厌氧培养箱法建立大鼠心肌细胞缺氧/复氧损伤模型  

Establishment of Hypoxia /Reoxygenation Injury Model of Myocardial Cells in Rats by Using Anaerobic Incubator

作　　者：夏中元[1] 王宇[1] 刘菊英[2] 

机构地区：[1]武汉大学人民医院麻醉科,湖北武汉430060 [2]湖北医药学院附属太和医院麻醉科,湖北十堰442000

出　　处：《临床误诊误治》2013年第10期102-104,共3页Clinical Misdiagnosis & Mistherapy

基　　金：国家自然科学基金资助课题(81171783)

摘　　要：目的探讨建立大鼠心肌细胞H9c2缺氧/复氧(hypoxia/reoxygenation,H/R)损伤模型的一种新方法。方法实验使用大鼠心肌细胞H9c2,随机分为对照组和实验组,对照组常规培养不做处理,实验组分为缺氧2、4、8、12 h及缺氧后复氧2 h组。缺氧时换不含血清的低糖DMEM培养基,再置入85%N2、10%H2、5%CO2缺氧环境。缺氧后,加入新鲜DMEM培养基,置入二氧化碳培养箱中继续培养2 h。苔盼蓝染色检测细胞存活率,Annexin V/PI染色行流式细胞术检测细胞凋亡率,分光光度法检测培养基中的乳酸脱氢酶(LDH)含量。结果大鼠心肌细胞经过H/R处理后,与对照组比,细胞存活率显著下降,LDH含量升高,细胞凋亡率显著增加,差异有统计学意义(P<0.05)。结论采用厌氧培养箱法建立大鼠心肌细胞H/R损伤模型简单易行,重复性好。Objective To explore a new way for establishment of a hypoxia/reoxygenation （H/R） injury model of myocar- dial cells line 1-19c2 in rats. Methods Myocardial cells line H9c2 were randomly divided into control group and experimental group. Control group was routinely cultured, while the experimental group was secondly divided into hypoxia for 2, 4, 8 and 12 h groups and posthypoxic reoxygenation for 2 h group. The cells in experimental group were cultured under the condition of 85% N2 , 10% H2 , and 5% CO2 with no-serous low carbohydrates of DMEM in hypoxia. The posthypoxic cells were entered into fresh DMEM and cultured in co2 gas incubator for 2 h. The cell survival rate was detected by Trypan blue exclusion. The apoptotic rate was de- tected by flow cytometry. The content of lactate dehydrogenase （LDH） in the culture medium was detected by spectrophotometric method. Results Compared with those in control group, cell survival rate was significantly reduced, and the LDH content and ap- optotic rate were significantly increased, and the differences were statistically significant （P 〈 0. 05 ）. Conclusion Establishment of H/R injury model in rats by using anaerobic incubator is simple and practicable with good repeatability.

关 键 词：心肌缺血 再灌注损伤 缺氧 复氧 细胞模型 大鼠 

分 类 号：R-332[医药卫生]