Soluble Thrombomodulin Antigen in Conditioned Medium Is Increased by Damage of Endothelial Cells
Soluble Thrombomodulin Antigen in Conditioned Medium Is Increased by Damage of Endothelial Cells
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条件培养基中的可溶性血栓调节蛋白抗原因内皮细胞损伤而增加
DOI：
10.1055/s-0038-1648199
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发表时间：
1991
影响因子：
6.7
通讯作者：
M. Kazama
中科院分区：
文献类型：
--
作者：
H. Ishii;Hiroyuki Uchiyama;M. Kazama
血栓调节蛋白（TM）不仅存在于内皮细胞中，也以可溶性异质片段的形式存在于循环血浆中。研究了内皮细胞释放可溶性TM抗原的机制。培养的人脐静脉内皮细胞在24 h内释放约0.6%的TM抗原进入条件培养液。加入不同浓度（0.01-5.0 μM）的莫能菌素，虽然能抑制纤溶酶原激活物抑制剂-1的分泌，但对TM抗原的释放无影响。当用0.1-1.0 U/ml凝血酶或3 mM二丁酰cAMP刺激后，相对于对照细胞，总细胞TM水平增加1.3或1.4倍时，TM抗原的释放没有增加。内皮细胞暴露于1 μM N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸（FMLP）和100 ng/ml脂多糖（LPS）的混合物中6 h，细胞TM水平相对于对照细胞降低30%，而TM释放不增加。FMLP和LPS刺激的白细胞处理的细胞增加TM抗原释放到介质中的时间依赖性的方式和TM抗原的释放增加的细胞损伤的程度，测定51铬释放。过氧化氢处理的细胞增加TM抗原释放到介质中的时间和浓度依赖性的方式。过氧化氢引起的TM抗原释放的增加也增加了细胞损伤的程度。在还原条件下十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，Western印迹显示未处理的对照细胞条件培养基中的可溶性TM抗原由105、56、48、33、31和28 kDa的6个异质片段组成，而过氧化氢处理的细胞中的可溶性TM抗原由80、56、33、31和28 kDa的5个异质片段组成。31和28 kDa。这些组合物与人血浆中可溶性TM片段的观察结果非常相似。提示可溶性TM抗原不是内皮细胞分泌的，而是细胞损伤的结果。
Summary Thrombomodulin (TM) exists not only in endothelial cells but also in circulating plasma as soluble heterogeneous fragments. A release mechanism of soluble TM antigen from endothelial cells was investigated. Cultured human umbilical vein endothelial cells released about 0.6% of total cellular TM antigen into conditioned medium during 24 h. The release of TM antigen was not influenced by addition of various concentrations (0.01-5.0 μM) of monensin, which inhibits intracellular transport of secretory proteins, though the secretion of plasminogen activator inhibitor-1 from the cells was inhibited. The release of TM antigen was not increased when total cellular TM level increased 1.3- or 1.4-fold relative to control cells after stimulation with 0.1-1.0 U/ml thrombin or 3 mM dibutyryl cAMP, respectively. Exposure of endothelial cells for 6 h to mixture of 1 μM N-formyl-methionyl-leucyl-phenylalanine (FMLP) and 100 ng/ml lipopolysaccharide (LPS) decreased cellular TM level by 30% relative to control cells without increase in the TM release. The FMLP and LPS-stimulated leukocyte treatment of the cells increased the release of TM antigens into the medium in a time-dependent manner and the increased release of TM antigen paralleled the extent of cell damage as measured by 51Cr release. Hydrogen peroxide treatment of the cells increased the release of TM antigens into the medium in a time- and concentration-dependent manner. The increased release of TM antigen by hydrogen peroxide also paralleled the extent of cell damage. Soluble TM antigen in conditioned medium from the untreated control cells was composed of six heterogeneous fragments of 105, 56, 48, 33, 31 and 28 kDa observed on Western blots after sodium dodecyl sulfatepolyacrylamide gel electrophoresis under reducing conditions, and the soluble TM antigen from the hydrogen peroxide-treat-ment cells was composed of five heterogeneous fragments of 80, 56, 33, 31 and 28 kDa. These compositions were quite similar to that observed for soluble TM fragments in human plasma. It is suggested that soluble TM antigen is not secreted from endothelial cells, but results from cellular damage.
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DOI：
10.1073/pnas.83.10.3460
发表时间：
1986-05-01
影响因子：
11.1
作者：
NAWROTH, PP;HANDLEY, DA;STERN, DM
通讯作者：
STERN, DM
影响因子：
15.9
作者：
SMEDLY, LA;TONNESEN, MG;WORTHEN, GS
通讯作者：
WORTHEN, GS
影响因子：
20.3
作者：
K. Moore;C. Esmon;NL Esmon
通讯作者：
K. Moore;C. Esmon;NL Esmon
肿瘤坏死因子增强血管内皮细胞对中性粒细胞介导的杀伤的敏感性。
DOI：
--
发表时间：
1988
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
Varani,J;Bendelow,MJ;Sealey,DE;Kunkel,SL;Gannon,DE;Ryan,US;Ward,PA
通讯作者：
Ward,PA
Fetomodulin：胎儿发育的标记表面蛋白，可由环 AMP 调节。
影响因子：
2.7
作者：
Imada,M;Imada,S;Iwasaki,H;Kume,A;Yamaguchi,H;Moore,EE
通讯作者：
Moore,EE