[1]刘成海,彭松,胡厚源,等.融合蛋白TAP-SSL5对血小板微粒与THP-1细胞结合及Mac-1活化的影响[J].第三军医大学学报,2014,36(09):864-867.
 Liu Chenghai,Peng Song,Hu Houyuan,et al.Fusion protein TAP-SSL5 suppresses binding of platelet microparticles to THP-1 cells and activation of Mac-1[J].J Third Mil Med Univ,2014,36(09):864-867.
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融合蛋白TAP-SSL5对血小板微粒与THP-1细胞结合及Mac-1活化的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第09期
页码:
864-867
栏目:
论著
出版日期:
2014-05-15

文章信息/Info

Title:
Fusion protein TAP-SSL5 suppresses binding of platelet microparticles to THP-1 cells and activation of Mac-1
作者:
刘成海彭松胡厚源贝俊杰孟璟房兆飞
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):
Liu Chenghai Peng Song Hu Houyuan Bei Junjie Meng Jing Fang Zhaofei
Department of Cardiology, Chongqing Institute of Interventional Cardiology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
血小板微粒金黄色葡萄球菌超抗原样蛋白-5蜱抗凝血肽融合蛋白THP-1细胞Mac-1
Keywords:
platelet microparticles staphylococcal superantigen like protein-5 anti-coagulant tick anticoagulant peptide fusion protein THP-1 cells Mac-1
分类号:
R331.143; R341; R392.1
文献标志码:
A
摘要:
目的      探讨融合蛋白TAP-SSL5对血小板微粒(platelet microparticles,PMPs)与人单核细胞株THP-1细胞结合及Mac-1活化的影响。      方法        以二磷酸腺苷(adenosine diphosphate,ADP)激活人血小板并获取PMPs。采用流式细胞仪(flow cytometry,FCM)及PE标记的抗CD62P单克隆抗体、FITC标记的Annexin V检测PMPs,以FITC标记的抗CD41单克隆抗体和PE标记的抗CD154(CD40L)单克隆抗体检测PMPs的表面特征。采用JC-1试剂盒检测血小板线粒体膜电位。采用FCM检测PMPs与THP-1细胞的结合,以及PMPs诱导THP-1细胞表面 Mac-1(CD11b/CD18,αMβ2)的活化情况,并研究TAP-SSL5的干预作用。      结果       PMPs呈现CD62P和Annexin V双阳性,且 CD41和CD40L的阳性率分别达到50.8%和44.0%。JC-1检测显示,ADP对血小板线粒体膜电位无明显影响(P>0.05)。PMPs与THP-1细胞的结合率为(24.80±5.16)%,PMPs 诱导THP-1细胞Mac-1的活化率为(21.17±5.92)%,THP-1细胞经10 mg/L TAP-SSL5预处理后,PMPs的结合率下降至(13.67±2.15)%(P<0.05),Mac-1的活化率下降至(0.99±0.62)%(P<0.01)。      结论        TAP-SSL5可抑制PMPs与THP-1细胞的结合及THP-1细胞表面Mac-1的活化。
Abstract:
Objective       To investigate the effect of anticoagulant and anti-inflammatory fusion protein TAP-SSL5 (tick anticoagulant peptide and staphylococcal superantigen like protein-5) on the binding of platelet microparticles (PMPs) to human acute monocytic leukemia cell line THP-1 and the activation of Mac-1.        Methods        PMPs were generated by adenosine diphosphate (ADP) activating human platelets. Flow cytometry (FCM) was used to identify PMPs by PE labeled mouse anti-human CD62P monoclonal antibody and FITC-Annexin V, and to check the activation and phosphatidylserine (PS) by FITC labeled mouse anti-human CD41 monoclonal antibody and PE labeled mouse anti-human CD151 monoclonal antibody (CD40L). The mitochondrial membrane potential in human platelets was checked with JC-1 kit. The binding rates of PMPs to THP-1 cells and the conformation change of Mac-1 (CD11b/CD18, αMβ2) after co-incubation with PMPs were assayed by FCM.       Results       Both CD62P and PS were positive on PMPs. The positive rates of CD41 and CD40L on the ADP-induced PMPs were 50.8% and 44.0% respectively. While there was no significant change on the mitochondrial membrane potential in ADP activated platelets (P>0.05). The binding rates of PMPs to THP-1 cells and the activation rates of Mac-1 on THP-1 cells were (24.80±5.16)% and (21.17±5.92)% respectively, which decreased to (13.67±2.15)% and (0.99±0.62)% after the THP-1 cells were pre-incubated with 10 mg/L TAP-SSL5 (P<0.05 and P<0.01).       Conclusion       TAP-SSL5 directly inhibits the binding of PMPs to THP-1 cells, and subsequently inhibits the activation of Mac-1 on THP-1 cells.

参考文献/References:

刘成海, 彭松, 胡厚源, 等. 融合蛋白TAP-SSL5对血小板微粒与THP-1细胞结合及Mac-1活化的影响[J].第三军医大学学报,2014,36(9):864-867.

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更新日期/Last Update: 2014-05-05