愈风宁心滴丸预防性治疗血管痉挛性偏头痛的作用及机制研究  

Effect and mechanism of Yufeng Ningxin Dropping Pills on preventive treatment of vasospasm migraine

作　　者：蒋晓萌 杨代刚 高永耀 邹庭峰 施存元 江宏 段亚君 杨潇潇 JIANG Xiao-meng;YANG Dai-gang;GAO Yong-yao;ZOU Ting-feng;SHI Cun-yuan;JIANG Hong;DUAN Ya-jun;YANG Xiao-xiao(Zhejiang Jianfeng Pharmaceutical Co.,Ltd.,Jinhua 321000,China;Hefei University of Technology,Hefei 230001,China)

机构地区：[1]浙江尖峰药业有限公司,浙江金华321000 [2]合肥工业大学,安徽合肥230001

出　　处：《中草药》2021年第22期6881-6889,共9页Chinese Traditional and Herbal Drugs

基　　金：安徽省重点研究和开发计划项目(201904a07020007);中国博士后科学基金资助项目(2020M681914)。

摘　　要：目的观察愈风宁心滴丸对硝酸甘油诱导的偏头痛小鼠的影响,探索愈风宁心滴丸预防性治疗血管痉挛性偏头痛的作用及机制。方法采用脂多糖(lipopolysaccharide,LPS)刺激BV2神经小胶质细胞,以愈风宁心滴丸(10、50、100μmol/L)及葛根素(50μmol/L)进行干预,收集细胞。将野生型C57BL/6J小鼠随机分成照组、模型组以及愈风宁心滴丸低、中、高剂量(200、360、600 mg/kg)组和葛根素组(37.8 mg/kg),给予药物进行干预,于第3、5、7、9、11天ip硝酸甘油溶液(10 mg/kg)建立小鼠偏头痛模型,第3天开始进行痛觉敏感(机械压痛和热敏耐受)行为学实验;实验结束后,取小鼠血清和脑组织。检测各组BV2细胞活性氧含量;采用Western blotting法检测各组细胞超氧化物歧化酶1(superoxide dismutase 1,SOD1)、SOD2蛋白表达以及各组小鼠三叉神经脊束尾核(trigeminal nucleus caudalis,TNC)区降钙素基因相关肽(calcitonin gene-related peptide,CGRP)、c-Fos、磷酸化细胞外信号调节激酶(phosphorylated extracellular regulated protein kinases,p-ERK)及促炎细胞因子白细胞介素-6(interleukin-6,IL-6)蛋白表达;采用qRT-PCR法检测各组细胞SOD1、SOD2、IL-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)mRNA表达以及小鼠TNC区IL-1β、TNF-αmRNA表达;采用免疫荧光检测各组小鼠TNC区CGRP、c-Fos表达;采用生化分析仪检测各组小鼠血清中丙氨酸转氨酶(alanine aminotransferase,ALT)、天冬氨酸转氨酶(aspartate aminotransferase,AST)和碱性磷酸酶(alkalinephosphatase,ALP)活性。结果愈风宁心滴丸显著上调BV2细胞抗氧化酶SOD1、SOD2mRNA和蛋白表达水平(P<0.05、0.01、0.001),降低活性氧水平(P<0.01、0.001),抑制炎症因子IL-1β和TNF-αmRNA表达(P<0.001)。与对照组相比,模型组小鼠基础痛阈值显著降低(P<0.05、0.001),TNC区中CGRP、c-Fos、p-ERK蛋白表达水平显著升高(P<0.01、0.001),TNC区IL-1β、TNF-αmRNA和IL-6蛋白表达水平显著升高(PObjectiveTo observe the effect of Yufeng Ningxin Dropping Pills(愈风宁心滴丸)on nitroglycerin-induced migraine mice,and explore the effect and mechanism of Yufeng Ningxin Dropping Pills on preventive treatment of vasospasm migraine.Methods Lipopolysaccharide(LPS)was used to stimulate BV2 neuroglial cells,Yufeng Ningxin Dripping Pills(10,50,100μmol/L)and puerarin(50μmol/L)were used for intervention,and cells were collected.Wild-type C57BL/6J mice were randomly divided into control group,model group,low-,medium-,and high-dose(200,360,600 mg/kg)Yufeng Ningxin Dripping Pill group and puerarin(37.8 mg/kg)group,drugs were given for intervention,migraine model of mice was established with ip nitroglycerin solution(10 mg/kg)on 3rd,5th,7th,9th and 11th day,hyperalgesia tolerance behavioral experiment(mechanical tenderness and heat sensitivity)were started on 3rd day;After experiment,the serum and brain tissue of mice were taken.The content of reactive oxygen species in BV2 cells of each group was detected;Western blotting was used to detect the expressions of superoxide dismutase 1(SOD1)and SOD2 protein in BV2 cells and calcitonin gene-related peptide(CGRP),c-Fos,phosphorylated extracellular regulated protein kinases(p-ERK)and pro-inflammatory cytokine interleukin-6(IL-6)protein expressions in trigeminal nucleus caudalis(TNC)region in mice of each group;qRT-PCR method was used to detect SOD1,SOD2,IL-1β,tumor necrosis factor-α(TNF-α)mRNA expressions in BV2 cells and IL-1βand TNF-αmRNA expressions in TNC of mice;Immunofluorescence was used to detect the expressions of CGRP and c-Fos in TNC area of micein each group;Automatic biochemical analyzerwas used to detect the alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)activities in serum of mice.Results Yufeng Ningxin Dripping Pills significantly increased antioxidant enzyme SOD1and SOD2mRNA and protein expression levels in BV2 cells(P<0.05,0.01,0.001),reduced reactive oxygen species levels(P<0.01,0.001),inhibited the infla

关 键 词：愈风宁心滴丸 偏头痛 神经小胶质细胞 血管扩张 神经炎症 

分 类 号：R285.5[医药卫生—中药学]