止血带性肢体缺血再灌注损伤后血小板线粒体的变化.pdf

J South Med Univ, 2014, 34(7): 1075-1078 doi 10.3969/j.issn. 1673-4254.2014.07.34 ·1075 · 基础研究 止血带性肢体止血带性肢体缺血再灌注损伤后血小板线粒体的变化缺血再灌注损伤后血小板线粒体的变化 张建杏，李红英，周国斌，王 庆，李 真 广东省人民医院麻醉科//广东省医学科学院，广东 广州 510080 摘要：目的探讨止血带性缺血再灌注损伤后血小板线粒体的变化，为临床止血带的应用提供指导依据。方法 30 只SD 大鼠随 机分为对照组（6 只），缺血再灌注组（24 只，再随机均分为4 组，分别在再灌注后2、6、12、24 h 取血检测）。采用自制止血带环绕 白兔大腿根部（压力280 mmHg）模拟止血带性缺血，4h 后松开止血带。相应时间点取血分离血小板。采用全自动血球计数仪 测定血小板数量；荧光素—荧光素酶试剂盒测定血小板ATP 含量；JC- 1线粒体膜电位试剂盒探测线粒体膜电位（△ym ）变化；细 胞色素C 凋亡试剂盒检测血小板胞质细胞色素C 含量；过氧化脂质试剂盒检测血小板中过氧化脂质。结果 与对照组比较，缺 血再灌注损伤组血小板内ATP 含量在2、6 h 显著降低（P0.05 或P0.01），而低线粒体膜电位的血小板含量、胞质细胞色素C 含 量、过氧化脂质含量均在2、6 h 逐渐增加并具有统计学差异（P0.05 或P0.01）；在12 h 时，以上改变有所恢复但仍有统计学差异 （P0.05 或P0.01）；24 h 时基本恢复正常（P0.05）。血小板计数在两组间无统计学差异（P0.05）。结论 止血带性缺血再灌注 损伤可以导致血小板线粒体功能的受损，且主要发生在缺血再灌注早期（6 h ），在后期逐渐恢复，而对血小板数量无明显影响。 关键词：止血带；缺血再灌注损伤；线粒体；血小板 Changes of platelet mitochondria in rats with tourniquet-induced limb ischemia- reperfusion injury ZHANG Jianxing, LI Hongying, ZHOU Guobin, WANG Qing, LI Zhen Department of Anesthesiology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China Abstract: Objective To investigate the changes of platelet mitochondria in rats with tourniquet-induced limb ischemia-reperfusion (IR) injury. Methods Thirty SD rats were randomized equally into 5 groups including a control group and 4 limb IR injury groups for blood sampling at 2, 6, 12, or 24 h following IR injury induced by tourniquet on the thighs for 4 h. Platelet was separated from the blood samples for measurement of ATP content, mitochondrial membrane potentials, plasma cytochrome C level, and hydroperoxides. Results Compared with the control group, the rats with tourniquet-induced limb IR injury showed significantly decreased ATP content, lowered mitochondrial membrane potential, and increased plasma cytochrome C and