摘要
目的 :观察 H2 O2 对心肌细胞内活性氧 (ROS)和游离钙 ([Ca2 + ]i)的影响及褪黑素 (MT)的保护作用。方法 :酶消化法原代分离培养乳鼠心肌细胞 ,分别用荧光探针 DCFH- DA和 Fluo- 3- AM标记 ,H2 O2 和 MT处理后 ,流式细胞仪检测细胞荧光强度的变化。结果 :低浓度的 H2 O2 (10 0 μm ol/ L)随着时间的延长可使心肌细胞内的 ROS呈上升趋势 ,30 m in,6 0 m in与 0 min或对照组相比均明显增高 (P<0 .0 1) ;MT干预后上升趋势均明显减弱 ,在 30 ,6 0 min两组均差异显著 (P<0 .0 1)。 [Ca2 + ]i 亦呈显著上升趋势 ,30 min时与 0 min或对照组相比有差异 (P<0 .0 5 ) ,6 0min时与 30 m in,0 min或对照组相比差异显著 (P<0 .0 1) ;MT干预后其上升明显减少 ,两者差异显著 (P<0 .0 1)。结论 :MT有很好的抗氧化效果 ,能起到保护心肌的作用。
AIM: To investigate the role of H 2O 2 in myocardial intracellular reactive oxygen species (ROS) and free calcium([Ca 2+ ] i)and the effects of melatonin (MT) on them. METHODS: The fluorescent probe DCFH DA was used to detect intracellular levels of ROS and Fluo 3 AM to detect [Ca 2+ ] i. Cultured rat ventricular myocytes were divided into 4 groups: Control group, H 2O 2 group, MT group and MT+H 2O 2 group. Cells were incubated with DCFH DA or Fluo 3 AM prior to the exposure to 100 μmol/L H 2O 2 and/or MT. The changes in cell fluorescence were measured by flow cytometer (FCM) at different time points. RESULTS: Compared with the control group, H 2O 2 at low concentration(100 μmol/L) caused significant time dependent increase in intracellular ROS and [Ca 2+ ] i (ROS:30 min,742±326 vs 200±130, P <0.01, 60 min,1630±421 vs 198±130, P <0.01; [Ca 2+ ] i :30 min,126±114 vs 121±99, P <0.05, 60 min,844±356 vs 121±106, P <0.01). However, those changes were significantly attenuated in MT+H 2O 2 group (ROS:30 min,742±326 vs 223±125, P <0.01, 60 min, 1630±421 vs 317±211, P <0.01; [Ca 2+ ] i : 60 min,844±356 vs 439±261, P <0.01). CONCLUSION: Intracellular ROS and [Ca 2+ ] i increase when cardiomyocytes are challenged by 100 μmol/L H 2O 2 . MT has very good antioxidant effect and can be used to protect cardiomyocytes.
出处
《心脏杂志》
CAS
2001年第1期37-40,共4页
Chinese Heart Journal
关键词
活性氧
心肌细胞
游离钙
褪黑素
实验
reactive oxygen species
cardiomyocyte
intracellular free calcium
melatonin
