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作 者:陈天德 曹玉芳[2] CHEN Tiande;CAO Yufang(Department of Critical Care Medicine,Ding'an County People's Hospital,Dingan,Hainan,China,571200;Department of Critical Care Medicine,Haikou Hospital Affiliated to Xiangya School of Medicine,Central South University,Haikou,Hainan,China,570208)
机构地区:[1]定安县人民医院重症医学科,海南定安571200 [2]中南大学湘雅医学院附属海口医院重症医学科,海南海口570208
出 处:《分子诊断与治疗杂志》2022年第8期1329-1333,共5页Journal of Molecular Diagnostics and Therapy
基 金:海南省卫生计生行业科研项目(18A200156)。
摘 要:目的探讨类泛素蛋白人类白细胞抗原F介导转录因子10(FAT10)在心肌细胞缺氧修复损伤中的作用及机制。方法采用H9C2大鼠心肌细胞构建缺氧复氧模型,将心肌细胞分为对照组、缺氧4 h组、复氧2 h组及复氧4 h组。采用细胞活力检测试剂盒(CCK-8)检测细胞活性,采用Annexin V-FITC/PI法检测心肌细胞凋亡,采用微量酶标法检测乳酸脱氢酶(LDH)。将阴性对照RNA序列(si-CON)和FAT10沉默序列转入心肌细胞,将细胞分为对照组、缺氧4 h+si-CON组、缺氧4 h+siRNA组、复氧4 h+si-CON组、复氧4 h+siRNA组。Western-blot检测细胞FAT10、Nrf2、HO-1蛋白表达水平。结果随复氧时间延长,心肌细胞存活率下降,心肌细胞损伤加重,细胞凋亡率升高,在复氧4 h后最低。复氧4 h组细胞存活率低于对照组,而LDH水平及细胞凋亡率高于对照组,差异均有统计学意义(P<0.05)。FAT10蛋白表达水平:复氧4 h+siRNA组<缺氧4 h+siRNA组<缺氧4 h+si-CON组,差异均有统计学意义(P<0.05)。转染siRNA后,心肌细胞FAT10蛋白表达水平下降,凋亡率增高,Nrf2和HO-1蛋白表达水平下降(P<0.05)。结论FAT10可能通过Nrf2/HO-1通路抗心肌细胞凋亡,进而对缺氧修复损伤起到保护作用。Objective To investigate the role and mechanism of ubiquitin-like protein human leukocyte antigen F-mediated transcription factor 10(FAT10)in cardiomyocyte hypoxia-reinjury.Methods H9C2 rat cardiomyocytes were used to construct the hypoxia-reoxygenation model,and the cardiomyocytes were divided into the control group,the hypoxia 4h group,the reoxygenation 2 h group and the reoxygenation 4 h group.Cell viability was detected by cell viability detection kit(CCK-8).Cardiomyocyte apoptosis was detected by Annexin V-FITC/PI method,and lactate dehydrogenase(LDH)was detected by micro-enzyme labeling method.The negative control RNA sequence(si-CON)and FAT10 silencing sequence were transferred into cardiomyocytes,and the cells were divided into the control group,the hypoxia 4 h+si-CON group,the hypoxia 4 h+siRNA group,and the reoxygenation 4h+si-CON group,the Reoxygenation 4 h+siRNA group.Western-blot detection of FAT10,Nrf2,HO-1 protein expression levels.Results With the prolongation of reoxygenation time,the survival rate of cardiomyocytes decreased,the injury of cardiomyocytes aggravated,and the apoptosis rate increased,which was the lowest after 4 h of reoxygenation.The survival rate of cells in the 4 h reoxygenation group was lower than that in the control group,while the level of LDH and the apoptosis rate were higher than those in the control group,and the differences were statistically significant(P<0.05).FAT10 protein expression level:Reoxygenation 4h+siRNA group<hypoxia 4 h+siRNA group<hypoxia 4 h+si-CON group.After transfection of siRNA,the expression level of FAT10 protein in cardiomyocytes decreased,the apoptosis rate increased,and the protein expression levels of Nrf2 and HO-1 decreased(P<0.05).Conclusion FAT10 may resist cardiomyocyte apoptosis through the Nrf2/HO-1 pathway,thereby protecting against hypoxia-reinjury.
关 键 词:人类白细胞抗原F介导转录因子10 心肌 缺氧复氧损伤 核因子E2相关因子2 血红素氧化酶1
分 类 号:R542.2[医药卫生—心血管疾病]
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