两种方法提取乳鼠神经细胞建立缺氧/复氧损伤模型的研究  

Establishment of hypoxia/reoxygenation injury model by extraction of neural cells from neonatal rat by two methods

作　　者：刘威[1] 王爽 温娜 王洪羽[1] 金宏[1] LIU Wei;FANG Shuang;WEN Na(Jinlin Medical College, Jilin 132013, China)

机构地区：[1]吉林医药学院临床医学院临床技能实训中心,吉林吉林132013 [2]吉林医药学院基础医学院形态学教研室,吉林吉林132013

出　　处：《中风与神经疾病杂志》2018年第4期292-295,共4页Journal of Apoplexy and Nervous Diseases

基　　金：吉林省教育厅"十二五"科学技术研究规划项目[吉科教合字(2015)第408号]

摘　　要：目的比较两种不同方法培养乳鼠神经细胞稳定性,并探讨H_2O_2对原代培养神经细胞造成缺氧/复氧损伤模型的最佳处理时间。方法两种方法培养原代神经细胞,加入600μmol/L H_2O_2培养50 min、2 h、3 h后换正常培养液继续培养18 h,造成神经细胞缺氧/复氧损伤。MTT法检测各组细胞生存率,检测LDH活性、SOD、MDA含量和RhoA活性。结果与空白组相比,缺氧50 min、2 h、3 h培养各组细胞存活率下降,LDH活性增加,SOD含量下降,MDA含量升高,RhoA含量增加P均<0.05。与直接法组相比,种植法组细胞在缺氧50 min、2 h各组细胞存活率高,细胞LDH漏出少,SOD含量高,MDA生成少P均<0.05。结论与直接法相比,种植法培养的神经细胞状态更稳定,缺氧50 min、复氧18 h时制作神经细胞损伤模型最接近体内缺氧损伤状态,可用于实验。Objective To compare the stability of primary cultured rat neural cells by two different methods,and to investigate the optimal treatment time of hypoxia/reoxygenation injury model induced by H2O2 in primary cultured neural cells. Methods Primay rat neural cells well cultured Cultivation by two methods： direct method and planting method of two methods of primary rat neural cells,were 600 mol/L H2O2 were added into culture medium for 600 mol/L H2O2 50 min,2 h training and,3 h separately to cause nerve cells injury in hypoxia/reoxygenation,after then changing changed back to the normal culture medium to culture 18 h,caused by nerve cells in hypoxia/reoxygenation injury. MTT assay was used to detect cell viability,LDH activity,intracellular SOD,MDA content and RhoA activity in the supernatant were also detected. Results Compared with the blank group,the cell viability decreased,the LDH activity increased,the intracellular SOD content decreased,the MDA content increased,and the RhoA content increased,P〈0. 05 in the hypoxia 50 min,2 h and 3 h groups.Compared with the direct method group,the cell survival rate of the cells in the hypoxia groups 50 min,2 h was were higher,the cell LDH leakage was less,the SOD content was higher,and the MDA formation was less,P〈0. 05. Conclusion Compared with the direct method,the nerve cells planting culture method is more stable,600 mol/L H2O2 50 min 18 h hypoxia reoxygenation,making nerve cell injury model is closest to the damage state in vivo hypoxia,it can be used for the experiment.

关 键 词：原代培养神经细胞 缺氧/复氧损伤 超氧化物歧化酶 丙二醛 RHOA 

分 类 号：R743.3[医药卫生—神经病学与精神病学]