瞬时受体电位通道M2在动脉粥样硬化小鼠血管高反应性中的作用  

Function of transient receptor potential melastatin 2 in vascular hyper-reactivity during the development of mouse atherosclerosis

作　　者：戴贻权[1] 颜晓晓 林奕辰 陈宏宇[1] 刘晓如[3] DAI Yi-quan;YAN Xiao-xiao;LIN Yi-chen;CHEN Hong yu;LIU Xiao ru(Department of Vascular Surgery,The First Affiliated Hospital of FujianMedical University,Fuzhou Fujian 350005,China)

机构地区：[1]福建医科大学附属第一医院血管外科,福建福州350005 [2]福建省儿童医院病理科 [3]福建医科大学基础医学院

出　　处：《中华高血压杂志》2020年第7期653-660,共8页Chinese Journal of Hypertension

基　　金：福建省自然科学基金(2017J01281,2019J01296)。

摘　　要：目的探究瞬时受体电位通道M2(TRPM2)在动脉粥样硬化(AS)小鼠主动脉5-羟色胺高反应性中的作用。方法将40只小鼠随机分为C57BL/6J普通饲料组(C57+nd)、C57BL/6J高脂饲料组(C57+hfd)、载脂蛋白E基因敲除(APOE-/-)普通饲料组(APOE-/-+nd)和APOE-/-高脂饲料组(APOE-/-+hfd),喂养16周。比较4组小鼠主动脉TRPM2基因表达变化,运用血管环张力检测技术测定小鼠主动脉对5-羟色胺的反应性变化,观察TRPM2抑制剂处理对5-羟色胺反应性的作用。结果APOE-/-+hfd组小鼠体质量、血糖和血脂显著增高,主动脉斑块形成明显,模型制备成功。APOE-/-+hfd组小鼠主动脉TRPM2 mRNA(3.20±0.97)与蛋白(2.82±0.35)相对表达水平高于其他3组(mRNA:C57+nd组0.92±0.42、C57+hfd组0.74±0.37、APOE-/-+nd组0.87±0.42;蛋白:C57+nd组1.39±0.23、C57+hfd组1.35±0.34、APOE-/-+nd组1.22±0.23);APOE-/-+hfd组主动脉对5-羟色胺反应性增强,半最大效应浓度(EC50)减小,且可被TRPM2抑制剂N-苯基邻氨基苯甲酸(ACA,1μmol/L)和2-氨基乙酯二苯基硼酸(2-APB,10μmol/L)显著抑制[ACA抑制率:C57+nd组(16.62±1.28)%、C57+hfd组(19.20±5.55)%、APOE-/-+nd组(14.72±2.30)%、APOE-/-+hfd组(38.15±1.13)%,F=12.58,P<0.01;2-APB抑制率:C57+nd组(27.68±5.24)%、C57+hfd组(28.75±3.67)%、APOE-/-+nd组(50.40±7.74)%、APOE-/-+hfd组(62.81±5.99)%,F=98.83,P<0.01]。结论APOE基因敲除和高脂喂食处理可能通过上调TRPM2基因表达,增强TRPM2在5-羟色胺高反应性中的作用,促进小鼠AS的产生。Objective To investigate the function of transient receptor potential melastatin 2(TRPM2)in 5-hydroxytryptamine(5-HT)-induced hyper-reactivity of aorta contraction during the development of mouse atherosclerosis(AS).Methods Forty mice were randomly divided into 4 groups:C57 BL/6 J on normal diet(C57+nd),C57 BL/6 J on high-fat diet(C57+hfd),apolipoprotein E gene knockout mice(APOE-/-)on normal diet(APOE-/-+nd)and APOE-/-on high-fat diet(APOE-/-+hfd).They were fed with normal diet or high-fat diet respectively for 16 weeks.TRPM2 expression in aortas was detected with real-time reverse transcription polymerase chain reaction(RT-PCR)and Western blot.The response of mouse aorta to 5-HT was measured by isometric contraction,and the effect of TRPM2 inhibitor treatment on vascular response to 5-HT was detected.Results In APOE-/-+hfd group,the body weight,blood glucose and blood lipid increased,and the development of aortic plaque was obvious,which suggested the atherosclerosis model was successfully established.Compared with the other 3 groups,the expressions of TRPM2 mRNA(3.20±0.97)and protein(2.82±0.35)in aortas of APOE-/-+hfd group were critically increased(mRNA:C57+nd group 0.92±0.42,C57+hfd group 0.74±0.37,APOE-/-+nd group 0.87±0.42;protein:C57+nd group 1.39±0.23,C57+hfd group 1.35±0.34,APOE-/-+nd group 1.22±0.23).The reactivity to 5-HT in aorta of APOE-/-+hfd group was enhanced and the concentration for 50%of maximum effect(EC50)was reduced.The reactivity to 5-HT could be significantly inhibited by the TRPM2 inhibitors,N-p-amylcinnamoyl anthranilic acid(ACA,1μmol/L)and 2-aminoethyl diphenylbrinate(2-APB,10μmol/L)[inhibition of ACA:C57+nd group(16.62±1.28)%,C57+hfd group(19.20±5.55)%,APOE-/-+nd group(14.72±2.30)%,APOE-/-+hfd group(38.15±1.13)%,F=12.58,P<0.01;inhibition of 2-APB:C57+nd group(27.68±5.24)%,C57+hfd group(28.75±3.67)%,APOE-/-+nd group(50.40±7.74)%,APOE-/-+hfd group(62.81±5.99)%,F=98.83,P<0.01].Conclusion ApoE gene knockout and high-fat diet may promote the development of AS in mice by u

关 键 词：动脉粥样硬化 瞬时受体电位通道M2 5-羟色胺 主动脉平滑肌细胞 

分 类 号：R543.5[医药卫生—心血管疾病]