高糖诱导大鼠海马神经元细胞模型建立及细胞凋亡的影响  

Establishment of a high glucose-induced rat hippocampal neuronal cell model and the effect of apoptosis

作　　者：许永劼 李程程 朱金凤 朱丽英 高超 代龙光 钱雯 张競之 潘卫 李兴[1,2] XU Yongjie;LI Chengcheng;ZHU Jinfeng(The Affiliated Hospital of Guizhou Medical University,Guizhou Prenatal Diagnosis Center,Guiyang 550004)

机构地区：[1]贵州医科大学附属医院贵州省产前诊断中心,贵州贵阳550004 [2]贵州医科大学医学检验学院,贵州贵阳550004 [3]贵州医科大学环境污染与疾病监控教育部重点实验室,贵州贵阳550004

出　　处：《中风与神经疾病杂志》2019年第1期28-32,共5页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金地区科学基金项目(81560720);国家自然科学基金地区科学基金项目(81560138);贵州省教育厅创新群体重大研究项目(黔教合KY字[2018]021号);贵州省区域内一流学科建设项目-公共卫生与预防医学(黔教科研发2017[85]号)

摘　　要：目的通过不同葡萄糖浓度和不同作用时间诱导SD大鼠海马神经元,建立稳定的糖尿病脑病细胞模型,探讨高糖对海马神经元凋亡情况的影响。方法 (1)海马神经元的原代培养及纯度鉴定。(2)最适高糖浓度探索:CCK-8法检测海马神经元各组细胞活性。(3)最适浓度下高糖作用时间的探索:Western blot检测海马神经元细胞Bcl-2、Bax的表达情况。(4)最适浓度和作用时间下,流式细胞术检测细胞的凋亡率。结果 (1)成功培养海马神经元,神经元细胞经鉴定纯度达85%以上。(2) 45 mmol/L组海马神经元存活率均> 80%。(3) 45 mmol/L-48 h为最适作用时间组,细胞内Bcl-2表达下降、Bax表达升高(P <0. 01)。(4)最适高糖组细胞凋亡率显著升高(P <0. 01)。结论高糖浓度为45 mmol/L、作用时间48 h为理想的糖尿病脑病细胞模型,且高糖通过影响Bcl-2、Bax的表达导致海马神经元凋亡率升高。Objective To induce hippocampal neurons in SD rats by different glucose concentrations and different time of action,and establish a stable cell model of diabetic encephalopathy to investigate the effect of high glucose on hippocampal neuronal apoptosis. Methods( 1) Primary culture and purity identification of hippocampal neurons.( 2) The optimal high glucose concentration was explored;CCK-8 method was used to detect the cell viability of hippocampal neurons.( 3) Exploration of high glucose action time under optimal concentration;Western blot was used to detect the expression of Bcl-2 and Bax.( 4) Apoptosis rate of cells was detected by flow cytometry under optimal concentration and time. Results( 1) The hippocampal neurons were successfully cultured,and the purity of the neurons was 85%.( 2) The survival rate of hippocampal neurons in the 45 mmol/L group was > 80%.( 3) 45 mmol/L-48 h was the optimal time group,the expression of Bcl-2 was decreased and the expression of Bax was increased( P < 0. 01).( 4) The apoptosis rate of the optimal high glucose group was significantly increased( P < 0. 01). Conclusion The high glucose concentration of 45 mmmol/L and the action time of 48 hours are ideal cell models for diabetic encephalopathy,and high glucose can increase the apoptosis rate of hippocampal neurons by affecting the expression of Bcl-2 and Bax.

关 键 词：糖尿病脑病 高糖 海马神经元 模型 细胞凋亡 

分 类 号：R587.2[医药卫生—内分泌]