重症失血性休克大鼠血小板线粒体变化及虎杖苷的干预作用  

Changes of platelet mitochondria in rats with severe hemorrhagic shock and intervention effect of polydatin

作　　者：陈云燕[1] 王兴民[1] 宋瑞[1] 赵克森[1] 

机构地区：[1]南方医科大学病理生理学教研室、广东省休克微循环重点实验室,广州510515

出　　处：《中华创伤杂志》2013年第9期882-888,共7页Chinese Journal of Trauma

基　　金：国家自然科学基金资助项目（30672179,30971202）;国家科技重大专项资助项目（2011ZX09101-002-08）;广东省重大科技专项资助项目（2011A08062012）

摘　　要：目的 探讨重症失血性休克大鼠血小板线粒体变化程度与休克轻重程度的关系.方法 选择36只Wistar大鼠并按随机数字表法分为假手术组、休克30,60,120 min组、休克120 min＋等渗盐水＋输血组（NS组）及休克120 min＋虎杖苷＋输血组（PD组）,每组6只.荧光素-荧光素酶试剂盒测定血小板三磷酸腺苷（adenosine triphosphate,ATP）含量；电镜观察血小板线粒体形态；Calcein-AM和CoCl2测定线粒体通透性转变孔状态；JC-1线粒体膜电位试剂盒探测线粒体膜电位（△Ψm）变化；过氧化脂质（lipid hydroperoxide,LPO）试剂盒检测血小板中LPO；吖啶橙（acridine orange,AO）检测血小板溶酶体稳定性. 结果 休克60 min组血小板ATP合成显著减少（P＜0.01）,ATP含量随休克时间增加而减少,在NS组中减少最为明显[为正常值的（50.75±9.15）％].休克30 min组出现线粒体肿胀,嵴结构模糊消失,线粒体膜电位（△Ψm）下降及溶酶体稳定性下降（苍白细胞增多）；休克60 min组钙黄绿素荧光减弱（P＜0.01）.PD组上述各指标都有所改善,特别是血小板ATP水平恢复到正常值的（79.57±8.48）％. 结论 重症休克30～60 min时发生血小板线粒体功能不全,血小板线粒体功能检测可作为重症休克诊断和治疗的一项非损伤性指标.Objective To investigate relationship between the variation degree of platelet mitochondria in rats with severe hemorrhagic shock and the degree of shock.Methods Thirty-six Wistar rats were divided into sham group,shock 30,60,and 120 minutes groups,shock 120 minutes ＋ normal saline （NS） ＋ blood reinfusion group （NS group） and shock 120 minutes ＋ polydatin （PD） ＋ blood reinfusion group （PD group） according to random number table,with six rats per group.Content of ATP in platelets was detected by fluorescein-luciferase assay kit; structure of platelet mitochondria by electron microscope; state of mitochondrial permeability transition pore by Calcein-AM and CoCl2 ; change of mitochondrial membrane potential （△Ψm） by JC-1 mitochondrial membrane potential kit; lipid hydroperoxide （LPO） in platelets by LPO assay kit; stability of platelet lysosomes by acridine orange （AO）.Results ATP released from platelets was reduced significantly in shock 60 minutes group （P 〈0.01） and with the prolong of shock period,further reduction was observed,particularly in NS group [（50.75 ± 9.15）% of normal value].Mitochondrial swelling with poorly defined crista structure,declined △Ψ and low lysosome stability （pale cells were increased） were observed in shock 30 minutes group.Calcein fluorescence in mitochondria was faded in shock 60 minutes group （P 〈 0.01）.Whereas in PD group,all the above indices presented some recovery with ATP level returned to nearly （79.57 ± 8.48） % of normal value in particular.Conclusions Platelet mitochondrial dysfunction takes place at 30-60 minutes following severe shock.Hence,it may be served as an non-invasive index for the diagnose and treatment of severe shock.

关 键 词：休克 出血性 血小板 线粒体 

分 类 号：R285.5[医药卫生—中药学]