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    題名: Shc在thrombin誘導肺部上皮細胞IL-8/CXCL8表現所扮演的角色
    The Role of Shc on Thrombin-Induced IL-8/CXCL8 Expression in Lung Epithelial Cells
    作者: 江佳潔
    Chia Chieh Chiang
    貢獻者: 醫學科學研究所
    關鍵詞: 肺部上皮細胞
    IL-8/CXCL8
    凝血酵素
    Shc
    IL-8/CXCL8
    thrombin
    lung epthelial cell
    A549
    日期: 2009
    上傳時間: 2009-09-14 15:31:10 (UTC+8)
    摘要: Thrombin除了在形成血栓及維持血液恆定扮演重要的角色外,也是調控肺部發炎反應的重要因子。本實驗室之前發表的論文中指出,在人類肺部上皮細胞中,c-Src可以媒介thrombin誘導NF-κB 的活性及IL-8/CXCL8的表現。在本篇研究中將探討在人類肺部上皮細胞中,c-Src依賴Shc/Raf-1/ERK訊息傳遞路徑在thrombin誘導IL-8/CXCL8的表現所扮演的角色。Thrombin所誘導IL-8/CXCL8的釋放及κB-luciferase的活性可被Shc siRNA所抑制。A549細胞給予thrombin刺激,可依時間依賴的方式誘導Shc之Tyr-239/240及Tyr-317的磷酸化。然而只有Shc之Tyr-239/240位置的磷酸化可被c-Src DN所抑制。相同地,thrombin也可依時間依賴的方式誘導Raf-1及ERK的磷酸化。Thrombin所誘導IKKα/β的磷酸化、κB-luciferase的活性及IL-8/CXCL8的釋放可被GW 5074 (Raf-1的抑制劑) 及PD 98059 (MEK的抑制劑) 所抑制。更進一步的研究顯示, thrombin可依時間相關性誘導PAR 1、 Gβ、c-Src及Shc蛋白的交互作用。綜合以上的實驗結果,可推測出在肺部上皮細胞中,thrombin可經由PAR1/Gβ/c-Src依賴Shc/Raf-1/ERK/訊息傳遞路徑來增加IKKα/β/NF-κB的活性,進而調控IL-8/CXCL8的表現及釋放。

    Thrombin, excluding the functions in thrombosis and hemostasis, also plays an important role in the lung inflammation. Our previously report showed that c-Src mediates thrombin-induced NF-κB activation and IL-8/CXCL8 expression in lung epithelial cells. This study investigated the mechanism of the c-Src-dependent Shc/Raf-1/ERK signal pathway involved in thrombin-induced IL-8/CXCL8 release in human lung epithelial cells. Thrombin-induced IL-8/CXCL8 expression and κB-luciferase activity were inhibited by Shc siRNA. Treatment of A549 cells with thrombin caused Shc phosphorylation at Tyr-239/240 and Tyr-317 in a time-dependent manner. Transfected c-Src DN inhibited thrombin-induced Shc phosphorylation at Tyr-239/240, but not at Tyr-317. Thrombin also induced Raf-1 and ERK phosphorylation in a time-dependent manner. Pretreatment of A549 cells with GW 5074 (a Raf-1 inhibitor) and PD 98059 (a MEK inhibitor) both inhibited thrombin-induced IKKα/β phosphorylation, κB-luciferase activity, and IL-8/CXCL8 release. Furthermore, studies revealed that thrombin enhances the recruitment of PAR1, Gβ, Shc to c-Src in a time-dependent manner. Take together, these results indicated that thrombin activates the the PAR1/Gβ/c-Src-dependent Shc/Raf-1/ERK signaling pathway to induce IKKα/β/NF-κB activation, which in turn induces IL-8/CXCL8 expression and release in human lung epithelial cells.
    資料類型: thesis
    顯示於類別:[醫學科學研究所] 博碩士論文

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