三叉神经节磷酸化腺苷酸活化蛋白激酶(pAMPK)、TRPA1蛋白表达水平.结果 与对照组比较,模型组大鼠机械刺激阈值、pAMPK蛋白表达降低,血清中CGRP、NO水平,三叉神经节c-Fos、CGRP、TRPV1、TRPA1 mRNA表达水平,TRPA1蛋白表达和TCC中c-Fos阳性细胞数均显著升高(P＜0.05).与模型组比较,川芎-天麻药对组大鼠机械刺激阈值、pAMPK蛋白表达显著上升,血清中CGRP、NO水平,三叉神经节中c-Fos、CGRP、TRPV1、TRPA1 mRNA表达水平,TRPA1蛋白表达和TCC中c-Fos阳性细胞数明显下降(P＜0.05).结论 川芎-天麻药对可能通过调控三叉神经节中AMPK/TRPA1通路,升高机械痛阈,从而改善偏头痛症状.
Abstract
Objective To explore the possible mechanism of Chuanxiong(Rhizoma Chuanxiong)& Tianma(Rhizoma Gastrodiae)herbal pair in treating migraines based on AMP-activated protein kinase(AMPK)/transient receptor potential Al channel(TRPA1)pathway.Methods Forty-eight healthy male SD rats were randomly divided into control group,model group,and Chuanxiong Tianma medication group,with 16 rats in each group.The control group and model group were given 10 ml/kg of normal saline by gavage,while the Chuanxiong Tianma medication group was given 0.675 g/kg of Chuanxiong Tianma herbal pair by gavage,once daily for 8 consecutive days in both groups.Migraime model was performed before the last administration,with subcutaneous injection of 10 ml/kg of normal saline in the control group,and subcutaneous injection of 10 ml/kg of nitroglycerin in the model group and Chuanxiong Tianma medication group.The Von Frey filament was used to measure the periorbital mechanical pain threshold of rats.The enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of calcitonin gene-related peptide(CGRP)in rat serum and cerebrospinal fluid.The nitric oxide(NO)assay kit was used to deter-mine the NO level in serum and cerebrospinal fluid.RT-PCR was usedto detect the mRNA expression levels of immediate-early genes in the trigeminal ganglion of rats(c-Fos),CGRP,transient receptor potential V1 channel(TRPV1),AMPK alpha subunit(PRKAA),and TRPA1.Immunofluorescence was used to detect the number of c-Fos-positive cells in the trigeminal cervical complex(TCC)and the protein expression levels of phosphorylated AMPK(pAMPK)and TRPA1 in the trigeminal ganglion.Results Compared to those in the control group,the mechanical stimulation threshold and pAMPK protein expression in the model group decreased,while the levels of CGRP and NO in serum,c-Fos,CGRP,TRPV1 and TRPA1 mRNA levels in the trigeminal ganglion,TRPA1 protein expression,and the number of c-Fos-positive cells in the TCC significantly increased(P＜0.05).Compared to those in the model group,the mechanical stimulation threshold and pAMPK protein expression in the Chuanxiong Tianma medication group signifi-cantly increased,while the levels of CGRP and NO in serum,c-Fos,CGRP,TRPV1 and TRPA1 mRNA levels in the tri-geminal ganglion,TRPA1 protein expression,and the number of c-Fos-positive cells in the TCC significantly decreased(P＜0.05).Conclusion Chuanxiong Tianma herbal pair may improve migraine symptoms by regulating the AMPK/TRPA1 pathway in the trigeminal ganglion and increasing the mechanical pain threshold.
关键词
偏头痛/川芎/天麻/三叉神经节/痛阈/腺苷酸活化蛋白激酶/瞬时受体电位A1通道Key words
migraine/Chuanxiong(Rhizoma Chuanxiong)/Tianma(Rhizoma Gastrodiae)/trigeminal ganglion/pain threshold/adenosine AMP-activated protein kinase/transient receptor potential A1 channel引用本文复制引用
基金项目
国家自然科学基金(8200422)
陕西省教育厅重点科学研究计划项目(23JS005)
陕西中医药大学大学生创新创业训练计划项目(S202210716057)
西藏自治区自然科学基金(ZRKX2020000075)
陕西省自然科学基础研究计划项目(2022JQ-870)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据