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(Ca MKⅡ)、磷酸化认知缺陷突触相关蛋白(Syn Gap)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Ca MKⅡ、Syn Gap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,Ca MKⅡ、Syn Gap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Ca MKⅡ、Syn Gap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Ca MKⅡ、Syn Gap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。
Objective To investigate the influence of Zuogui Jiangtang Jieyu Fang（ ZJJF） on hippocampus neuron synaptic plasticity related proteins-Ca MKⅡ and Syn Gap 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 Ca MKII and Syn Gap 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 Ca MKII and Syn Gap 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 Ca MKII and Syn Gap were un-regulated in positive drug group and ZJJF group（ P〈 0. 01 or P〈 0. 05）. Conclusion ZJJF had significant regulative effect on Ca MKII and Syn Gap in simulated condition of diabetes complicating depression,which may be one of important mechanisms of ZJJF antagonizing cellular damage and neural degeneration.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2017年第5期392-398,共7页
Journal of Beijing University of Traditional Chinese Medicine
基金
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)~~
作者简介
刘检，男，硕士，助理研究员
通信作者：王宇红，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
