主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (5): 392-398.doi: 10.3969/j.issn.1006-2157.2017.05.007

• 科技之窗 • 上一篇    下一篇

左归降糖解郁方对糖尿病并发抑郁症海马神经元CaMKⅡ及SynGap的调控作用*

刘检1, 王宇红2#, 韩远山1, 徐雅岚3, 刘林1, 杨蕙1, 李薇1, 向韵2   

  1. 1 湖南中医药大学第一附属医院 湖南 410007;
    2 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;
    3 常德职业技术学院
  • 收稿日期:2016-10-20 出版日期:2017-05-30 发布日期:2017-05-30
  • 通讯作者: 王守红,女,博士,研究员,博士生导师,主要研究方向:中药神经药理学,E-mail:wyh106@126.com.
  • 作者简介:刘检,男,硕士,助理研究员
  • 基金资助:
    *国家自然科学基金项目 (No.81373578, No.81573965),国家自然科学基金青年基金项目(No.81403379, No.81603604),湖南省自然科学基金项目(No.2017JJ3241)

Regulating effect of Zuogui Jiangtang Jieyu Fang on hippocampus neuron synaptic plasticity related proteins-CaMKⅡ and SynGap in simulated condition of diabetes complicating depression*

LIU Jian1, WANG Yuhong2#, HAN Yuanshan1, XU Yalan3, LIU Lin1, YANG Hui1, LI Wei1, XIANG Yun2   

  1. 1 First Affiliated Hospital of Hunan University of Chinese Medicine, Hunan 410007, China;
    2 State Key Laboratory and Breeding Base of Power and Innovative Medicine of Chinese Pharmacy, Hunan 410208, China;
    3 Changde Vocational Technical College, Hunan 415003, China
  • Received:2016-10-20 Online:2017-05-30 Published:2017-05-30
  • Supported by:
    Project sponsored by National Natural Science Foundation (No.81373578, No.81573965), Youth Fund Project sponsored by National Natural Science Foundation (No.81403379, No.81603604), Natural Science Foundation of Hunan Province (No.2017JJ3241)

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)大鼠海马神经元钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)、磷酸化认知缺陷突触相关蛋白(SynGap)的影响。方法 原代分离、纯化和培养SD大鼠海马神经元,采用高糖联合皮质酮构建DD模型。将培养的海马神经元随机分为5组:正常组、模型组、左归降糖解郁方含药血清组、阳性药(氟西汀+二甲双胍)含药血清组和空白血清组。药物干预18 d后,采用尼氏染色检测各组神经元的损伤情况,免疫荧光染色与高内涵细胞成像分析技术(HCA)检测CaMKⅡ、SynGap的表达。结果 与正常组及空白血清组相比,模型组海马神经元内尼氏小体数量显著减少,神经网络、树突棘断裂或消失, CaMKⅡ、SynGap表达明显下调(P<0.01);与模型组相比,阳性药含药血清组与左归降糖解郁方含药血清组尼氏小体明显增加,神经网络、树突数量及突触连接均明显恢复,细胞内CaMKⅡ、SynGap蛋白表达增加(P<0.01或P<0.05)。结论 左归降糖解郁方对DD大鼠模型海马神经元突触可塑性相关蛋白CaMKⅡ、SynGap具有明显的调控作用,这可能是其抗细胞损伤和神经变性的重要机制之一。

关键词: 糖尿病并发抑郁症, 细胞培养, 海马神经元, 左归降糖解郁方, 钙调蛋白依赖性蛋白激酶Ⅱ, 磷酸化认知缺陷突触相关蛋白, 大鼠

Abstract: Objective To investigate the influence of Zuogui Jiangtang Jieyu Fang (ZJJF) on hippocampus neuron synaptic plasticity related proteins-CaMKⅡ and SynGap in rats simulated diabetes complicating depression DD. Methods The hippocampus neurons were primitively isolated, purified and cultured in SD rats. The DD model was established by applying intervention of high glucose and cortisone. The cultured hippocampus neurons were randomly divided into normal group, model group, medicated serum of ZJJF (ZJJF group), medicated serum of positive drug (fluoxetine and metform) group (positive drug group) and blank group. The damages of hippocampus neurons were detected by using Nissl’s staining in all groups, and expressions of CaMKII and SynGap were detected by using immunofluorescence staining and high-content analysis (HCA) after drug intervervention for 18 d. Results Compared with normal group and blank group, the quantity of Nissl bodies decreased significantly, neural network and dentritic spines ruptured or disappeared, and expressions of CaMKII and SynGap were significantly down-regulated in model group (P<0.01). Compared with model group, the quantity of Nissl bodies, neural network and dentritic spines, and synaptic connection were significantly recovered, and expressions of CaMKII and SynGap were un-regulated in positive drug group and ZJJF group (P<0.01 or P<0.05). Conclusion ZJJF had significant regulative effect on CaMKII and SynGap in simulated condition of diabetes complicating depression, which may be one of important mechanisms of ZJJF antagonizing cellular damage and neural degeneration.

Key words: diabetes complicating depression, cell culture, hippocampus neurons, Zuogui Jiangtang Jieyu Fang, calmodulin dependent protein kinase II, SynGap, rats

中图分类号: 

  • R285.5