P2X3受体介导电针调节肠易激综合征内脏痛大鼠结肠5-HT、SERT的作用研究  

Regulation of 5-HT and SERT in colon of rats with IBS by P2X3 receptor-mediated electroacupuncture

作　　者：周云 张方 闫翠娜 李琪[1] 乔宇 黄榕 翁志军[1,2] 杨玲 刘慧荣[1,2] ZHOU Yun;ZHANG Fang;YAN Cuina;LI Qi;QIAO Yu;HUANG Rong;WENG Zhijun;YANG Ling;LIU Huirong(Key Laboratory of Acupuncture Immune Effects,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200030,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院针灸免疫效应重点研究室,200030 [2]上海市针灸经络研究所

出　　处：《环球中医药》2023年第9期1715-1722,共8页Global Traditional Chinese Medicine

基　　金：上海市青年科技英才扬帆计划项目(18YF1421600);国家自然科学基金青年项目(81904301);上海市自然科学基金(22ZR1458600);上海市针灸临床医学研究中心(20MC1920500)。

摘　　要：目的探讨嘌呤能配体门控离子通路3(purinergicligand-gatedionchannel 3,P2X3)受体介导电针对肠易激综合征(irritable bowel syndrome,IBS)内脏痛大鼠结肠5-羟色胺(5-hydroxytryptamine,5-HT)、5-HT再摄取转运蛋白(serotonnin transporter,SERT)的调节作用。方法选取SPF级SD雄性大鼠,随机分为正常组、模型组、电针组、P2X3受体激动剂组、P2X3受体激动剂+电针组,通过三硝基苯磺酸(trinitrobrnzen sulfonic acid,TNBS)灌肠液灌肠制备IBS内脏痛大鼠模型。模型制备3周后运用电针、髓鞘内注射、电针和髓鞘内注射相结合等方法进行干预。采用结肠组织病理染色、腹部回撤反射(abdominal withdrawal reflex,AWR)评分观察IBS内脏痛模型是否制作成功,运用AWR评分、酶联免疫吸附法、蛋白质印迹法、实时荧光定量PCR等方法检测IBS大鼠内脏痛敏的变化及IBS大鼠结肠5-HT、SERT的表达变化。结果(1)电针能降低IBS大鼠在40、80 mmHg压力结直肠扩张(colorectal distention,CRD)刺激下的AWR评分(P<0.05,P<0.01);(2)电针能够下调IBS大鼠结肠5-HT的表达(P<0.01),上调结肠SERT蛋白和mRNA的表达(P<0.01);(3)P2X3受体被激活后,电针能够下调IBS大鼠结肠5-HT的表达(P<0.05),上调结肠SERT蛋白和mRNA的表达(P>0.05)。结论P2X3受体介导电针调节IBS内脏痛大鼠结肠中5-HT、SERT蛋白及mRNA的异常表达,可能是电针缓解IBS大鼠内脏痛外周敏化的机制之一。Objective To investigate the regulation of 5-HT and SERT in colon of rats with irritable bowel syndrome(IBS)by P2X3 receptor-mediated electroacupuncture.Methods SPF male SD rats were randomly divided into normal group,model group,electroacupuncture group,P2X3 receptor agonist group,and P2X3 receptor agonist+electroacupuncture group.The visceral pain model of IBS was prepared by intestinal enema with trinitrobrnzen sulfonic acid(TNBS).Three weeks after model preparation,electroacupuncture,intramedullary injection,electroacupuncture and intramedullary injection were used for intervention.Hematoxylin-eosin staining of colon and abdominal withdrawal reflex(AWR)scores were used to observe the success of visceral pain model of IBS.AWR scores,ELISA,Western Blot,real-time PCR and other methods were used to detect the changes of visceral pain sensitivity and the expression of 5-HT and SERT in the colon of rats with IBS.Results(1)Electroacupuncture can reduce the AWR scores of rats with IBS under CRD stimulation at 40 mmHg and 80 mmHg pressure(P<0.05,P<0.01).(2)Electroacupuncture can down-regulate the expression of 5-HT(P<0.01)and up-regulate the expression of SERT protein and mRNA in the colon of rats with IBS(P<0.05).(3)After activation of P2X3 receptor,electroacupuncture can down-regulate the expression of 5-HT(P<0.05)and up-regulate the expression of SERT protein and mRNA in the colon of rats with IBS(P>0.05).Conclusion Electroacupuncture regulates the abnormal expression of 5-HT and SERT protein and mRNA in the colon of IBS visceral pain rats by P2X3 receptor mediated,which may be one of the mechanisms of electroacupuncture to relieve peripheral sensitization of visceral pain in rats with IBS.

关 键 词：肠易激综合征 内脏痛 电针 嘌呤能配体门控离子通路3受体 5-羟色胺 5-羟色胺再摄取转运蛋白 

分 类 号：R245[医药卫生—针灸推拿学]