基于热敏通道TRPV1探讨中药甘遂止痛机理  

Gansui(Euphorbia kansui) Extracts Activate the Transient Receptor Potential Vanilloid 1

作　　者：韩雪珍 佟海英[1] 高蔚[1] 马少丹[2] 郭蓉[1] 殷茵 安致君 杨桢[1] 刘珍洪[1] HAN Xuezhen;TONG Haiying;GAO Wei;MA Shaodan;GUO Rong;YIN Yin;AN Zhijun;YANG Zhen;LIU Zhenhong(Beijing University of Chinese Medicine,Beijing 100029,China;Fujian University of Chinese Medicine,Fuzhou 100029,Fujian,China)

机构地区：[1]北京中医药大学,北京100029 [2]福建中医药大学,福建福州350122

出　　处：《中华中医药学刊》2020年第7期126-130,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81573847);北京中医药薪火传承“3+3”工程李庆业名老中医工作站项目(2009-JC-31);北京中医药科技发展资金项目(JJ2015-56)。

摘　　要：目的探讨中药甘遂对热敏通道瞬时感受器电位香草素受体1(TRPV1)的影响。方法以电生理全细胞膜片钳技术检测不同浓度中药甘遂的75%乙醇提取物对瞬染人源TRPV1的HEK293细胞(hTRPV1/HEK293细胞)诱导产生的跨膜电流,运用TRPV1特异性拮抗剂辣椒平,观察其是否可以抑制中药甘遂诱导产生的跨膜电流,使用C57BL/6小鼠30只,观察在(0±2)℃冷板和(55±1)℃热板中小鼠冷痛、热痛行为潜伏期的变化。结果全细胞膜片钳实验显示在hTRPV1/HEK293细胞上甘遂75%乙醇提取物可以激活TRPV1通道产生明显的跨膜电流,该跨膜电流与TRPV1激动剂辣椒素诱发的电流在电流密度和电流-电压关系各方面均类似;量效实验显示10 mg·mL^-1甘遂醇提物能够激活hTRPV1/HEK293细胞诱导其产生明显的跨膜电流(P<0.001)。10 mg·mL^-1甘遂醇提物产生的跨膜电流>3 mg·mL^-1,差异具有极显著统计学意义(P<0.001)。TRPV1特异性拮抗剂辣椒平可以完全抑制10 mg·mL^-1甘遂醇提物诱导产生的跨膜电流;小鼠冷板和热板实验中,甘遂延长小鼠的冷痛和热痛行为潜伏期与剂量呈现一定的量效关系。与空白组小鼠对比,甘遂中、高剂量组和布洛芬阳性对照组小鼠冷痛行为潜伏期明显延长,差异具有统计学意义(P<0.05,P<0.001,P<0.05)。甘遂中、高剂量组和布洛芬阳性对照组的热痛行为潜伏期明显延长(P<0.05,P<0.01,P<0.05)。结论中药甘遂的75%乙醇提物中含有TRPV1的激动剂,甘遂所具备的止痛和抗炎功效可能是热敏通道TRPV1活化后产生的系列效应。Objective To investigate the effect of ethanol extracts of Gansui(Euphorbia kansui)on thermosensitive channeltransient receptor potential vanilloid 1(TRPV1).Methods Whole cell voltage clamp technique was used to record transmembrane currents induced by the different concentrations of 75%ethanol extracts of Gansui(Euphorbia kansui)in HEK293 cells which expressed human TRPV1(hTRPV1/HEK293).By using TRPV1 specific antagonist capsaicin,we observed whether it could inhibit the transmembrane current induced by Gansui(Euphorbia kansui).Thirty C57 BL/6 mice were used to observe the changes of behavioral latency of cold pain and heat pain in a cold plate device at(0±2)℃and a hot plate device at(55±1)℃.Results Whole cell patch clamp experiment showed that 75%ethanol extract of Gansui(Euphorbia kansui)on hTRPV1/HEK293 cells could activate TRPV1 channel to generate obvious transmembrane current,compared with that by the known TRPV1 agonistcapaicinin all aspects of current density and current-voltage relationship.Dose-response experiments showed that 10 mg·mL^-1 ethanol extract of Gansui(Euphorbia kansui)could activate hTRPV1/HEK293 cells to induce significant transmembrane current(P<0.001).The transmembrane current generated by 10 mg·mL^-1 ethanol extract of Gansui(Euphorbia kansui)was more than that of 3 mg·mL^-1ethanol extract(P<0.001).The TRPV1 specific antagonist capsaicin could completely inhibit the transmembrane current induced by 10 mg·mL^-1 ethanol extract of Gansui(Euphorbia kansui)in the experiment of cold plate and hot plate in mice.There was a dose-effect relationship between the latency of cold pain behavior and hot pain behavior in mice prolonged by Gansui(Euphorbia kansui).Compared with the control group,the latency of cold pain behavior of Gansui(Euphorbia kansui)was significantly longer in medium and high dose groups and ibuprofen positive control groups(P<0.05,P<0.001,P<0.05).The latency of heat pain behavior in medium and high dose groups of Gansui(Euphorbia kansui)and positive control group of

关 键 词：甘遂 热敏通道TRPV1 激动剂 止痛 

分 类 号：R285.5[医药卫生—中药学]