南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (07): 1075-.

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止血带性肢体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   

  • 出版日期:2014-07-20 发布日期:2014-07-20

Changes of platelet mitochondria in rats with tourniquet-induced limb ischemiareperfusion
injury

  • Online:2014-07-20 Published:2014-07-20

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(P<0.05或P<0.01),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(P<0.05或P<0.01);在12 h时,以上改变有所恢复但仍有统计学差异
(P<0.05或P<0.01);24 h时基本恢复正常(P>0.05)。血小板计数在两组间无统计学差异(P>0.05)。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。

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 hydroperoxide levels in the platelets at 2 and 6 h following the injury (P<0.05 or 0.01). These alterations
recovered partially but remained significantly different from the control levels at 12 h (P<0.05 or 0.01) until full recovery at 24
h. Limb IR injury did not cause significant variations of the platelet counts. Conclusion Tourniquet-induced limb IR injury can
cause mitochondrial damage in the platelets, which occurs mainly in the early stage (6 h) and recovers gradually afterwards
without significant impact on platelet counts.