miR-185对癫痫海马神经元中BDNF-TrkB通路表达的影响

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＋B...

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Published in西安交通大学学报（医学版） Vol. 36; no. 2; pp. 169 - 173
Main Author 蔡浩 李江丽 谢伟 常伟 宋毅军
Format Journal Article
LanguageChinese
Published 天津医科大学总医院 神经内科,天津,300052%天津医科大学总医院 天津医科大学七年制,天津,300052 2015
Subjects
BDNF
miR-185
TrkB
海马神经元
癫痫
蛋白印迹
epilepsy
Western blot
TrkB
miR-185 ?
miR-185?
BDNF
海马神经元
蛋白印迹
癫痫
hippocampal neuron
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ISSN1671-8259
DOI10.7652/jdyxb201502005

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Summary: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miR185。后可以消除对癫痫状态下BDNF／TrkB信号传导通路的抑制。
Bibliography:miR-185 ; BDNF; TrkB; hippocampal neuron; epilepsy; Western blot
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 gr
ISSN:1671-8259
DOI:10.7652/jdyxb201502005