Transcription and translation of human F11R gene are required for an initial step of atherogenesis induced by inflammatory cytokines
Transcription and translation of human F11R gene are required for an initial step of atherogenesis induced by inflammatory cytokines
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DOI：
10.1186/1479-5876-9-98
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发表时间：
2011-06-26
影响因子：
7.4
通讯作者：
Babinska, Anna
中科院分区：
文献类型：
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作者：
Azari, Bani M.;Marmur, Jonathan D.;Babinska, Anna
背景-：F11受体（F11 R; aka JAM-A，JAM-1）是一种细胞粘附蛋白，组成性存在于循环血小板的膜表面和内皮细胞（EC）的紧密连接内。先前的报道表明，内皮细胞暴露于促炎细胞因子导致F11 R分子插入到内皮细胞的腔表面中，随后血小板和内皮细胞的F11 R分子之间发生同源相互作用，并导致血小板粘附到发炎的内皮细胞。本报告的主要新发现是，在这一事件链中的第一步是F11 R分子的从头转录和翻译，通过暴露于炎性细胞因子在EC中诱导。实验方法利用孤立的，将洗涤的人血小板悬浮液和培养的人静脉内皮细胞（HUVEC）和人动脉内皮细胞（HAEC）暴露于促炎细胞因子TNF-α，α和/或IFN-γ，用于检测人血小板通过F11 R粘附于发炎的EC的能力。我们的策略是基于测试以下抑制剂对该活性的影响：一般mRNA合成抑制剂、NF-κ B和JAK/STAT途径的抑制剂以及特异性沉默F11 R基因的小干扰F11 R-mRNA（siRNA）。用抑制剂放线菌素、小白菊素或AG-480处理发炎的EC导致F11 R-mRNA表达的完全阻断，表明NF-κ B和JAK/STAT通路参与了这种诱导。用F11 R siRNA转染EC导致完全抑制了奎宁诱导的F11 R mRNA的上调，并抑制了奎宁炎症EC中新翻译的F11 R分子的检测。F11 R的转录和翻译的抑制的功能后果是显着的人血小板的粘附发炎ECs.Conclusion的阻断：这些结果证明，从头合成的F11 R在内皮细胞是所需的粘附血小板发炎的内皮细胞。由于血小板粘附于发炎的内皮对于非裸露血管中的斑块形成至关重要，因此我们得出结论，F11 R的从头翻译是动脉粥样硬化发生的关键早期步骤，导致动脉粥样硬化，心脏病发作和中风。
Background -: The F11 Receptor (F11R; aka JAM-A, JAM-1) is a cell adhesion protein present constitutively on the membrane surface of circulating platelets and within tight junctions of endothelial cells (ECs). Previous reports demonstrated that exposure of ECs to pro-inflammatory cytokines causes insertion of F11R molecules into the luminal surface of ECs, ensuing with homologous interactions between F11R molecules of platelets and ECs, and a resultant adhesion of platelets to the inflamed ECs. The main new finding of the present report is that the first step in this chain of events is the de-novo transcription and translation of F11R molecules, induced in ECs by exposure to inflammatory cytokines.Methods -: The experimental approach utilized isolated, washed human platelet suspensions and cultured human venous endothelial cells (HUVEC) and human arterial endothelial cells (HAEC) exposed to the proinflammatory cytokines TNF-alpha and/or IFN-gamma, for examination of the ability of human platelets to adhere to the inflamed ECs thru the F11R. Our strategy was based on testing the effects of the following inhibitors on this activity: general mRNA synthesis inhibitors, inhibitors of the NF-kappaB and JAK/STAT pathways, and small interfering F11R-mRNA (siRNAs) to specifically silence the F11R gene.Results -: Treatment of inflamed ECs with the inhibitors actinomycin, parthenolide or with AG-480 resulted in complete blockade of F11R-mRNA expression, indicating the involvement of NF-kappaB and JAK/STAT pathways in this induction. Transfection of ECs with F11R siRNAs caused complete inhibition of the cytokine-induced upregulation of F11R mRNA and inhibition of detection of the newly-translated F11R molecules in cytokine-inflamed ECs. The functional consequence of the inhibition of F11R transcription and translation was the significant blockade of the adhesion of human platelets to inflamed ECs.Conclusion -: These results prove that de novo synthesis of F11R in ECs is required for the adhesion of platelets to inflamed ECs. Because platelet adhesion to an inflamed endothelium is crucial for plaque formation in non-denuded blood vessels, we conclude that the de-novo translation of F11R is a crucial early step in the initiation of atherogenesis, leading to atherosclerosis, heart attacks and stroke.