[1]程彦,房兆飞,曲小龙,等.融合蛋白TAP-SSL5对人血小板功能的影响[J].第三军医大学学报,2012,34(06):477-480.
 Cheng Yan,Fang Zhaofei,Qu Xiaolong,et al.Effect of TAP-SSL5 fusion protein on human platelet functions in vitro and in vivo [J].J Third Mil Med Univ,2012,34(06):477-480.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第06期
页码:
477-480
栏目:
论著
出版日期:
2012-03-30

文章信息/Info

Title:
Effect of TAP-SSL5 fusion protein on human platelet functions in vitro and in vivo
作者:
程彦房兆飞曲小龙胡厚源宋治远龚丽莎张静
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
Author(s):
Cheng Yan Fang Zhaofei Qu Xiaolong Hu Houyuan Song Zhiyuan Gong Lisha Zhang Jing
Department of Cardiology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
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糖蛋白GPIbα
Keywords:
staphylococcal superantigen-like protein-5 tick anticoagulant peptide fusion protein platelet glycoprotein Ibα
分类号:
R33-33;R331.143;R341
文献标志码:
A
摘要:
目的      探讨抗炎抗凝双效融合蛋白TAP-SSL5对人血小板功能的影响。      方法      采用健康人富含血小板血浆以凝胶过滤法分离得到人血小板。流式细胞仪检测融合蛋白TAP-SSL5或小鼠抗人CD42b(glycoprotein Ibα, GPIbα)单克隆抗体(HIP1)与血小板的结合情况;以人血小板表面P-选择素的表达及PAC-1的结合反映血小板的激活程度;以全血电阻法定量分析TAP-SSL5对人血小板聚集功能的影响;为评价TAP-SSL5的出血风险,进一步观察了TAP-SSL5对小鼠尾部出血时间的影响。      结果      融合蛋白TAP-SSL5可与血小板结合,并竞争性抑制HIP1与血小板的结合,提示TAP-SSL5可与血小板表面的GPIbα结合,进而抑制GPIbα与vWF的相互作用。但高浓度的TAP-SSL5(终浓度30 mg/L)也能激活血小板,使人血小板表面P-选择素的表达增加(表达阳性率为90.4%)和PAC-1的结合量上升(阳性率为66.3%);终浓度为10、30 mg/L的TAP-SSL5可引起血小板的显著聚集。给小鼠单次尾静脉注射10 mg/kg的TAP-SSL5,可显著延长尾部出血时间,由对照组的(647.1±33.7)s延长至(753.6±127.3)s(P<0.01);而常规剂量组(3 mg/kg TAP-SSL5)尾部出血时间为(612.8±79.1)s,与对照组相比无显著差异(P>0.05)。      结论      融合蛋白TAP-SSL5保留了SSL5与血小板GPIbα结合的功能,有助于进一步提高TAP-SSL5的抗血栓作用,但同时也导致融合蛋白TAP-SSL5在高浓度情况下可延长出血时间和激活血小板。
Abstract:
Objective      To investigate the effect of anti-inflammatory and anticoagulant tick anticoagulant peptide (TAP)-staphylococcal superantigen-like protein 5 (SSL5) fusion protein on human platelet functions.       Methods      Gel-filtered platelets (GFP) were prepared from human platelet-rich plasma (PRP). Flow cytometry (FCM) was applied for the binding assay of TAP-SSL5 fusion protein and mouse anti-human CD42b (GPIbα) monoclonal antibody (HIP1) to platelets. Both the expression of P-selectin on the platelet surface and the PAC-1 binding to the platelets were assayed by FCM. Platelet aggregation induced by TAP-SSL5 fusion protein was analyzed by whole blood impedance platelet aggregometry. The mouse tail bleeding time was recorded after venous injection of TAP-SSL5 fusion protein to evaluate the bleeding risk of TAP-SSL5 fusion protein.       Results      TAP-SSL5 fusion protein could bind to platelets and competitively inhibit HIP1 to bind to platelets, indicating that TAP-SSL5 fusion protein could bind to GPIbα on platelets and inhibit the interaction between GPIbα and von Willebrand Factor (vWF). A high concentration of TAP-SSL5 fusion protein could also activate platelets, and 30 mg/L TAP-SSL5 fusion protein resulted in 90.4% P-selectin positive platelets and 66.3% PAC-1 positive platelets. TAP-SSL5 fusion protein at high concentrations promoted platelets aggregation as well (10 and 30 mg/kg). TAP-SSL5 fusion protein (10 mg/kg) significantly prolonged the mouse tail bleeding time. The mouse tail bleeding time was prolonged to (753.6±127.3)s, which was significantly longer than that of the mice without the injection [(647.1±33.7)s, P<0.01]. However, the tail bleeding time of the conventional dose group (3 mg/kg) was (612.8±79.1)s, which was not significantly different from that of the control group (P>0.05).       Conclusion      TAP-SSL5 fusion protein keeps the ability of SSL5 binding to GPIbα on platelets, which may improve the anti-thrombosis function of TAP-SSL5 fusion protein and also lead to the prolongation of mouse tail bleeding time and platelets activation.

参考文献/References:

程彦, 房兆飞, 曲小龙, 等. 融合蛋白TAP-SSL5对人血小板功能的影响[J].第三军医大学学报,2012,34(6):477-480.

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更新日期/Last Update: 2012-03-22