白藜芦醇保护心肌细胞的信号转导途径研究

目的研究白藜芦醇预处理对心肌缺血再灌注损伤保护作用的信号转导途径。方法培养乳鼠心肌细胞,模拟心肌缺血再灌注损伤,采用四甲基偶氮唑蓝比色法测定细胞活力,检测细胞培养液中乳酸脱氢酶活性。结果心肌缺血再灌注组心肌细胞存活率为（35.7±7.4）%,白藜芦醇处理组心肌细胞存活率为（87.8±4.9）%,与缺血/再灌注损伤组比较有显著性差异（P〈0.01）。NG-硝基-L-精氨酸甲酯、亚甲基蓝、D-鞘氨醇、格列本脲处理心肌细胞30 min后,再以白藜芦醇处理细胞1 h,经历心肌缺血再灌注损伤后心肌细胞存活率分别为（38.3±7.1）%、（46.3±9.7）%、（76.2±6.6）%和（72.2±5.4）%,其中NG-硝基-L-精氨酸甲酯、亚甲基蓝处理组心肌细胞存活率较白藜芦醇组明显减低（P〈0.05）,与心肌缺血再灌注损伤组之间无显著性差异（P〉0.05）,而D-鞘氨醇和格列本脲处理组心肌细胞存活率较白藜芦醇组减低（P〈0.05）,但仍高于心肌缺血再灌注组（P〈0.05）。心肌细胞分别经过NG-硝基-L-精氨酸甲酯、S-亚硝基-乙酞青霉胺处理30 m in后,再以白藜芦醇处理细胞1 h,经历缺血/再灌注损伤,损伤后心肌细胞存活率为（86.4±4.2）%,与缺血/再灌注损伤组比较有显著性差异（P〈0.05）。乳酸脱氢酶活性则与之相反。结论白藜芦醇抑制细胞坏死是通过NO-GC-cGMP依赖途径,而蛋白激酶C的活化和K＋ATP通道的开放可能是其下游重要的信号通路。

The Study on Signal Transduction Pathway of Resvaratrol Pretreatment Effect

Objective To study the signal transduction pathway through which resveratrol pretreatment protects myocardial ischemia-reperfusion induced injury.Methods A cell model of ischemia-reperfusion injury was established.Cell viability was detected by trypan blue transmission method and methyl thiazolyl tetrazolium colorimetry.Lactate dehydrogenase activity in the cell culture fluid was detected.Results The cardiocyte survival rate was（35.7±7.4）% in ischemia-reperfusion group,（87.8±4.9）% in Res pretreated group was significantly different from the ischemia-reperfusion group.After being differently pretreated with N（G）-nitro-L-arginine methyl ester,methylene blue,D-sphingosine,glibenclamide,then pretreated with Res,the cardiocyte survival rate was（38.3±7.1）%,（46.3±9.7）%,（76.2±6.6）% and（72.2±5.4）% while that in N（G）-nitro-L-arginine methyl ester and MB group significantly decreased compared with Res group（P〈0.05）,but without significant difference compared with ischemia-reperfusion injury group.After pretreatment with N（G）-nitro-L-arginine methyl ester and S-nitroso-N-acetylpenicillamine,then with Res,the cardiocyte survival rate was（86.4±4.2）%,similar to ischemia-reperfusion injury group（P〈0.05）.The result of lactate dehydrogenase activity was the opposite.Conclusion The inhibitory effect on necrosis is cGMP-dependent and activation of protein kinase C and opening of（K＋ ATP） channels are important downstream events.