过表达解偶联蛋白2抑制氧化应激减轻糖尿病小鼠心肌缺血再灌注损伤
Overexpression of UCP2 alleviates myocardial ischemia-reperfusion injury by inhibiting oxidative stress in diabetic mice
丁佳慧 1吴建江 1程虎 1朱阔 1于文彬 1王江1
作者信息
- 1. 830011 乌鲁木齐, 新疆医科大学第一附属医院麻醉科
- 折叠
摘要
目的:探讨过表达解偶联蛋白 2(UCP2)减轻糖尿病小鼠心肌缺血再灌注损伤(IRI)的机制.方法:60 只 8 周龄C57BL小鼠高脂喂养 6 周后,注射链脲佐菌素建立小鼠糖尿病模型,采用随机数字表法分为 5 组,假手术组(Sham组)、缺血再灌注组(I/R组)、UCP2 抑制剂组(Genipin组)、空载 9 型腺相关病毒(AAV9-NC)组(AN组)和携带UCP2 基因的AAV9 组(AU组),每组 12 只.Sham组仅切开皮肤后缝合,其余 4 组开胸结扎冠状动脉左前降支,缺血 60 min后松开结扎线结,再灌注 120 min建立IRI模型.酶联免疫吸附试验检测血清肌钙蛋白I(cTnI)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平,流式细胞仪观察活性氧(ROS)的产生情况,透射电镜观察线粒体超微结构的变化,Western blot检测UCP2的表达水平,超声心动图评估心功能.结果:与Sham组相比,I/R组、Genipin组、AN组及AU组cTnI、MDA水平明显升高,SOD水平明显下降,ROS产生增多,线粒体结构紊乱,每搏输出量(SV)、射血分数(EF)、左室短轴缩短率(FS)明显降低(P＜0.05).与I/R组相比,Genipin组cTnI、MDA水平明显升高,SOD水平明显下降,ROS产生增多,心肌结构严重受损,线粒体明显肿胀,SV、EF、FS明显降低(P＜0.05).与I/R组相比,AU组cTnI、MDA水平明显降低,SOD水平明显升高,ROS产生减少,心肌结构明显改善,线粒体肿胀减轻,SV、EF、FS的明显升高(P＜0.05);AN组与I/R组cTnI、ROS、MDA、SOD、SV、EF、FS的差异无统计学意义,心肌组织和线粒体损伤程度相似.与Sham组相比,I/R组UCP2 表达水平明显增加(P＜0.05).与I/R组相比,Genipin组UCP2 表达水平明显降低,AU组UCP2 表达水平明显增加(P＜0.05),AN组与I/R组UCP2 表达水平的差异无统计学意义(P＞0.05).结论:过表达的UCP2 通过抑制ROS产生,降低氧化应激水平,改善糖尿病小鼠心肌IRI.
Abstract
Objective:To investigate whether overexpression of uncoupling protein 2(UCP2)alleviates myocardial ischemia-reperfusion injury(IRI)by inhibiting oxidative stress in diabetic mice.Methods:Sixty 8-week-old SPF C57BL mice were fed with high fat diet and injected with STZ to establish a diabetes model.Mice were randomly divided into Sham operation group(Sham group),ischemia-reperfusion group(I/R group),UCP2 inhibitor group(Genipin group),AAV9-NC group(AN9 group)and AAV9-UCP2 group(AU group),with 12 mice in each group.In Sham group,the skin was cut and sutured only.In the other four groups,left anterior descending coronary artery was ligated through thoracotomy,and was then released after 60 minutes of ischemia,followed by 120 minutes of reperfusion to establish the ischemia-reperfusion injury model.Serum levels of cTnI,MDA and SOD were measured by ELISA.The production of reactive oxygen species(ROS)was determined by flow cytometry,and the ultrastructure of mitochondria was observed by transmission electron microscopy.The expression of UCP2 was detected by western blot,and cardiac function was evaluated by echocardiography.Results:Compared with Sham group,cTnI and MDA levels were significantly increased,SOD was significantly decreased,ROS production was increased,mitochondrial structure was disordered,and stroke volume(SV),ejection fraction(EF)and fractional shortening(FS)were significantly decreased in I/R,Genipin,AN and AU groups.Compared with I/R group,cTnI and MDA levels were significantly increased,SOD was significantly decreased,ROS production was increased,myocardial structure was severely damaged with obviously swollen mitochondria,and SV,EF and FS were significantly decreased in the Genipin group(P＜0.05).Compared with I/R group,cTnI and MDA levels were significantly decreased,SOD was significantly increased,ROS production was decreased,myocardial structural changes and mitochondrial swelling were improved,and SV,EF and FS were increased in the AU group(P＜0.05).Compared with Sham group,the expression of UCP2 in I/R group was significantly increased(P＜0.05).Compared with the I/R group,the expression of UCP2 in Genipin group was significantly decreased,and the expression of UCP2 in AU group was significantly increased.Conclusion:Overexpression of UCP2 can reduce oxidative stress reaction by inhibiting ROS production and improve myocardial IRI in diabetic mice.
关键词
解偶联蛋白2/心肌再灌注损伤/糖尿病/氧化应激Key words
Uncoupling protein 2/Myocardial reperfusion injury/Diabetes mellitus/Oxidative stress引用本文复制引用
出版年
2024
NETL
NSTL
维普
万方数据