Thrombin刺激肺部纖維母細胞產生結締組織生長因子經JAK2及STAT3之訊息角色探討

The Roles of JAK2 and STAT3 in Thrombin-Induced Connective Tissue Growth Factor Expression in Human Pulmonary Fibroblasts

白惠珍(Hui-ChenPai)

指導教授 ： 林建煌

共同指導教授 ： 陳炳常

臺北醫學大學/醫學院/醫學科學研究所/碩士(2007年)

https://doi.org/10.6831/TMU.2007.00043

摘要

在本篇論文中，我們將探討thrombin調控纖維母細胞connective tissue growth factor (CTGF)的表現之詳細分子機轉。在WI-38纖維母細胞中，thrombin可誘導CTGF蛋白表現及CTGF-luciferase的活性。給予AG490 (JAK抑制劑)及JAK DN可依劑量反應關係，抑制thrombin誘導CTGF蛋白的表現及CTGF-luciferase的活性。 Thrombin以時間相關性地誘導JAK2之 Tyr1007/1008 的磷酸化。細胞給予轉染STAT3 DN及 STAT ODN (oligodeoxynucleotides)抑制thrombin誘導CTGF蛋白的表現及CTGF-luciferase的活性。 Thrombin誘導STAT 之Tyr705的磷酸化呈現時間相關性，並且藉由DNA-binding affinity pull down assay證實thrombin可以誘導STAT結合至CTGF基因起始區上。Thrombin誘導STAT3的磷酸化可被AG490及JAK DN所抑制。我們也發現thrombin誘導CTGF蛋白表現也可被兩種STAT3結合序列的ODN (STAT3-1 ODN及STAT3-2 ODN)所抑制。再者，thrombin所誘導CTGF蛋白表現及CTGF-luciferase的活性，可被c-Src DN抑制。Thrombin誘導STAT3磷酸化及JAK2磷酸化也可被c-Src DN抑制。綜合以上的結果推測thrombin可經由c-Src，JAK2及STAT3訊息路徑來誘導WI-38細胞之CTGF蛋白的表現。

關鍵字

肺纖維化 ； 結締組織生長因子

並列摘要

In this study, we investigated the signaling pathway by thrombin- induced CTGF expression. Thrombin induced increase in CTGF protein expression and CTGF-luciferase activity in WI-38 lung fibroblasts. AG490 (JAK inhibitor) and dominant negative of JAK2 (JAK2 DN) inhibited thrombin-CTGF expression and CTGF-luciferase activity in a dose-dependent manner. Thrombin also caused a time-dependent increase JAK2 phosphorylation at Tyr1007/1008. Transfection of cells with STAT3 DN and STAT3 ODN (oligodeoxynucleotides) inhibited CTGF protein expression and CTGF-luciferase activity. Thrombin induced STAT3 at Tyr705 phosphorylation in a time-dependent manner, which was inhibited by AG490 and JAK2 DN. Thrombin induced STAT binding to CTGF promoter by DNA-binding affinity pull down assay. We also found that thrombin-induced CTGF protein expression was inhibited by STAT3-1 oligodeoxynucleotides(ODN) and STAT3-2 (ODN). Furthermore, thrombin-indused CTGF protein expression and CTGF-luciferase activity was inhibited by cell transfected with c-Src DN. Thrombin induced STAT3 phosphorylation and JAK2 phosphorylation also inhibited by c-Src DN. Taken together, thrombin though Src, JAK2, and STAT3 signaling pathway to induced CTGF protein expression and activity in lung fibroblasts.

並列關鍵字

lung fibroblasts ； CTGF ； JAK2 ； STAT3

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