龙牙楤木总皂苷对缺血再灌注损伤大鼠心肌细胞线粒体细胞色素C和膜电位的影响及其作用机制

目的:观察龙牙楤木总皂苷、Mito KATP激活剂二氮嗪(DZ)、Mito KATP抑制剂5-羟基癸酸(5-HD)、龙牙+5-HD对大鼠缺血再灌注损伤(ischemia reperfusion injury,IRI)心肌线粒体细胞色素C和膜电位的影响,并探讨龙牙楤木总皂苷在减轻MIRI中的作用及其机制。方法:将60只雄性Wistar大鼠随机分为对照组(假手术组)、模型组、DZ组、5-HD组、龙牙组和龙牙+5-HD组,每组10只,建立Langendorff离体心脏灌流模型。对照组只穿线不结扎,以改良K-H液持续平衡灌流105分钟;其余5组均穿线结扎冠状动脉停灌30分钟,再分别以改良K-H液、DZ、5-HD、龙牙楤木总皂苷、龙牙+5-HD复灌75分钟(其中龙牙+5-HD组以5-HD复灌15分钟继以龙牙楤木总皂苷复灌60分钟)。提取线粒体,western-blot半定量测定线粒体内细胞色素C水平,荧光酶标仪测线粒体膜电位。结果:龙牙组线粒体细胞色素C释放减少,膜电位较高,与模型组、5-HD组、龙牙+5-HD组相比差异有统计学意义(P0.05),与DZ组相比差异无统计学意义(P0.05)。结论:龙牙楤木总皂苷在MIRI时起到保护作用,能减少MIRI时细胞色素C的释放,稳定线粒体膜电位,其作用可能与mito KATP的开放有关。

Study of Effects and Function Mechanism of Aralosides on Mitochondrial Cytochrome C and Membrane Potential of IRI Rat Cardiac Muscle Cells

Objective:To observe the effects of aralosides,MitoKATP DZX,MitoKATP 5-HD,LongYa plus 5-HD on mitochondrial cytochrome C and membrane potential of IRI rat cardiac muscle cells,and explore the function and mechanism of aralosides on relieving MIRI.Methods:All 60 male wister rats were randomly divided into the control group (sham operation group),the model group,DZ group,5-HD group,aralosides group,aralosides plus 5-HD group,each group 10 rats,Langendorff reperfusion model in isolated heart was built.The control group was given threading,not ligation,persistent and balanced perfusion by improved K-H liquid for 105 rain.The other five groups were given threading and ligation for coronary artery,perfusion stopped for 30 min,then reperfusion by improved K-H liquid,DZ,5-HD,aralosides and aralosides plus 5-HD for 75 rain (the aralosides plus 5-HD group was reperfused for 15 ain,also reperfused by aralosides for 60 min).Mitochondria were extracted,mitochondrial cytochrome C was measured by western-blot semiquantitative analysis.Mitochondrial membrane potential was measured by microplate system.Results:The release of mitochondrial cytochrome C in aralosides group reduced,membrane potential increased,compared with the control group,5-HD group and aralosides plus 5-HD group,there were statistical differences (P＜0.05),compared with DZ group,there were no statistical differences (P＞0.05).Conclusion:Aralosides can protect MIRI,reduce the release of mitochondrial cytochrome C in MIRI,stabilize mitochondrial membrane potential,its function might be related to MitoKATP open.