Inflammatory cytokines stimulated C-reactive protein production by human coronary artery smooth muscle cells
Inflammatory cytokines stimulated C-reactive protein production by human coronary artery smooth muscle cells
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DOI：
10.1161/01.cir.0000096055.62724.c5
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发表时间：
2003-10-21
期刊：
影响因子：
37.8
通讯作者：
Yeh, ETH
中科院分区：
文献类型：
--
作者：
Calabró, P;Willerson, JT;Yeh, ETH
背景 - 在表面健康的男性和女性中，血清C反应蛋白（CRP）水平是心血管事件发生的良好预测指标。一直以来人们认为在急性期反应期间CRP仅由肝细胞产生。有几条证据表明动脉粥样硬化的动脉也可产生CRP。然而，在动脉粥样硬化动脉壁局部产生CRP的细胞类型尚未明确确定。
方法与结果 - 将人冠状动脉平滑肌细胞（HCASMCs）和人脐静脉内皮细胞（HUVECs）与不同浓度的白细胞介素 - 1β（IL - 1β）、白细胞介素 - 6（IL - 6）、它们的组合、肿瘤坏死因子 - α（TNF - α）或脂多糖（LPS）一起孵育。将上清液浓缩，并通过一种对人CRP特异的高灵敏度酶联免疫吸附测定法进行分析。从HCASMCs中提取RNA，使用针对CRP的特异性引物进行逆转录酶 - 聚合酶链反应（RT - PCR）。在与25 ng/mL的IL - 1β和10 ng/mL的IL - 6组合孵育48小时后，在HCASMCs中观察到CRP产生量最大，而单独用IL - 1β或IL - 6孵育仅适度诱导CRP。用TNF - α（50 ng/mL）或LPS（1000 EU/mL）孵育导致CRP产生量的增加与IL - 1β和IL - 6组合相当。通过比较相对CRP mRNA水平的RT - PCR独立证实了HCASMCs中CRP的诱导。然而，在HUVECs中未重现HCASMCs中CRP产生的诱导。
结论 - 这些结果表明，HCASMCs而非HUVECs可在炎症细胞因子的作用下产生CRP。局部产生的CRP可直接参与动脉粥样硬化形成以及心血管并发症的发生。
Background - Serum C-reactive protein (CRP) levels are good predictors of the development of cardiovascular events in apparently healthy men and women. CRP has been believed to be produced exclusively by hepatocytes during the acute-phase response. Several lines of evidence have suggested that atherosclerotic arteries can also produce CRP. However, the cell types that produce CRP locally in the atherosclerotic arterial wall have not been clearly identified.Methods and Results - Human coronary artery smooth muscle cells (HCASMCs) and human umbilical vein endothelial cells (HUVECs) were incubated with interleukin-1beta (IL-1beta), IL-6, their combination, tumor necrosis factor-alpha (TNF-alpha), or lipopolysaccharide (LPS) at different concentrations. The supernatants were concentrated and analyzed by a high-sensitivity enzyme-linked immunosorbent assay specific for human CRP. RNA was extracted from the HCASMCs for reverse transcriptase - polymerase chain reaction (RT-PCR) using specific primers for the CRP. Maximal CRP production was observed in HCASMCs after 48 hours of incubation with the combination of 25 ng/mL of IL-1beta and 10 ng/mL of IL-6, whereas incubation with IL-1beta or IL-6 alone only modestly induced CRP. Incubation with TNF-alpha ( 50 ng/mL) or LPS ( 1000 EU/mL) resulted in an increase in CRP production comparable to the IL-1beta and IL-6 combination. The induction of CRP in HCASMCs was independently confirmed by RT-PCR comparing the relative CRP mRNA levels. The induction of CRP production by HCASMCs was not reproduced in HUVECs, however.Conclusions - These results demonstrated that HCASMCs, but not HUVECs, could produce CRP in response to inflammatory cytokines. The locally produced CRP could directly participate in atherogenesis and the development of cardiovascular complications.