Xie Chun-Yan, Xiao Jin-Fang, Zhao Zhen-Long
Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China. E-mail:
Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Aug;23(4):1087-91. doi: 10.7534/j.issn.1009-2137.2015.04.035.
To investigate the protective effect of ulinastatin against the activation of tourniquet-induced platelet mitochondria apoptotic signaling.
44 patients with unilateral lower limb operation and tourniquet application were randomly divided into normal saline group and ulinastatin group, and were treated with normal saline and ulinastatin respectively. 12 patents with unilateral lower limb operation but without tourniquet application were enrolled in control group. Lipid hydroperoxide (LPO) in serum was detected by LPO assay kit, the content of ATP was examined by fluorescein-luciferase assay kit; the change of mitochondrial membrane potential (Δ ψm) was detected by JC-1 mitochondrial membrane potential kit; the content of cytoplasmic cytochrome C was examined by Cytochrome C ELISA kit; Caspase-3 activity was detected by Caspase-3 fluorometric assay kit.
As compared with control group, the patients in normal saline group exhibited significant platelet mitochondrial dysfunction which characterized by low ATP level and low mitochondrial membrane potential (Δ ψm) (P < 0.05). Tourniquet application resulted in the activation of the mitochondria apoptotic signaling in platelet, displaying increase in the serum LPO level, release of mitochondrial cytochrome C into the cytoplasm, and activation of caspase-3 (P < 0.05). These alterations above-mentioned were obviously improved by ulinastatin treatment (P < 0.05).
Tourniquet induces platelet mitochondrial dysfunction and mitochondria-dependent apoptotic signaling activation, which can be improved by ulinastatin treatment.
探讨乌司他丁对止血带诱导的血小板线粒体凋亡信号激活的保护作用。
44例接受单侧下肢手术并使用止血带的患者随机分为生理盐水组和乌司他丁组，分别给予生理盐水和乌司他丁治疗。12例接受单侧下肢手术但未使用止血带的患者纳入对照组。采用脂质过氧化（LPO）检测试剂盒检测血清中脂质过氧化物（LPO）水平，采用荧光素-荧光素酶检测试剂盒检测ATP含量；采用JC-1线粒体膜电位试剂盒检测线粒体膜电位（Δψm）的变化；采用细胞色素C ELISA试剂盒检测细胞质细胞色素C含量；采用Caspase-3荧光检测试剂盒检测Caspase-3活性。
与对照组相比，生理盐水组患者血小板线粒体功能明显受损，表现为ATP水平降低和线粒体膜电位（Δψm）降低（P<0.05）。使用止血带导致血小板线粒体凋亡信号激活，表现为血清LPO水平升高、线粒体细胞色素C释放到细胞质中以及Caspase-3激活（P<0.05）。乌司他丁治疗可明显改善上述改变（P<0.05）。
止血带可诱导血小板线粒体功能障碍和线粒体依赖性凋亡信号激活，乌司他丁治疗可改善这种情况。
