低温保存对血小板凋亡的影响-内科学专业论文.docx

PAGE PAGE 10 Effection of Low Tempreture on Platelet Apoptosis Abstract Objective To survey the effection of low tempreture on platelet apoptosis. Methods Washed healthy people platelets were stored at 0℃,4℃,10℃,22℃ and 37℃ for up to 5 days. All samples were measured every day.Flow cytometry detected the change of mitochondrial membrane potential. Western blot analysis the apoptosis proteins caspase-3 and antiapoptosis proteins Bcl-xl express. Results The data from flow cytometry indicate that every group exists the change of depolarization of mitochondrial transmembrane potential ( △ Ψm). Compared with other groups, the depolarizations of △Ψm of the group which was stored at 10℃ were inhibited significantly.When stored at 10℃,several apoptoic events in apoptosis including down-regulation of Bcl-xl and caspase-3 activation were inhibited significantly, according to the results of western blot.However, when platelets stored at the other groups, apoptosis protein caspase-3 were activated and expression of anti-apoptosis protein Bcl-xl were decreased. Conclusion These data suggest that low tempreture inhibits platelet apoptosis. The experimental results suggest that attention should be paid to platelet apoptosis under low tempreture conditions such as during platelet storage. Key words:Platelet;apoptosis;low tempreture 引 言 1 细胞凋亡 1.1 细胞凋亡概述 1972 年，Kerr [1]根据细胞的形态学特征首次提出细胞凋亡（apoptosis）。细 胞凋亡又称为细胞程序性死亡（programmed cell death,PCD），是一种主动的、程 序性的由基因控制的细胞自主性死亡方式。细胞凋亡受细胞内源性基因、多种酶 类及信号转导途径的调控，是细胞为更好地适应环境而主动采取的一种死亡过程， 而不是细胞在病理状态下自体损伤的一种现象。细胞凋亡的形态特征是胞质浓缩、 细胞膜内陷并形成凋亡小体、核染色质凝缩及 DNA 大规模的片段化等。越来越 多的证据表明，线粒体是细胞凋亡调控的活动中心，是凋亡的执行者。 Kerr 等[1]发现，衰老及没有功能的有核细胞可出现程序性死亡现象，其能调 节自身细胞的寿命。细胞的凋亡过程可以分为开始阶段及效应阶段。开始阶段包 括线粒体通路介导的内始式途径以及死亡受体通路介导的外始式途径。一旦被激 活 ， 两 条 通 路 可 发 挥 协 同 作 用 ， 激 活 细 胞 内 的 半 胱 天 冬 酶 ( cysteinyl aspar-tate-specific protease,caspase)家族而进入效应阶段，导致细胞程序性死亡， 最终被破坏和吞噬。 1.2 线粒体膜电位改变与细胞凋亡 1890 年，德国生物学家 Altmann 首先发现线粒体这种真核细胞内的细胞器。 线粒体能高效地将储存于有机物中的能量转换成细胞生命活动的直接能源物质三 磷酸腺苷