β_2-GPI与细胞凋亡的关系研究  

Relationship study between the β_2-GPI and apoptosis

作　　者：姜艳芳[1] 谭岩[1] 刘桂英[2] 方艳秋[1] 徐立[3] 刘力华[1] 段秀梅[1] 许淑芬[1] 

机构地区：[1]吉林大学第一医院中心研究室,长春130021 [2]吉林大学第一医院儿科,长春130021 [3]吉林大学第二医院检验科,长春130021

出　　处：《中国免疫学杂志》2006年第2期128-132,共5页Chinese Journal of Immunology

基　　金：国家留学基金委资助(21822067);吉林省科学技术厅科研基金(20030419)资助

摘　　要：目的:探讨β2-糖蛋白-Ⅰ(β2-GPI)与凋亡细胞的结合是否通过磷脂酰丝氨酸(PS)。方法:采用UV照射的方法,诱导K562细胞凋亡,应用碘化丙啶、梯状电泳和β2-GPI检测细胞凋亡。采用β2-GPI与Annexin V的竞争实验来检测β2-GPI是否通过PS与凋亡细胞结合。结果:随着UV照射时间延长,K562细胞的凋亡率增加,在照射300秒时,凋亡细胞达到51.01%(47.23%±4.41%),DNA出现明显的片段化。rGFP连接的β2-GPI,与凋亡细胞结合,β2-GPI可以有效地检测细胞凋亡,而且与Annexin V竞争同一结合位点,即PS。4℃时β2-GPI与晚期凋亡细胞的结合高于37℃,而且随着时间延长,与凋亡细胞的结合均降低。结论:通过β2-GPI与Annexin V的竞争抑制实验第一次为“β2-GPI通过PS与凋亡细胞结合”这一推断提供直接的实验证据,第一次证实β2-GPI既可以与早期凋亡细胞结合,也可以与晚期凋亡细胞结合,rGFP-hβ2-GPI可以作为新的检测细胞凋亡的试剂应用。Objective：To investigate the candidate receptor for β2-GPI on apoptotic cells. Methods ：The apoptotic K562 ceils were induced by Ultra-violet （UV） irradiation and identified by flow cytometry using propidium iodide （PI） staining, Annexin V/PI double staining and DNA fragnatation, respectively. Annexin V/β2-GPI competition experiments were performed to detect FITC- Annexin V binding on the apoptotic cells. Results：Percentage of apoptotic cells reached 51.01% （47.23% -4.41% ） with exposure of the cells to UV 300 s irradiation. It was indicated that recombinant β2-GPI could bind apoptotic cells. The binding resulted in decreased conjuction of labeled annexin V onto the cells in β2-GPI dose-dependent manner. Conclusion： We demonstrate for the first time that 135-GPI is able to bind both early apoptotic cell and late apoptotic cells. β2-GPI may be used as a marker for detection of apoptotlc cells. We speculate that β2-GPI binding on apoptotic cells is mediated by conjugation with phosphatidylserine （PS）, as the pattern of Annexin V＇s.

关 键 词：β2-糖蛋白-Ⅰ ANNEXIN V 细胞凋亡 磷脂酰丝氨酸 

分 类 号：R341[医药卫生—基础医学] R392.11