米诺环素对OGD/R诱导的H9c2细胞损伤的作用研究  

Effect of minocycline on OGD/R-induced H9c2 cells injury

作　　者：宜全[1] 陈秀会 谭芳慧 刘英华[1] YI Quan;CHEN Xiu-Hui;TAN Fang-Hui;LIU Ying-Hua(Department of Pharmacology of Guangzhou Medical University,Guangzhou 511436,Guangdong Province,China)

机构地区：[1]广州医科大学药理教研室

出　　处：《中国临床药理学杂志》2019年第22期2848-2851,共4页The Chinese Journal of Clinical Pharmacology

基　　金：广东省科技计划基金资助项目(2012B031800235);广州市属高校科研计划基金资助项目(2012C125);广州市科技计划基金资助项目(201707010458);广州市教育局科研课题基金资助项目(1201610286)

摘　　要：目的观察米诺环素对H9c2心肌细胞氧糖剥夺(OGD)/复氧复糖(R)损伤的保护作用,并研究其作用机制是否与抗凋亡有关。方法用减少氧气和糖的供给造成急性心肌细胞缺氧缺糖,然后在预定时间内恢复氧及正常含糖培养基供应的方法建立OGD/R模型。将心肌H9c2细胞分为正常组(常规培养不加干预)、模型组(OGD 6 h/1 h,OGD 6 h/6 h,OGD 6 h/12 h)和低、中、高浓度实验组(OGD/R+1,10和50μmol·L^-1米诺环素)。用台盼蓝和噻唑蓝法检测不同浓度米诺环素和不同时间点复氧对H9c2细胞OGD/R损伤诱导的细胞死亡率或存活率的影响。用Western blotting法检测米诺环素对凋亡相关蛋白Cleaved-Caspase 3、Bax、Bcl-2、JNK及p-JNK的影响。结果在OGD 6 h/R 6 h条件下,模型组细胞死亡率为(30.11±6.16)%,与正常组(1.23%)和中浓度实验组(9.33±2.01)%比较,差异均有统计学意义(均P<0.01)。在OGD 6 h/R 6 h条件下,正常组、模型组和中、高浓度实验组细胞OD值分别为0.35±0.01,0.23±0.02,0.27±0.02和0.28±0.03;在OGD 6 h/R 12 h条件下,正常组、模型组和高浓度实验组细胞OD值分别为0.37±0.01,0.27±0.02和0.32±0.02,2个模型组分别与相应的正常组和中或高浓度实验组比较,差异均有统计学意义(均P<0.05)。在OGD 6 h/R 6 h条件下,正常组、模型组和低、中、高浓度实验组的Cleaved Caspase-3蛋白相对表达量分别为0.83±0.03,1.65±0.24,1.47±0.09,1.29±0.17和0.88±0.16,p-JNK和JNK蛋白相对表达量的比值分别为0.45±0.06,0.84±0.06,0.75±0.05,0.58±0.07和0.37±0.05,模型组的上述指标与正常组和低、中、高浓度实验组比较,差异均有统计学意义(均P<0.05,P<0.01)。在OGD 6 h/R 6 h条件下,正常组、模型组和高浓度实验组的Bax和Bcl-2蛋白相对表达量的比值分别为0.52±0.10,1.00±0.13和0.72±0.18,模型组的上述指标与正常组和高浓度实验组比较,差异均有统计学意义(均P<0.05)。结论米诺环素对H9c2细胞OGD/R损伤�Objective To investigate the protective effect of minocycline on oxygen glucose deprivation(OGD)/reoxygenation(R)-induced H9 c2 cells injury and to study whether its mechanisms is involved in anti-apoptosis.Methods Established OGD/R model:H9 c2 cardiomyocytes were subjected to OGD followed by reoxygenation and the normal feeding medium.H9 c2 cells were divided into normal group(without any intervention),model group(OGD 6 h/1 h,OGD 6 h/6 h,OGD 6 h/12 h)and experimental-L,M,H groups(OGD 6 h/6 h+1,10,50μmol·m L^-1 minocycline,respectively).The cell mortality and viability were detected by Trypan blue and MTT assay at different time points of reoxygenation and in different concentrations.Apoptotic protein cleaved Caspase-3,Bax,Bcl-2,p-JNK and JNK were detected by Western blotting.Results Under the condition of OGD 6 h/R 6 h,model group mortality was(30.11±6.16)%,compared with normal group(1.23±0.36)%and experimental-M group(9.33±2.01)%,the differences were significant(all P<0.05).Under the condition of OGD 6 h/R 6 h,the cell OD values in normal group,model group and experimental-M,H groups were 0.35±0.01,0.23±0.02,0.27±0.02 and0.28±0.03,respectively.Under the condition of OGD 6 h/R 12 h,the cell OD values in normal group,model group and experimental-H groups were 0.37±0.01,0.27±0.02 and 0.32±0.02,respectively.Comparison between normal group and experimental-M or experimental-H group with the corresponding model group,the differences were significant(all P<0.05).Under the condition of OGD 6 h/R 6 h,the cleaved Caspase-3 protein levels in normal group,model group and experimental-L,M,H groups were 0.83±0.03,1.65±0.24,1.47±0.09,1.29±0.17,0.88±0.16,respectively.The ratios of p-JNK/JNK in normal group,model group and experimental-L,M,H groups were 0.45±0.06,0.84±0.06,0.75±0.05,0.58±0.07,0.37±0.05,respectively.Comparison between normal group and the experimental-L,M,H group with the model group,the differences were significant(all P<0.05,P<0.01).The ratios of Bax/Bcl-2 in normal group,model group(OGD

关 键 词：米诺环素 氧糖剥夺 复氧 H9C2细胞 细胞凋亡 

分 类 号：R978.1[医药卫生—药品]