SEA0400对大鼠心肌细胞缺血再灌注损伤的保护作用

Protective effects of SEA0400 on ischemia-reperfusion injury in rat myocytes

目的:探讨SEA0400对离体大鼠心肌细胞缺血再灌注损伤的保护作用及其机制。方法:利用酶解法分离成年Wistar大鼠心脏获得心肌细胞悬液,随机分成3组进行研究:(1)对照组:心肌细胞用再灌注液培养60min,未经历缺血再灌注过程;(2)缺血再灌注损伤组:心肌细胞缺血30min,再灌注30min,经历缺血再灌注损伤过程;(3)SEA0400干预组:在心肌细胞缺血再灌注过程中,予以终浓度1μmol/L的SEA0400进行干预。实验结束后测定以上各组心肌细胞内Ca2+浓度和心肌细胞活力。结果:缺血再灌注可导致心肌细胞活力降低和细胞内Ca2+荧光强度显著升高(P<0.01);SEA0400能够明显增加心肌细胞活力和减轻细胞内Ca2+荧光强度升高(P<0.01)。结论:SEA0400可明显减轻缺血再灌注所致心肌细胞内钙超载,维持心肌细胞活力,从而达到心肌保护的作用。

Objective： To study cardioprotective effects and mechanisms of SEA0400 （2- [4- [（2,5- difluorophenyl）methoxy]-5-ethoxy aniline） on ischemia-reperfusion injury in rat myocytes. Methods： Rat myocyte suspension were obtained from adult rat hearts by Langendorff peffusion and enzymatic dissociation,and randomly divided into three groups including the control group, ischemia-reperfusion injury group and SEA0400 intervention group. The control group was incubated in normal condition for 60 minutes. The ischemia-reperfusion injury group underwent ischemia for 30 minutes and reperfusion for 30 minutes respectively. SEA0400 intervention group underwent the same as ischemia-reperfusion injury group in the presence of 1μmol/L SEA（M00. MTF was applied for the measurement of the cardiomyocyte viability at the end of reperfusion period. The intracellular Ca^2＋ concentration was measured by laser scanning confocal microscope （LSCM） and expressed as fluorescence intensity of Fluo-3. Results： Ischemia-reperfusion resulted in the decrease of the cardiomyocyte viability and the increase of the fluorescence intensity of Fluo-3 in rat myocyte. SEA0400 could significantly increase the cardiomyocyte viability and decrease the fluorescence intensity of Fluo-3 in rat myocyte. Conclusion： SEA0400 could attenuate Ca^2＋ overload in cardiomyocytes and improve significantly the cardiomyocyte viability during ischemia and reperfusion periods.