SIRT1通过CREB/BDNF通路调控吗啡诱导大鼠条件位置偏爱的机制研究  

Effects of SIRT1 on morphine-induced conditioned place preference in rats through CREB/BDNF pathway

作　　者：刘奔 涂婉玉 张腾腾 姚志军[2] 易善勇 赵营[3] 雒国胜 贾文歌 李晨晨 赵斌 魏来 LIU Ben;TU Wan-yu;ZHANG Teng-teng;YAO Zhi-jun;YI Shan-yong;ZHAO Ying;LUO Guo-sheng;JIA Wen-ge;LI Chen-chen;ZHAO Bin;WEI Lai(School of Forensic Medicine,Xinxiang Medical University,Xinxiang Henan 453003,China;School of Basic Medical Science,Xinxiang Medical University,Xinxiang Henan 453003,China;School of Pharmacy,Xinxiang Medical University,Xinxiang Henan 453003,China)

机构地区：[1]新乡医学院法医学院,河南新乡453003 [2]新乡医学院基础医学院,河南新乡453003 [3]新乡医学院药学院,河南新乡453003

出　　处：《中国药理学通报》2023年第7期1263-1270,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.U2004111,81701862);河南省教育厅高校重点科研项目(No.18A310024,18A310025);河南省生物精神病学重点实验室开放课题(No.ZDSYS2018007)。

摘　　要：目的研究腹外侧眶皮层(ventrolateral orbital cortex,VLO)内微量注射sirtuin1(SIRT1)抑制剂EX527对吗啡诱导大鼠条件位置偏爱(conditioned place preference,CPP)的影响,并探讨CREB/BDNF通路在其中的作用。方法应用脑立体定位术在大鼠双侧VLO内留置导管,建立吗啡诱导大鼠CPP模型,d 1为适应,d 2~4为前测,d 5~14为条件性训练,隔日交替腹腔注射吗啡(1 mL·kg^(-1),5 mg·kg^(-1))或生理盐水(1 mL·kg^(-1)),提前30 min通过导管向双侧VLO内微量注射EX527(1μL,5 g·L^(-1))或DMSO(1%,1μL),d 15为测试。CPP测试后立即取脑分离VLO组织,Western blot检测VLO内SIRT1、PSD95、c-fos、p-ERK、CREB、BDNF的表达水平。结果吗啡诱导大鼠CPP模型构建成功,吗啡诱导CPP形成组VLO内SIRT1、PSD95、c-fos、p-ERK、CREB蛋白表达明显增高(P<0.05),BDNF蛋白表达明显降低(P<0.05)。VLO内微量注射EX527抑制了吗啡诱导大鼠CPP形成,且明显降低了VLO内SIRT1、PSD95、c-fos、p-ERK、CREB、BDNF蛋白表达(P<0.05)。结论EX527通过抑制吗啡引起的VLO内SIRT1及其下游相关分子表达的增高,并进一步降低BDNF表达,从而有效阻断了大鼠吗啡成瘾的形成。Aim To investigate the effect of microinjection of EX527,a selective SIRT1 antagonist,into the ventrolateral orbital cortex(VLO)on morphine-induced conditioned place preference(CPP),and to explore the role of CREB/BDNF in it.Methods The cannulas were implanted bilaterally in the VLO of rats by brain stereotaxis surgery, and the model of morphine-induced CPP was established. The behavioral experiment consisted of four stages:habituation (d 1), pre-test (d 2-4), conditioning training (d 5-14) and test (d 15). At the stage of conditioning training, EX527 (1 μL, 5 g·L ^(-1) ) or DMSO (1%, 1 μL) was microinjected into bilateral VLO through the cannulas, and 30 min later, morphine (1 mL·kg ^(-1 ), 5 mg·kg ^(-1) ) or saline was injected intraperitoneally every other day. After the stage of test, the VLO tissue was obtained from brain immediately, and the protein expression level of SIRT1, PSD95, c-fos, p-ERK, CREB and BDNF were detected by Western blot. Results The morphine-induced CPP model was successfully established, and Western blot Results showed that the expression level of SIRT1, PSD95, c-fos, p-ERK and CREB in morphine group were markedly up-regulated ( P <0.05), while BDNF expression level was significantly reduced ( P <0.05). However, after the microinjection of EX527 into the VLO, the formation of morphine-induced CPP in rats was inhibited, and the expression levels of SIRT1, PSD95, c-fos, p-ERK, CREB and BDNF were significantly reduced ( P <0.05). Conclusions Microinjecting EX527 into the VLO could effectively suppress the formation of morphine addiction in rats by inhibiting the increased expression of the SIRT1 and its downstream molecules induced by morphine, and further decreasing the expression of BDNF.

关 键 词：腹外侧眶皮层 SIRT1 EX527 吗啡 成瘾 条件位置偏爱 

分 类 号：R-332[医药卫生] R322.81R338.2R971.2R977.6