miR-185对癫痫海马神经元中BDNF-TrkB通路表达的影响  

Effect of miR-185* on BDNF/TrkB signaling pathway expression in epileptic neurons

作　　者：蔡浩[1] 李江丽[2,3] 谢伟[1] 常伟[2,3] 宋毅军[1] 

机构地区：[1]天津医科大学总医院神经内科,天津300052 [2]天津医科大学总医院 [3]天津医科大学,七年制天津300052

出　　处：《西安交通大学学报（医学版）》2015年第2期169-173,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金(No.91132722,81071044)~~

摘　　要：目的研究海马神经元癫痫模型中的BDNF/TrkB信号通路及miR-185*对其的调控。方法构建miR-185*表达载体。体外培养海马神经元,7d后将其随机分为正常组、癫痫组、正常+BDNF组、癫痫+BDNF组、正常转染miR-185*组、癫痫转染miR-185*组、癫痫转染miR-185*+BDNF组。免疫荧光、膜片钳及免疫印迹技术观察转染miR-185*后BDNF/TrkB通路的表达变化。结果癫痫+BDNF组海马神经元细胞的TrkB蛋白磷酸化水平较正常+BDNF组降低,有统计学意义(P<0.05);正常+BDNF组TrkB蛋白的磷酸化水平比正常组的磷酸化水平升高,有统计学意义(P<0.001);癫痫+BDNF组TrkB蛋白磷酸化水平比癫痫组升高,有统计学意义(P<0.05);而癫痫+miR-185*+BDNF组的TrkB蛋白磷酸化水平较癫痫+BDNF组和癫痫转染miR-185*组升高(P<0.001)。结论BDNF可以激活BDNF/TrkB信号通路,转染miR-185*后可以消除对癫痫状态下BDNF/TrkB信号传导通路的抑制。Objective To study the BDNF/TrkB signaling pathway in the epilepsy model of hippocampal neurons and the regulatory effect of on it. Methods The primary hippocampal neurons cultured in vitro for 7 days were randomly divided into seven groups： control group, epilepsy group, control＋BDNF group, epilepsy＋BDNF group, control--miR-185 group, epilepsy＋miR-185 group, and epilepsy＋ miR-185-F BDNF group. We constructed miR-185＊ lentivirus vector and observed the changes of BDNF/TrkB pathway expression after transfaction of miR-185 by immunohistochemistry, patch clamp technique and Western blot technique. Results Compared with the control＋BDNF group,the phosphorylated TrkB （pTrkB）/TrkB value was significantly lower in epilepsy＋BDNF group （P〈 0. 05） and control group （P 〈 0. 001）. Compared with the epilepsy group, the phosphorylated TrkB （pTrkB）/TrkB value was significantly higher in epilepsy--BDNF group （P〈0.05）. Compared with the epilepsy＋miR-185 -FBDNF group, the phosphorylated TrkB （pTrkB）/TrkB value was significantly lower in epilepsy-F BDNF group and epilepsy＋miR-185 group （P 〈 0. 001）. BDNF could promote the signaling conduction and miR-185. could remove the inhibition of BDNF/TrkB signaling. Conclusion BDNF can activate the BDNF/TrkB signaling pathway and transfection with miR-185 can relieve the inhibition of BDNF/TrkB signaling pathway of epileptic state by up-regulating the expression of TrkB.

关 键 词：miR-185 BDNF TRKB 海马神经元 癫痫 蛋白印迹 

分 类 号：R741.02[医药卫生—神经病学与精神病学]