花生四烯乙醇胺对血小板线粒体细胞色素C的释放和细胞凋亡的影响  

The effects of N-Arachidonoylethanolamine on cytochrome C release and apoptosis of storage platelets

作　　者：庄云龙[1] 李蓬[1] 刘燕[1] 张毅[1] 乔文本 于媛[1] 房云海[1] 徐群[1] 朱传福[1] 张文静[1] 

机构地区：[1]山东省血液中心,山东济南250014

出　　处：《中国输血杂志》2014年第6期589-592,共4页Chinese Journal of Blood Transfusion

基　　金：山东省优秀中青年科学家科研奖励基金(BS2011SW044);山东省医药卫生科技发展计划重点项目(2013WS0170)

摘　　要：目的探讨花生四烯乙醇胺(ANA)对血小板线粒体细胞色素C(Cyt-C)的释放和细胞凋亡的影响。方法将从无偿献血者中采集的血小板,分为实验组(n=20):加入终浓度为0.5μmol/L的ANA;对照组(n=20):未加ANA;2组均置于(22±2)℃水平振荡的振荡仪保存7 d。Western blot检测血小板线粒体Cyt-C的释放和caspase-3的表达、流式细胞仪检测血小板磷脂酰丝氨酸(PS)膜外表达,ELISA方法检测caspase-3的活性。结果实验组与对照组比较,PS表达阳性率:(8.29±1.44)%vs(14.24±2.47)%(P<0.05);caspase-3活性形式表达量:(0.063±0.014)%vs(0.132±0.025)%(P<0.05);caspase-3活性:0.096±0.04 vs 0.208±0.03(P<0.05);Cyt-C从线粒体释放到胞浆的数量比值:(3.18±1.32)%vs(15.13±3.40)%(P<0.05)。结论 ANA降低血板PS的膜外表达、减少线粒体Cyt-C的释放以及降低caspase-3活性,延缓了血小板凋亡。提示ANA可能通过抑制线粒体介导的途径抑制血小板凋亡。Objective To evaluate the effect of NArachidonoylethanolamine（ ANA） on cytochrome C（ Cyt-C）release and apoptosis in platelets（ PLTs）. Methods Apheresis PLTs taken from donors were split into two parts. An aliquot of0. 5 μmol /L ANA final concentration was added to one part of storage PLTs as the experimental group（ n = 20）,the other with-out ANA as the control group（ n = 20）. The two groups samples were stored on a flat-bed shaker at（ 22 ± 2） ℃ up to 7days. The expression of phosphatidyl serine（ PS） positive was determined by flow cytometer. The expressions of Cyt-C and caspase-3 were detected by Western blotting. And caspase-3 activity was measured by ELISA method. Results Comparing the experimental group with the control group,the rates of PLT PS positive were（ 8. 29 ± 1. 44） % vs（ 14. 24 ± 2. 47） %（ P〈 0. 05）; the expressions of activated caspase-3 were（ 0. 063 ± 0. 014） % vs（ 0. 132 ± 0. 025） %（ P〈 0. 05）; the activities of caspase-3 were 0. 096 ± 0. 04 vs 0. 208 ± 0. 03（ P 〈0. 05）; the releases of Cyt-C from mitochondria to cytosol were（ 3. 18± 1. 32） % vs（ 15. 13 ± 3. 40） %（ P〈 0. 05）. Conclusion ANA can inhibit cytochrome C release into the cytoplasm,and reduce caspase-3 activity and PLT apoptosis. This may be one important mechanism of protecting PLT from apoptosis by ANA inhibiting mitochondria-mediated pathway.

关 键 词：花生四烯乙醇胺 血小板 线粒体 细胞凋亡 细胞色素C CASPASE-3 

分 类 号：R446.111[医药卫生—诊断学] R331.143[医药卫生—临床医学]