TLR4/MyD88/NF-κB信号通路对甲基苯丙胺依赖CPP大鼠海马的影响  

Effect of TLR4/MyD88/NF-κB Signal Pathway on the Hippocampus of Methamphetamine-dependent CPP Rats

作　　者：张园 朱婷娜 曹媛媛 刘鹏亮 王一航 吴亚梅 李利华 赵永娜 洪仕君 ZHANG Yuan;ZHU Tingna;CAO Yuanyuan;LIU Pengliang;WANG Yihang;WU Yamei;LI Lihua;ZHAO Yongna;HONG Shijun(School of Pharmaceutical Sciences&Yunnan Key Laboratory of Pharmacology for Natural Products,Kunming Medical University,Kunming Yunnan 650500,China;School of Pharmaceutical Sciences School of Forensic Medicine,Kunming Medical University,Kunming Yunnan 650500,China;School of Pharmaceutical Sciences School of International Education,Kunming Medical University,Kunming Yunnan 650500,China)

机构地区：[1]昆明医科大学药学院暨云南省天然药物药理重点实验室,云南昆明650500 [2]昆明医科大学法医学院,云南昆明650500 [3]昆明医科大学国际教育学院,云南昆明650500

出　　处：《昆明医科大学学报》2023年第2期7-13,共7页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(81760337);云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目[2019FE00K(-163)]。

摘　　要：目的 研究TLR4/MyD88/NF-κB信号通路对甲基苯丙胺(methamphetamine,MA)依赖的条件性位置偏好(conditioned place preference,CPP)大鼠海马的影响,同时采用特异性抑制剂TAK-242抑制Toll样4受体(Toll-like receptor 4,TLR4),减轻MA依赖诱导的海马神经炎症。方法 建立MA(10 mg/kg,ip,14 d)依赖大鼠CPP模型,分别为生理盐水组、MA组、TAK-242组、MA+TAK-242组。TAK-242组和MA+TAK-242组先分别腹腔注射抑制剂TAK-242(3 mg/kg),1h后MA+TAK-242组再腹腔注射MA(10 mg/kg)。采用Western Blot实验和采用荧光定量PCR实验检测MA依赖CPP大鼠海马中TLR4、MyD88、TRAF6、IκB-α、p-IκB-α、NF-κBp65、p-NF-κBp65蛋白的表达和mRNA表达。结果 与生理盐水组相比,MA组TLR4、MyD88、TRAF6、NF-κBp65的蛋白和mRNA表达均升高(P <0.001或P <0.01),IκB-α的蛋白和mRNA表达下降(P <0.01),p-IκB-α、p-NF-κBp65的表达升高(P <0.01或P <0.05);与MA组相比,MA+TAK-242组TLR4、MyD88、TRAF6、NF-κBp65的蛋白和mRNA表达均下降(P <0.001、P<0.01或P <0.05),IκB-α的蛋白和mRNA表达升高(P <0.01),p-IκB-α、p-NF-κBp65表达下降(P <0.01或P <0.05)。结论 MA依赖可通过激活TLR4/MyD88/NF-κB信号通路,诱导CPP大鼠海马神经炎症的发生,采用特异性TLR4抑制剂可以减轻MA诱导的神经炎症。Objective To study the effect of TLR4/MyD88/NF-κB signal pathway on the hippocampus of methamphetamine(MA) dependent conditioned place preference(CPP) rats,and inhibition of Toll like receptor 4(TLR4) by specific inhibitor TAK-242,thereby reducing MA induced hippocampal neuroinflammation. Methods We established a model of MA(10 mg/kg,ip,14 d) dependent CPP in rats. Rats were randomly divided into 4groups: normal saline group, MA group, TAK-242 group, and MA+TAK-242 group. TAK-242 group and MA+TAK-242 group were intraperitoneally injected with inhibitor TAK-242(3 mg/kg),and MA+TAK-242 group was intraperitoneally injected with MA(10 mg/kg) one hour later. Protein and mRNA expression of TLR4, MyD88,TRAF6,IκB-α,p-IκB-α,NF-κ B p65,p-NF-Κb p65 in hippocampus of MA dependent CPP rats were tested by Western Blot test and fluorescent quantitative PCR,respectively. Results Compared with normal saline group, the expression of protein and mRNA of TLR4, MyD88, TRAF6, NF-κBp65 in MA group increased(P < 0.001 or P < 0.01).The protein of IκB-α and mRNA expression of lactamase decreased(P < 0.01),the expression of p-IκB-α and p-NF-κBp65 increased(P < 0.01 or P < 0.05). Compared with MA group,the protein and mRNA expression of TLR4,MyD88,TRAF6,NF-κBp65 in MA+TAK-242 group decreased(P <0.001,P < 0.01 or P < 0.05). The expression of IκB-α protein and mRNA was increased(P < 0.01),the expression of p-IκB-α and p-NF-κBp65 decreased(P < 0.01 or P < 0.05). Conclusions MA dependency can be achieved by activating TLR4/MyD88/NF-κ B signal pathway,which can induce neuroinflammation on the hippocampus of methamphetamine-dependent CPP rats. The use of specific TLR4 inhibitors can attenuate MA induced neuroinflammation.

关 键 词：甲基苯丙胺依赖 条件性位置偏爱 海马 TLR4/MyD88/NF-κB信号转导通路 

分 类 号：R964[医药卫生—药理学]