电针对阿片类药物耐受大鼠海马G蛋白偶联受体及cAMP、CREB表达的影响  

Effects of Electroacupuncture on Hippocampal G Protein-coupled Receptors,cAMP and CREB Expression in Opioid-resistant Rats

作　　者：刘春[1] 于波[1] 王铁东[1] LIU Chun;YU Bo;WANG Tie-dong(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning 110032)

机构地区：[1]辽宁中医药大学附属医院麻醉科,辽宁沈阳110032

出　　处：《世界中西医结合杂志》2023年第10期1982-1988,共7页World Journal of Integrated Traditional and Western Medicine

基　　金：辽宁省教育厅科学技术研究项目(L202002)。

摘　　要：目的观察电针对阿片类药物耐受大鼠机械刺激缩足反应阈值(Paw withdrawal mechanical thresh⁃old,PWMT))、海马组织G蛋白偶联受体(G Protein-Coupled Receptors,GPCRs)及环磷酸腺苷(cyclic AMP,cAMP)、cAMP反应元件结合蛋白(cyclic-AMP response binding protein,CREB)表达的影响,探索G蛋白-谷氨酸受体-cAMP通路是否为电针治疗阿片类药物耐受的可能机制。方法选取60只SPF级SD大鼠,雌雄各半,按随机数字表法分为生理盐水对照组、药物耐受组、谷氨酸受体抑制剂+药物耐受组、电针+药物耐受组及电针+谷氨酸受体抑制剂+药物耐受组,每组各12只。通过对药物耐受组、谷氨酸受体抑制剂+药物耐受组、电针+药物耐受组及电针+谷氨酸受体抑制剂+药物耐受组正常大鼠连续腹腔注射吗啡9 d,2次/d,制备吗啡耐受(Mor⁃phine tolerance,MT)模型,建模成功后,4组继续给予连续7 d腹腔注射吗啡,谷氨酸受体抑制剂+药物耐受组在予腹腔吗啡注射前0.5 h,予腹腔注射谷氨酸受体抑制剂;电针+药物耐受组在予腹腔吗啡注射后0.5 h,予双侧“合谷”“内关”“三阴交”及“足三里”穴位电针干预,断续波,频率2 Hz,2次/d,每次干预时间为10 min,连续7 d;电针+谷氨酸受体抑制剂+药物耐受组给予谷氨酸受体抑制剂和吗啡后,再进行同电针+药物耐受组相同的电针干预。各组分别于造模前、造模第1天、3天、6天、9天和建模成功后第1天、3天、5天、7天检测左足PWMT;采用蛋白质免疫印迹(Western Blot,WB)法检测大鼠海马组织GPCRss、cAMP、CREB蛋白表达,采用酶联免疫吸附测定法(ELISA)检测血浆β-内啡肽(β-EP)含量。结果与生理盐水对照组比较,药物耐受组、谷氨酸受体抑制剂+药物耐受组、电针+药物耐受组及电针+谷氨酸受体抑制剂+药物耐受组大鼠注射吗啡后第1天、3天、6天PWMT明显升高,差异有统计学意义(P<0.01),但注射吗啡9 d后降低至基础水平,与�Objective To investigate the effects of electroacupuncture on paw-withdrawal mechanical threshold(PWMT),and the expression of hippocampal G protein-coupled receptors(GPCRs),cyclic adenosine monophosphate(cAMP)and cAMP response element-binding protein(CREB)in opioid-resistant rats..To explore whether G protein-glutamate receptor-cAMP pathway is involved in the mechanism for electroacupuncture in the treatment of opioid toler⁃ance.Methods Sixty SPF-grade SD rats,half male and half female were selected and divided into 5 groups by using random number table,namely,saline control group(group A),drug-tolerant group(group B),glutamate receptor inhibitor+drug-tolerant group(group C),electroacupuncture+drug-tolerant group(group D)and electroacupuncture+glutamate receptor inhibitor+drug-tolerant group(group E),with 12 rats in each group.The rats in groups B,C,D and E were con⁃tinuously intraperitoneally injected with morphine twice a day for 9 days to prepare a morphine tolerance(MT)model.After successful modeling,the rats in four groups were given morphine intraperitoneally for 7 consecutive days,and the rats in glutamate receptor inhibitor+drug-tolerant group were given intraperitoneal injection of glutamate receptor inhibitor 0.5 h before morphine injection.In the drug-tolerant group,0.5 h after intraperitoneal morphine injection,electroacupuncture intervention was applied on bilateral Hegu Neiguan Sanyinjiao and Zusanli acupoints for 10 min twice a day for 7 consecu⁃tive days,with intermittent wave and frequency of 2 Hz.After glutamate receptor inhibitor and morphine were given in elec⁃troacupuncture+glutamate receptor inhibitor+drug-tolerant group,the same electroacupuncture intervention was per⁃formed on the first day,the 3rd days,the 6th day,and the 9th day before modeling and the first day,the 3 day,the 5th days,and the 7th day after modeling,respectively.Western Blot(WB)was used to detect the GPCRs,cAMP CREB protein expression in the hippocampus of rats,and enzyme-linked immunosorbent assay(ELISA)was use

关 键 词：阿片类药物耐受 电针 机械性痛阈 G蛋白 环-磷酸腺苷 CAMP反应元件结合蛋白 

分 类 号：R285.5[医药卫生—中药学]