ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用

doi:10.12047/j.cjap.5703.2019.013

中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (1): 50-54.doi: 10.12047/j.cjap.5703.2019.013

ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用

柳卓¹,刘检²,凌佳¹,杨琴¹,杨蕙²,孟盼¹,杜青³,赵洪庆¹,王宇红^1,2△

1. 湖南中医药大学, 湖南省中药粉体与创新药物省部共建国家重点实验室培育基地, 长沙 410208;
2. 湖南中医药大学第一附属医院, 长沙 410007;
3. 湖南省中医药研究院, 长沙 412000

收稿日期:2018-05-02 出版日期:2019-01-28 发布日期:2019-06-27
通讯作者: ^△ Tel: 13787028248; E-mail: 573643177@qq.com
基金资助:
国家自然科学基金(81373578)

Effects of ELK-1/JNK/c-Fos on apoptosis of rat hippocampal neurons cultured in vitro with Zuogui Jiangtang Jieyu Formula in simulated diabetes mellitus complicated with depression

LIU Zhuo¹, LIU Jian², LING Jia¹, YANG Qin¹, YANG Hui², MENG Pan¹, DU Qing³, ZHAO Hong-qing¹, WANG Yu-hong^1△

1. Hunan University of Chinese Medicine, Hunan Key Laboratory of Chinese MateriaMedica Powder and Innovative Drugs Established by Provincial and Ministry, Changsha 410208;
2. First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007;
3. Hunan Academy of Chinese Medicine, Changsha 412000, China

Received:2018-05-02 Online:2019-01-28 Published:2019-06-27

摘要/Abstract

摘要： 目的:研究信号通路ELK-1/JNK/c-Fos在左归降糖解郁方(ZGJTJYF)对模拟糖尿病并发抑郁症(DD)环境下抗海马神经元凋亡中的作用。方法:原代培养海马神经元,加入高糖(150 mmol/L)+皮质酮(200 μmol/L),构建DD体外细胞模型;将培养的海马神经元细胞随机分为5组:空白血清组、正常组、左归降糖解郁方含药血清组、阳性药(二甲双胍+氟西汀)含药血清组和模型组(每组3个复孔)。模型组和正常组给予等量培养液,其余组加入相应体积分数10 %的相应血清,均干预18 h。分别采用Hoechst 染色、高内涵细胞成像分析技术和RT-PCR技术分别检测海马神经元凋亡情况及检测凋亡相关ELK-1、JNK和c-Fos 蛋白和基因的表达。结果:与空白组比较,模型组细胞凋亡明显,其海马神经元凋亡数量明显增多,ELK-1、JNK和c-Fos 的mRNA及蛋白表达水平均显著升高(P＜0.05);与模型组比较,左归降糖解郁方含药血清组和阳性药含药血清组大鼠海马神经元可见局部亮点明显减少,凋亡细胞数显著减少,ELK-1、JNK和c-Fos蛋白及mRNA表达均明显下调(P＜0.05),且神经网络及树突连接情况得到明显改善。结论:左归降糖解郁方可以减低DD环境下海马神经元中Elk-1、JNK、c-fos表达而起到抗凋亡的效果。

关键词: 左归降糖解郁方, 糖尿病并发抑郁症, 凋亡通路 , 海马神经元, 大鼠

Abstract: Objective:To study the effects of Zuogui Jiangtang Jieyu Formula (ZGJTJYF, the Chinese Medicine) on hippocampal neuron apoptosis in diabetes mellitus complicated with depression (DD). Methods: The primary cultured hippocampal neurons were treated with high glucose (150 mmol/L) and corticosterone (200 micromol/L) to establish the cell model of DD in vitro. The cultured hippocampal neurons were randomly divided into five groups: blank serum group, normal group, Zuogui Jiangtang Jieyu recipe drug-containing serum group, positive drug (metformin + fluoxetine) drug-containing serum group and model group (three compound holes in each group). The model group and the normal group were given the same amount of culture medium, and the other groups were given the corresponding serum with 10% volume fraction for 18 hours. Hoechst staining, high content cell imaging and RT-PCR were used to detect the apoptosis of hippocampal neurons and the expressions of apoptosis-related ETS-like 1 transcription factor(ELK-1), C-Jun N-terminal kinase(JNK) and c-Fos proteins and genes. Results: Compared with the blank group, the apoptotic number of hippocampal neurons in the model group was increased significantly, and the expression levels of ELK-1, JNK and c-Fos were increased significantly (P＜0.05). Compared with the model group, the local bright spots of hippocampal neurons in the Zuogui Jiangtang Jieyu recipe-containing serum group and the positive drug-containing serum group were decreased significantly, and the number of apoptotic cells was decreased significantly. The expressions of JNK, c-fos protein and mRNA were down-regulated significantly (P＜ 0.05), and the neural network and dendritic junction were improved significantly. Conclusion: Zuo Gui Jiang Tang Jie Yu Formula can reverse the expressions of ELK-1, JNK and c-Fos signals in hippocampal neurons under DD environment and play an anti-apoptotic effect.

Key words: Zuogui Jiangtang Jieyu Formula, diabetes mellitus with depression, apoptosis pathway, hippocampal neurons, rats

柳卓, 刘检, 凌佳, 杨琴, 杨蕙, 孟盼, 杜青, 赵洪庆, 王宇红. ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用[J]. 中国应用生理学杂志, 2019, 35(1): 50-54.

LIU Zhuo, LIU Jian, LING Jia, YANG Qin, YANG Hui, MENG Pan, DU Qing, ZHAO Hong-qing, WANG Yu-hong. Effects of ELK-1/JNK/c-Fos on apoptosis of rat hippocampal neurons cultured in vitro with Zuogui Jiangtang Jieyu Formula in simulated diabetes mellitus complicated with depression[J]. CJAP, 2019, 35(1): 50-54.

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ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用

Effects of ELK-1/JNK/c-Fos on apoptosis of rat hippocampal neurons cultured in vitro with Zuogui Jiangtang Jieyu Formula in simulated diabetes mellitus complicated with depression

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