海马注射miR-181c对CCI模型大鼠热痛的影响  

The effects of intra-hippocampal injection of miR-181c on thermal hyperalgesia of CCI rats

作　　者：杨帆 何丽 徐瑞 徐陶 周涵 刘远贵 刘晓红 曾俊伟 Yang Fan;He Li;Xu Rui;Xu Tao;Zhou Han;Liu Yuangui;Liu Xiaohong;Zeng Junwei(Department of Physiology,Zunyi Medical University,Zunyi 563000,China;School of Clinical Medicine,Zunyi Medical University,Zunyi 563000,China;Key Laboratory of Basic Pharmacology of Ministry of Education,Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学生理学教研室,遵义5630002 [2]遵义医科大学,遵义5630002 [3]遵义医科大学基础药理教育部重点实验室,遵义563000

出　　处：《神经解剖学杂志》2020年第2期161-168,共8页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31860291);2018年贵州省教育厅创新群体重大研究项目(黔教合KY字[2018]025);遵义医科大学大学生创新项目(ZYDC2018072)。

摘　　要：目的:观察大鼠海马CA1区注射miR-18Ic激动剂/抑制剂对坐骨神经缩窄性损伤(CCI)大鼠热痛及海马炎症因子和转录因子表达的影响。方法:成年雄性SD大鼠随机分为6组,分别是假手术组(Sham)、CCI模型组(CCI)、CCI+miR-181c激动剂阴性对照组、CCI+miR-181c抑制剂阴性对照组、CCI+miR-181c激动剂组和CCI+miR-I8le抑制剂组。在术前1 d,术后第1、3、5、7 d测定热缩足潜伏期(TWL)。连续7 d给药后取海马,real time RT-PCR、ELISA及Western Blot检测TLR4、炎症因子(TNF-α和IL-1β)和转录因子(CTCF和Nrt2)表达。结果:与Sham组相比,CCI组大鼠TWL缩短(P<0.05),海马miR-18Ic表达下降(P<0.05),TLR4.TNF-a和IL-1βmRNA表达上调(P<0.05),IL-1β和TNF-α含量增多(P<0.05);与CCI组相比,注射miR-181c激动剂后CCI大鼠TWL延长(P<0.05),海马miR-181c表达上调(P<0.05),而TLR4、TNF-α和IL-IβmRNA表达下降(P<0.05),IL-1β和TNF-c含量下降(P<0.05);注射miR-181c抑制剂则TWL降低(P<0.05),CCI大鼠海马miR-18lc表达进一步下降(P<0.05),TLR4、TNF-α和IL-IβmRNA表达进一步上升(P<0.05),IL-1β和TNF-α含量上升(P<0.05)。与Sham组相比,CCI组海马CTCF表达下降,Nrf2在胞核表达上升(P<0.05)。结论:海马注射miR-181c激动剂减轻CCI大鼠热痛敏可能是由于miR-181e下调TLR4和TNF-α表达导致。CCI大鼠海马miR-181e表达下调可能与CTCF低表达有关。Objective:To investigate the efects of intra-CA1 administration of miR-181c agomir or antagomir on thermnal hyperalgesia and the expression of inflammatory factors and transcription factors in hippocampus of rats with chronie constriction injury(CCI)of the sciaie nerve.Methods:Sprague-Dawley male ralts were randomized into six groups:Sham group,CCI goup,CCI+miR-181c agomir negative control group,CCI+miR-181c antagomir negative control group,CCI+miR-18lc agomir group,CCI+miR-18le antagomir group.Thermal withdrawal latency(TWL)were evaluated before the surgery and on 1,3,5 and 7 day afer surgery.The expression of miR-181c,TLR4,TNF-αand IL-1βmRNA in hippocampus was detected by real time RT-PCR and the content of IL-1β and TNF-αwere detected by ELISA.The expression of CTCF and Nrf2 was detected by Western Blot.Results:TWL in CCI group was signifi-canly lower than that of the sham group(P<0.05).Repeated adnministated of miR-181c agomir atenuate thermal hy-peralgesia(P<0.05)and miR-18le antagomir enhanced thermal hyperalgesia(P<0.05).The level of expression of miR-18lc was lower in hippocampus of CCI rats compare with that of sham rats(P<0.05).The expression of TLR4,TNF-αand IL-I1βmRNA were significantly increased in CCI rats than that of sham group(P<0.05).Compared to CCI group,miR-181c agomir treatment suppressed the expression of TLR4,TNF-αand IL-1βmRNA(P<0.05)but increased the expression of miR-181c(P<0.05)。MiR-181e antagomir treatment suppressed the expression of miR-181c(P<0.05)but increased the expression of TLR4,TNF-αand IL-1βmRNA(P<0.05).Compare to sham group,the expression of CTCF in the eytosol and nucleus of hippocampal issuse were signifcantly decreased in CCI rats,(P<0.05).The expression of Nrf2 in the nucleus of hippocampal tsse was significantly increased in CCI rats(P<0.05).Conclusion:Intra-hippocampal CAI injection of miR-18le agomir ateruale CCl-induced thermal hyperalgesia by inhibiting the expression of TLR4 and TNF-αin hippocampus of rals.CCl-induced the downregulation of miR-18lc may

关 键 词：miR-18lc 神经病理性痛 海马 TLR4 TNF-x 大鼠 

分 类 号：R-332[医药卫生] R651.2