摘要
目的研究缺氧复氧损伤诱导体外培养的新生大鼠的肥大心肌细胞凋亡及干预脂代谢对此过程的作用。方法取体外培养的新生大鼠心肌细胞,以血管紧张素Ⅱ诱导其肥大,分4组,第1组于3%O2、5%CO29、2%N2的三气培养箱中培养12、24、36h,再恢复正常条件培养4h,造成缺氧复氧损伤的肥大心肌细胞模型;第2组加入曲美他嗪(trimetazidine,TMZ)使其终浓度为0.2mM;第3组加入曲美他嗪,使其终浓度为1.0mM;第4组加入曲美他嗪盐,使其终浓度为5.0mM;4组均缺氧复氧相同时间。电镜观察肥大心肌细胞及凋亡细胞的超微结构变化;Hochest33342/PI荧光染色识别凋亡细胞;TUNEL法观察心肌细胞凋亡形态学特征,并计数凋亡心肌细胞数,检测心肌细胞凋亡率。结果肥大心肌细胞在缺氧12、24、36h后复氧4h,TUNEL法可检测到阳性的凋亡细胞,凋亡率分别为:(19.99±4.88)%、(22.66±5.08)%和(24.47±5.74)%;肥大心肌细胞缺氧培养12、24、36h后加入TMZ 0.2mM再复氧4h,检测其凋亡率分别为(19.6±3.02)%(、22.5±3.17)%和(23.7±3.34)%;肥大细胞缺氧培养12、24、36h后加入TMZ 1.0mM再复氧4h,检测其凋亡率分别为(18.6±2.91)%、(20.3±3.02)%和(21.8±2.86))%;肥大心肌细胞缺氧培养12、24、36h后加入TMZ 5.0mM再复氧4h,检测其凋亡率分别为(14.6±2.67)%、(17.3±2.71)%和(20.3±3.28)%。结论缺氧引起的心肌细胞凋亡率随着缺氧时间的延长而呈增高趋势;缺氧复氧较单纯缺氧肥大心肌细胞凋亡率增加;TMZ对缺氧复氧损伤引起的肥大心肌细胞凋亡有抑制作用。
Objective To study the effect of apoptosis of rat hypertrophic cardiomyocytes induced with hypoxia/reoxygenation, and to change metabolic pathway of hypertrophic cardiomyocytes by trimetazidine. Methods （ 1 ） Hypertrophied cardiomyocyte model induced by angiotensin Ⅱ was set up. By [^3H]-Leu incorporation,model of hypertrophied cardiomyocytes was assessed. （2） The cultured media were replaced by low glucose DMEM supplemented with before hypoxia. The cardiomyocytes were incubated at 37C in an air-tighe incubator where normal air was replaced by 92%N2,5%CO2 ,3%O2 for 24h to produce hypoxia,and then the air was replaced by 23 %O2,5% CO2 for 4h to produce reoxygenation. Apoptotic cell death was evaluated using a modified TUNEL （TdT-mediated Dutp Nick-End Labeling）assay（Dead End TM Colorimetric TUNEL）. Results （1）The apoptosis rates of cardiomyoeytes increased with time of hypoxia/reoxygenation（H/R）. The modle for H/R induced apoptosis of culturing rat hypertrophic cardiomyocytes was established successfully. （2）When cardiomyocytes were pretreated with trimetazldine, the rates of apoptosis were decreased. （3） Changing metabolic pathway has effect of apoptosis of rat hypertrophic cardiomyocytes. Conclusions （1）With the method of collagense digestion,it is convenient to obtain primary cultured neonatal Wistar rat ventricular eardiomyocytes. This method is one of vital research tool in the fields of cardiac hypertrophy. （2）By 10-7mol/L concertration,angiotensin Ⅱ could accelerate cellular proliferation, structural protein biosynthesis and cardiac hypertrophy. （3）There is an effect of anti-apoptosis by used trimetazidine.
出处
《重庆医学》
CAS
CSCD
2006年第10期909-911,913,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(30100069)
关键词
肥大心肌细胞
缺氧复氧
凋亡
曲美他嗪
apoptosis ： hypertrophy
hypoxia/reoxygenation cardiomyocyte
trimetazidine
