MORTM1-TAT对吗啡诱导的大鼠小胶质细胞M1型激活状态的影响  

EFFECT OF MOR^TM1-TAT ON MORPHINE-INDUCED M1-TYPE ACTIVATION OF RAT MICROGLIA

作　　者：郝凤栖 梁永新[1] 范中元 屠后安 张冰 夏婧[1] 褚海辰[1] HAO Fengxi;LIANG Yongxin;FAN Zhongyuan;TU Houan;ZHANG Bing;XIA Jing;CHU Haichen(Department of Anesthesiology, The Affiliated Hospital of Qingdao University, Qingdao 266003, China)

机构地区：[1]青岛大学附属医院麻醉科,山东青岛266003

出　　处：《精准医学杂志》2020年第4期343-346,共4页Journal of Precision Medicine

基　　金：国家自然科学基金项目(8187050711)。

摘　　要：目的研究μ/δ阿片受体异聚体(MDOR)干扰肽(MOR^TM1-TAT)对吗啡诱导的大鼠小胶质细胞M1型激活状态的影响。方法将大鼠小胶质细胞接种于6孔板中,待融合度约60%时,采用随机数字表法分为对照组(A组)、溶剂组(B组)、吗啡组(C组)、吗啡+MOR^TM1-TAT组(D组)4组。A组不进行任何处理,B组加入PBS溶剂,C组加入终浓度为100μmol/L的吗啡溶液,D组加入MOR^TM1-TAT孵育2 h后再加入等剂量吗啡。孵育72 h后,采用细胞免疫荧光技术检测细胞M1表型标志物CD86的表达及MDOR的含量;采用ELISA法检测各组细胞培养上清液中肿瘤坏死因子(TNF-α)、白细胞介素-1(IL-1β)的表达水平。结果与A组相比,B组细胞各项指标表达差异无统计学意义(P>0.05);与B组相比,C组细胞培养上清液中TNF-α、IL-1β及细胞M1表型标志物CD86、MDOR表达水平明显升高(F=104.87~463.50,P<0.05);与C组相比较,D组细胞培养上清液中TNF-α、IL-1β及细胞M1表型标志物CD86、MDOR表达表达水平显著降低(P<0.05)。结论MOR^TM1-TAT通过干扰MDOR形成抑制了吗啡诱导的大鼠小胶质细胞M1型分化及促炎细胞因子的释放,该研究为防治吗啡耐受提供了一定的理论依据。Objective To study the effect ofμ/δopioid receptor heteromeric(MDOR)interfering peptide(MOR^TM1-TAT)on morphine-induced M1-type activation of rat microglia.Methods Rat microglial cells were inoculated into 6-well plates.When the confluence was about 60%,they were divided into control group(group A),vehicle group(group B),morphine group(group C),and morphine+MOR^TM1-TAT group(group D)using a random number table.Group A was given no treatment;group B was given PBS solution;group C was incubated with 100μmol/L morphine solution;group D was incubated with MOR^TM1-TAT for 2 h followed by addition of an equal volume of 100μmol/L morphine solution.After 72 h of incubation,the expression of M1 phenotype marker CD86 and the content of MDOR were measured by immunofluorescence assay;the expression levels of tumor necrosis factor(TNF-α)and interleukin-1β(IL-1β)in cell culture supernatant were determined by ELISA.Results There were no significant differences in the above indices between group A and group B(P>0.05).Compared with group B,group C showed significant increases in the expression of TNF-αand IL-1βin cell culture supernatant as well as the expression of CD86 and MDOR(F=104.87-463.50,P<0.05).Compared with group C,group D showed significant reductions in the expression of TNF-αand IL-1βin cell culture supernatant as well as the expression of CD86 and MDOR(P<0.05).Conclusion MOR^TM1-TAT can atte-nuate morphine-induced M1 phenotype differentiation in rat microglia and proinflammatory cytokine release by interfering with MDOR formation,which provides a theoretical basis for the prevention of morphine tolerance.

关 键 词：小神经胶质细胞 MOR^TM1-TAT 受体 阿片样 δ 受体 阿片样 μ 肿瘤坏死因子α 白细胞介素1β 吗啡 大鼠 WISTAR 体外研究 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R741[医药卫生—基础医学]