连二亚硫酸钠诱导HT22细胞缺氧复氧模型的制备  

Hypoxia Reoxygenation Model of HT22 Cells Induced by Sodium Hydrosulfite

作　　者：王丹[1,2] 田春艳 陈桂生 亓琪[1] 李海洋[1] 田彩红 WANG Dan;TIAN ChunYan;CHEN GuiSheng;QI Qi;LI Haiyang;TIAN Caihong(Clinical College of Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Craniocerebral Diseases,Yinchuan 750004,China;The Third People’s Hospital of Yinchuan in Ningxia,Yinchuan 750004,China;Department of Neurology,the General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床学院,银川750004 [2]宁夏颅脑疾病重点实验室,银川750004 [3]宁夏银川市第三人民医院,银川7500044 [4]宁夏医科大学总医院神经内科,银川750004

出　　处：《宁夏医科大学学报》2020年第10期983-986,共4页Journal of Ningxia Medical University

基　　金：国家自然科学基金(8156050251)。

摘　　要：目的探讨连二亚硫酸钠(Na2S2O4)诱导小鼠海马神经元细胞(HT22细胞)缺氧/复氧模型的制备方法。方法利用Na2S2O4消除培养基中的氧,模拟细胞缺氧环境。分组培养HT22细胞,给予不同浓度(0.5、1、2、4 mmol·L^-1)的Na2S2O4进行缺氧培养1 h,复氧24 h后,倒置相差显微镜观察细胞形态变化,CCK-8法检测细胞存活率,取细胞培养基上清液检测乳酸脱氢酶(LDH)含量。结果HT22细胞经Na2S2O4缺氧处理1 h,复氧24 h后,与正常对照组相比,随着Na2S2O4浓度的增加,上清液中LDH含量增高(P<0.05),HT22细胞形态损伤加重。结论利用Na2S2O4可以建立细胞缺氧/复氧损伤模型,制备HT22细胞缺氧/复氧模型应选择2 mmol·L^-1 Na2S2O4缺氧1 h、复氧24 h较好。Objective To explore the preparation method of hypoxia/reoxygenation model of HT22 cells(mouse hippocampal neurons)induced by sodium dithionite(Na2S2O4).Methods The anoxic environment of oxygen mimic cells in the medium was eliminated by hyposulphite.HT22 cells were cultured in groups and treated with different concentrations(0.5,1,2,4 mmol·L^-1)of Na2S2O4 for 1 h,after 24 h of reoxygenation,the morphological changes of HT22 cells were observed.The cell survival rate was measured by CCK-8 method,and LDH was measured by cell culture supernatant.Results HT22 cells were treated with Na2S2O4 hypoxia for 1 h and reoxygenation for 24 h,compared with the normal control group,with the increase of sodium hydrosulfite concentration,the higher the LDH content in the supernatant(P<0.05),the morphological damage of HT22 cells was more serious.Conclusion The cell model of hypoxia reperfusion injury can be established by sodium hydrosulfite.It is better to choose 2 mmol·L^-1 Na2S2O4 for 1 h hypoxia and 24 h reoxygenation as the model condition of HT22 cell model.

关 键 词：HT22细胞 连二亚硫酸钠 缺氧模型 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]