左归降糖解郁方对模拟糖尿病并发抑郁症环境下模型大鼠海马神经元凋亡相关蛋白的影响

CJI (Traditional Chinese Medicine) - - Chinese Journal Of Information On Traditional Chin -

凌佳1,杨琴 2,刘检 2,王宇红 1,2,徐雅岚 3,韩远山 2,杨蕙 2,孟盼 1

1.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,湖南 长沙 410208;

2.湖南中医药大学第一附属医院,湖南 长沙 410007;

3.常德职业技术学院,湖南 常德 415003

摘要:目的 探讨左归降糖解郁方对模拟糖尿病并发抑郁症(DD)环境下海马神经元凋亡相关蛋白Bcl-2、Bax 及 Caspase-8 的调控作用。方法 采用受孕 18 d SD大鼠胎鼠进行海马神经元原代培养,葡萄糖联合皮质酮干预构建DD模拟环境。将培养的海马神经元随机分为正常组、空白血清组、模型组、阳性药(二甲双胍+氟西汀)药物血清组和待测药(左归降糖解郁方)药物血清组,正常组和模型组给予等量培养液,其余组给予相应体积分数10%药物血清或空白血清,造模干预18 h 后,Hoechst荧光染色检测海马神经元凋亡;高内涵分析技术检测凋亡相关蛋白 Bcl-2、Bax 和 Caspase-8 的表达。结果 与正常组比较,模型组大鼠海马神经元树突

断裂或减少,神经网络连接降低,细胞存在明显的浓染、破碎、光点分布不均现象,Bcl-2蛋白表达显著下降

(P<0.05),Bax 和 Caspase-8 蛋白表达明显升高(P<0.05,P<0.01);与模型组比较,阳性药药物血清组和待测药药物血清组大鼠海马神经元网络连接显著恢复,Hoechst荧光染色可见细胞核明显均匀化,局部亮点明

显减少,Bcl-2 蛋白表达水平显著上调(P<0.05,P<0.01),Bax 和 Caspase-8 蛋白表达水平显著下调(P<0.05,

P<0.01)。结论 左归降糖解郁方对模型大鼠海马神经元凋亡的保护作用可能与调控海马神经元 Bcl-2,Bax和 Caspase-8 的表达有关。

关键词:左归降糖解郁方;糖尿病并发抑郁症;海马神经元;Bcl-2;Bax;Caspase-8;大鼠

DOI:10.3969/j.issn.1005-5304.2017.10.009

中图分类号:R285.5 文献标识码:A 文章编号:1005-5304(2017)10-0035-05

Effects of Zuogui Jiangtang Jieyu Formula on Hippocampus Neuron Apoptosis Related Proteins under Diabetes Mellitus with Depression States

LING Jia1, YANG Qin2, LIU Jian2, WANG Yu-hong1,2, XU Ya-lan3, HAN Yuan-shan2, YANG Hui2, MENG Pan1 (1. Training Bases, Hunan Key Laboratory of Chinese Materia Medica Powder and Innovative Drugs Established by Provincial and Ministry, Changsha 410208, China; 2. First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China; 3. Changde Vocational Technical College, Changde 415003, China)

Abstract: Objective To investigate the regulating effects of Zuogui Jiangtang Jieyu Formula (ZJJF) on Bcl-2, Bax and Caspase-8 in hippocampus neuron damage in diabetes mellitus with depression (DD). Methods Hippocampal neurons from 18 d pregnant rats were primitively cultivated, and then combination of glucose and corticosterone was used to construct DD simulation environment. Cultivated hippocampal neurons were randomly divided into normal group, blank serum group, model group, positive medicine (metformin+fluoxetine) serum group and to-be-tested medicine (ZJJF) serum group. Normal group and model group were given same amount culture medium, while other group were given relevant amont of 10% medicine serum or blank serum. After modeling intervention for 18 h,

基金项目:国家自然科学基金(81373578、81573965、81403379、81403387)

通讯作者:王宇红,E-mail:573643177@qq.com

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