THE EXPRESSION OF MONOCYTE CHEMOTACTIC PROTEIN (MCP-1) IN HUMAN VASCULAR ENDOTHELIUM IN-VITRO AND IN-VIVO
THE EXPRESSION OF MONOCYTE CHEMOTACTIC PROTEIN (MCP-1) IN HUMAN VASCULAR ENDOTHELIUM IN-VITRO AND IN-VIVO
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DOI：
10.1007/bf01772208
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发表时间：
1993-09-08
影响因子：
4.3
通讯作者：
KOLATTUKUDY, PE
中科院分区：
文献类型：
--
作者：
LI, YS;SHYY, YJ;KOLATTUKUDY, PE
单核细胞趋化蛋白（MCP-1）被认为在募集单核细胞到血管内皮中起主要作用，其中单核细胞的粘附是动脉粥样硬化发生的最早事件之一。我们从人主动脉内皮mRNA构建的lambda gt 11 cDNA文库中克隆了MCP-1 cDNA，以检验MCP-1在动脉内皮中的表达是否与其他来源的表达相同。鉴定出与MCP-1相似的670 bp cDNA克隆，序列与其他细胞系相同。以克隆的MCP-1 cDNA为探针进行Northern blot分析，在人主动脉、人肺动脉和人脐静脉内皮细胞培养物中发现了0.68和0.77 kb的两条杂交RNA条带。引物延伸分析表明，两个转录本的大小差异（约为90 bp）并非由于5'-非编码区存在差异。重组IL-1 α（100单位/ml）、IL-1 β（100单位/ml）或tnf - α (200 ng/ml)刺激主动脉内皮细胞中MCP-1转录本的数量显著增加。Northern blot和slot blot分析从新鲜切除的人动脉和静脉的内皮和下壁血管中分离的RNA显示MCP-1转录本。这一观察结果首次表明，MCP-1不仅在人动脉粥样硬化中表达，而且在体内无症状的动脉和静脉中也表达。
A monocyte chemotactic protein (MCP-1) is thought to play a major role in recruiting monocytes to the vascular endothelium where the adherence of monocytes is one of the earliest events in atherogenesis. We cloned MCP-1 cDNA from a lambda gt 11 cDNA library constructed from human aortic endothelial mRNA to test whether MCP-1 expressed in arterial endothelium is identical to those from other sources. A similar to 670 bp MCP-1 cDNA clone was identified and showed the identical sequence with the ones from other cell lines. Northern blot analysis using this cloned MCP-1 cDNA as probe revealed two hybridizing bands of RNA at 0.68 and 0.77 kb in human aortic, human pulmonary arterial, and human umbilical vein endothelial cell cultures. Primer extension analysis showed that the difference in size (similar to 90 bp) between the two transcripts is not due to a difference at the 5'-noncoding region. The amount of MCP-1 transcripts increased dramatically in aortic endothelial cells when stimulated with recombinant IL-1 alpha (100 units/ml), IL-1 beta (100 units/ml), or TNF-alpha (200 ng/ml). Northern blot and slot blot analysis of RNA isolated from both the endothelium and the underlying vessel wall of freshly removed human arteries and veins showed MCP-1 transcripts. This observation demonstrates for the first time that MCP-1 is expressed not only in atherosclerotic human arteries but also in symptom free arteries and veins in vivo.