TNF-alpha decreases expression of somatostatin, somatostatin receptors, and cortistatin in human coronary endothelial cells.
TNF-alpha decreases expression of somatostatin, somatostatin receptors, and cortistatin in human coronary endothelial cells.
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DOI：
10.1016/j.jss.2004.07.244
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发表时间：
2005-02
期刊：
影响因子：
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通讯作者：
Shaoyu Yan;Min Li;H. Chai;Hui Yang;P. Lin;Q. Yao;Changyi J. Chen
中科院分区：
文献类型：
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作者：
Shaoyu Yan;Min Li;H. Chai;Hui Yang;P. Lin;Q. Yao;Changyi J. Chen
材料与方法人冠状动脉内皮细胞（human coronary artery endothelial cells，HCAECs）培养24 h后，加入TNF-α（0.1，1，10 ng/ml），观察培养液中生长抑素（somatostatin，SST）及其受体（somatostatin，SSTRs）的表达及TNF-α对其表达的调节作用。通过实时RT-PCR测定SST、SSTR-1-5以及管家基因（β-actin）的mRNA水平。SSTR-2的表达也通过免疫荧光染色证实。通过[3 H]胸苷掺入法检测SST处理（0.04、0.2或1 ng/ml）对细胞增殖的影响。SSTR-2的mRNA表达水平高于SSTR-1和SSTR-5。然而，SSTR-3和SSTR-4不表达或极低表达。经TNF-α处理后，SST、SSTR-1、SSTR-2和SSTR-5的mRNA水平显著降低，且呈剂量依赖性。与对照组相比，TNF-α（1 ng/ml）使SST、SSTR-1、SSTR-2和SSTR-5分别降低93%、51%、85%和99%（P < 0.001，t检验）。TNF-α处理后SSTR-2的免疫反应性也降低。SST处理的细胞表现出显着减少[3 H]胸苷掺入的剂量依赖性的方式。结论：TNF-α处理后，SHCAEC表达SST、SSTR-1、SSTR-2和SSTR-5均降低。此外，外源性SST处理显着降低细胞增殖，并且这种抑制作用也被TNF-α减弱。
BACKGROUNDThe objective of this study was to determine the expression of somatostatin (SST) and its receptors (SSTRs) and their regulation by TNF-α as well as cell proliferation in response to SST in human endothelial cells.MATERIALS AND METHODSHuman coronary artery endothelial cells (HCAECs) were cultured without or with TNF-α (0.1, 1, or 10 ng/ml) for 24 h. The mRNA levels of SST, SSTR-1-5, as well as a housekeeping gene (β-actin) were determined by real-time RT-PCR. Expression of SSTR-2 was also demonstrated by immunofluorescence staining. Cell proliferation in response to SST treatment (0.04, 0.2, or 1 ng/ml) was performed by [3H]thymidine incorporation.RESULTSWithout TNF-α treatment, HCAECs showed mRNA expression of SST, SSTR-1, SSTR-2, and SSTR-5. The mRNA of SSTR-2 was expressed at a higher level than that of SSTR-1 and SSTR-5. However, SSTR-3 and SSTR-4 were not expressed or were minimally expressed. After treatment with TNF-α, the mRNA levels of SST, SSTR-1, SSTR-2, and SSTR-5 were significantly reduced in a dose-dependent fashion. TNF-α (1 ng/ml) reduced SST, SSTR-1, SSTR-2, and SSTR-5 by 93, 51, 85, and 99%, respectively, compared to controls (P < 0.001, t test). The immunoreactivity of SSTR-2 was also reduced after TNF-α treatment. SST-treated cells showed a significant reduction in [3H]thymidine incorporation in a dose-dependent manner. TNF-α treatment decreased SST inhibitory potential in cell proliferation.CONCLUSIONSHCAECs express SST, SSTR-1, SSTR-2, and SSTR-5, which are all decreased by TNF-α treatment. Furthermore, treatment with exogenous SST significantly reduces cell proliferation, and this inhibitory effect is also decreased by TNF-α.