融合蛋白TAP-SSL5对血小板与粒细胞结合的影响  

Effect of TAP-SSL5 fusion protein on binding of human platelets to neutrophils

作　　者：龚丽莎[1] 房兆飞[1] 胡厚源[1] 刘成海[1] 孟璟[1] 

机构地区：[1]第三军医大学西南医院心血管内科重庆市介入心脏病学研究所,重庆400038

出　　处：《第三军医大学学报》2013年第8期754-758,共5页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划;2009AA02Z115);国家自然科学基金(30871059);国家重大新药创制课题(2013ZX09103003-001)~~

摘　　要：目的探讨融合蛋白TAP-SSL5对血小板与粒细胞结合作用的影响。方法采用CCK-8试剂盒检测TAP-SSL5对细胞活力的影响;流式细胞仪检测THP-1细胞表面CD162(PSGL-1)的表达,及TAP-SSL5对小鼠抗人CD162单抗KPL-1与THP-1细胞结合的抑制作用。以20μmol/L ADP激活人血小板,采用流式细胞仪、瑞氏-姬姆萨染色检测血小板与THP-1细胞或人中性粒细胞的结合情况,并研究TAP-SSL5的干预作用。结果 TAP-SSL5浓度在30 mg/L或以下时对THP-1细胞或中性粒细胞的活力无明显影响。流式细胞仪检测显示,TAP-SSL5(终浓度10 mg/L)能显著抑制KPL-1与THP-1细胞的结合;20μmol/L ADP激活的血小板与THP-1细胞或中性粒细胞的结合率分别为(31.3±8.4)%和(35.6±5.9)%;细胞经10 mg/L TAP-SSL5预先处理后,其结合率分别降至(13.4±6.7)%和(10.4±6.4)%(P<0.01)。瑞氏-姬姆萨染色的结果表明,TAP-SSL5和SSL5均可抑制激活的血小板与中性粒细胞的结合(P<0.05)。结论 TAP-SSL5可通过与PSGL-1的结合,而直接抑制激活的血小板与粒细胞的结合。Objective To investigate the effect of anti-inflammatory and anticoagulant tick anticoagulant peptide（TAP）-staphylococcal superantigen like protein-5（SSL5） fusion protein on the binding of human platelets to neutrophils.Methods Cell viability was assessed by colorimetric cell counting kit-8（CCK-8）.Flow cytometry（FCM） was applied to detect the expression of CD162（PSGL-1） on THP-1 cells as well as the inhibitory effect of TAP-SSL5 on the binding of mouse anti-human CD162 monoclonal antibody（KPL-1） to THP-1 cells.Platelets were activated by 20 μmol/L of ADP,and the binding rates of the activated platelets to THP-1 cells and neutrophils were assayed by FCM and Wright-Giemsa staining,respectively.Results TAP-SSL5≤30 mg/L did not affect the viability of THP-1 cells or human neutrophils,and the TAP-SSL5 at 10 mg/L competitively inhibited KPL-1 binding to THP-1 cells.The binding rates of activated platelets（by 20 μmol/L ADP） to THP-1 cells or neutrophils were（31.3±8.4）% and（35.6±5.9）%,respectively,which decreased to（13.4±6.7）% and（10.4±6.4）% after the THP-1 cells and neutrophils were pre-incubated with 10 mg/L TAP-SSL5（P0.01）.The results of Wright-Giemsa staining showed that both TAP-SSL5 and SSL5 could inhibit the binding of activated platelets to neutrophils（P0.05）.Conclusion TAP-SSL5 can directly inhibit the platelet-neutrophil binding.

关 键 词：金黄色葡萄球菌超抗原样蛋白-5 蜱抗凝血肽 融合蛋白 血小板 中性粒细胞 P-选择素糖蛋白配体-1 

分 类 号：R331.143[医药卫生—人体生理学] R392.12[医药卫生—基础医学]