The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)
The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)
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DOI：
10.1007/s00417-008-0990-5
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发表时间：
2009-01
期刊：
影响因子：
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通讯作者：
Kazuhiko Watanabe;Xue-Yun Zhang;K. Kitagawa;Tatsuya Yunoki;A. Hayashi
中科院分区：
文献类型：
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作者：
Kazuhiko Watanabe;Xue-Yun Zhang;K. Kitagawa;Tatsuya Yunoki;A. Hayashi
本研究旨在探讨α 2肾上腺素能受体（α2-ADR）激动剂可乐定对白细胞介素1β（IL-1β）刺激的人视网膜色素上皮细胞株（ARPE-19）血管内皮生长因子（VEGF）表达和分泌的影响。半定量逆转录聚合酶链反应（RT-PCR）检测ARPE-19细胞α 2 A、α 2 B和α 2 C mRNA的表达。在IL-1β刺激前，向无血清培养基中加入可乐定和参与细胞内信号转导调节的蛋白激酶抑制剂。α2-ADR拮抗剂育亨宾在可乐定加入前30 min负荷。结果RPE细胞表达α 2 A-ADR、α 2 B-ADR和α 2 C-ADR mRNA; p38 MAPK和MEK 1/2抑制剂可乐定可抑制IL-1β刺激的RPE细胞VEGF蛋白和mRNA的表达。结论可乐定可能通过抑制IL-1 β激活的p38 MAPK和MEK 1/2信号转导途径抑制VEGF的表达。
BackgroundThe purpose of this study was to investigate the effect of clonidine, an alpha2-adrenergic receptor (α2-ADR) agonist, on vascular endothelial growth factor (VEGF) expression and secretion in the human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β).MethodsAlpha2-ADRs (α2A, α2B, and α2C) mRNA expression in ARPE-19 cells was examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Clonidine and inhibitors against protein kinases that are involved in the regulation of the intracellular signal transduction were added to serum-free medium before stimulation of IL-1β. The α2-ADR antagonist, Yohimbine, was loaded 30 min before the addition of clonidine. The expression of VEGF mRNA and protein was measured by real-time PCR and enzyme-linked immunosorbent assay.ResultsAlpha2A-ADR, α2B-ADR, and α2C-ADR mRNA was expressed in RPE cells. Clonidine, an inhibitor of p38MAPK and MEK1/2, inhibited the expression of VEGF protein and mRNA in the RPE cells stimulated with IL-1β. The inhibitory effect of clonidine on the secretion of VEGF protein stimulated with IL-1β was blocked by α2-ADR antagonists.ConclusionsThe effect of clonidine on the expression of VEGF may be via suppression of the p38MAPK and MEK1/2 signal transduction pathways activated with IL-1β.