[1]陈玉磊,吕风华,陈云玲,等.曲美他嗪对缺氧/复氧心肌细胞焦亡的影响[J].新乡医学院学报,2020,37(1):034-38.[doi:10.7683/xxyxyxb.2020.01.009]
　CHEN Yulei,LYU Fenghua,CHEN Yunling,et al.Effect of trimetazidine on pyroptosis of hypoxia/reoxygenation cardiomyocytes[J].Journal of Xinxiang Medical University,2020,37(1):034-38.[doi:10.7683/xxyxyxb.2020.01.009]
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曲美他嗪对缺氧/复氧心肌细胞焦亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年1
页码:
034-38
栏目:
基础研究
出版日期:
2020-01-05

文章信息/Info

Title:
Effect of trimetazidine on pyroptosis of hypoxia/reoxygenation cardiomyocytes
作者:
陈玉磊; 吕风华; 陈云玲; 尹宏磊; 王　卓; 司澳洋
（新乡医学院第一附属医院心血管内科，河南　卫辉　453100）
Author(s):
CHEN Yulei; LYU Fenghua; CHEN Yunling; YIN Honglei; WANG Zhuo; SI Aoyang
(Department of Cardiology,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)
关键词:
曲美他嗪; 缺氧/复氧; H9C2细胞; 缺血再灌注损伤; 细胞焦亡
Keywords:
trimetazidine; hypoxia/reoxygenation; H9C2 cell; ischemia reperfusion injury; pyroptosis
分类号:
R541R965
DOI:
10.7683/xxyxyxb.2020.01.009
文献标志码:
A
摘要:
目的　探讨曲美他嗪(TMZ)对急性缺氧/复氧（H/R）心肌细胞焦亡的影响。方法　选择大鼠胚胎H9C2心肌细胞为研究对象，将细胞分为正常对照组、H/R组、H/R +50 μmol·L-1 TMZ组、H/R +100 μmol·L-1 TMZ组、H/R+500 μmol·L-1 TMZ组。 H/R组、H/R+50 μmol·L-1TMZ组、H/R+100 μmol·L-1TMZ组、H/R+500 μmol·L-1TMZ组细胞行缺氧12 h/复氧4 h处理模拟大鼠心肌缺血再灌注损伤；H/R+50 μmol·L-1TMZ组、H/R+100 μmol·L-1TMZ组、H/R+500 μmol·L-1TMZ组细胞在构建H/R模型前分别给予50、100、500 μmol·L-1TMZ共孵育1 h；正常对照组细胞常规培养；采用细胞计数试剂盒（CCK8）检测各组心肌细胞活性，乳酸脱氢酶（LDH）试剂盒检测各组心肌细胞培养基中LDH活性，采用实时荧光定量聚合酶链反应法和Western blot法检测心肌细胞中Caspase-1、凋亡相关斑点样蛋白(ASC)和Nod样受体蛋白3（NLRP3）mRNA及蛋白表达水平的变化。结果　与正常对照组比较，H/R组心肌细胞活性降低，LDH活性显著升高,细胞中Caspase-1、NLRP3、ASC mRNA和蛋白表达水平升高（P<0.05）；与H/R组比较，H/R+50 μmol·L-1TMZ 组、H/R+100 μmol·L-1TMZ组、H/R+500 μmol·L-1TMZ组细胞活性显著增加，LDH活性显著下降,细胞中Caspase-1、NLRP3、ASC mRNA和蛋白表达水平降低（P＜0.05）；与H/R+50 μmol·L-1TMZ 组和H/R+500 μmol·L-1TMZ组比较，H/R+100 μmol·L-1TMZ组细胞活性增加，LDH活性下降,细胞中Caspase-1、NLRP3、ASC mRNA和蛋白表达水平降低（P＜0.05）；H/R+50 μmol·L-1TMZ组细胞活性、LDH活性、细胞中Caspase-1、NLRP3、ASC mRNA和蛋白表达水平与H/R+500 μmol·L-1TMZ组比较差异无统计学意义（P>0.05）。结论　TMZ能通过抑制细胞焦亡对H/R的心肌细胞起保护作用；TMZ处理心肌细胞的最适浓度为100 μmol·L-1。
Abstract:
Objective　To investigate the effects of trimetazidine(TMZ)on pyroptosis of acute hypoxia/reoxygenatio（H/R） cardiomyocytes.Methods　The rat embryonic H9C2 cardiomyocytes were selected as subjects,and they were divided into normal control group，H/R group，H/R+50 μmol·L-1 TMZ group，H/R+100 μmol·L-1 TMZ group，H/R+500 μmol·L-1 TMZ group.The cardiomyocytes in the H/R group，H/R+50 μmol·L-1 TMZ group，H/R+100 μmol·L-1 TMZ group and H/R+500 μmol·L-1 TMZ group were given hypoxia for 12 hours and reoxygenation for 4 hours to simulate the myocardial ischemia-reperfusion injury of rats.The cardiomyocytes in the H/R+50 μmol·L-1 TMZ group，H/R+100 μmol·L-1 TMZ group and H/R+500 μmol·L-1 TMZ group were incubated with 50,100,and 500 μmol·L-1 TMZ for one hour before constructing the H/R model.The cardiomyocytes in the normal control group were routinely cultured.The viability of cardiomyocytes was detected by cell counting kit-8.The activity of lactate dehydrogenase (LDH) was detected by lactate dehydrogenase kit.The mRNA and protein expression levels of Caspase-1,apoptosis-associated speck-like protein containing(ASC) and NOD-like receptor pyrin domain containing 3(NLRP3） in the cardiomyocytes were measured by real-time quantitative polymerase chain reaction and western blot.Results　Compared with the control group,in the H/R group the cell viability was lower,the LDH activity was higher and the mRNA and protein expression levels of Caspase-1,ASC and NLRP3 in the cardiomyocytes were higher(P<0.05).Compared with the H/R group,the cell viability was higher,the LDH activity was lower and the mRNA and protein expression levels of Caspase-1,ASC and NLRP3 in the cardiomyocytes were lower in the H/R+50 μmol·L-1 TMZ group，H/R+100 μmol·L-1 TMZ group and H/R+500 μmol·L-1 TMZ group(P<0.05).Compared with the H/R+50 μmol·L-1 TMZ group and the H/R+500 μmol·L-1 TMZ group,the cell viability was higher,the LDH activity was lower and the mRNA and protein expression levels of Caspase-1,ASC and NLRP3 in the cardiomyocytes were lower in the H/R+100 μmol·L-1 TMZ group（P＜0.05）.There was no significant difference in the the cell viability,LDH activity and the mRNA and protein expression levels of Caspase-1,ASC and NLRP3 between the H/R+50 μmol·L-1 TMZ group and the H/R+500 μmol·L-1 TMZ group（P>0.05）.Conclusions　TMZ may protect H/R cardiomyocytes by inhibiting pyroptosis.The optimal concentration of TMZ for cardiomyocytes is 100 μmol·L-1.

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更新日期/Last Update: 2020-01-05