Studing the signal transduction pathway of thymic stromal lymphopoietin in TGF-beta 1induced renal and lung cellular fibrosis

[[abstract]]糖尿病與肺炎是國人十大死因第四名與第六名，病程中分別會發展成腎纖維化與肺纖維化。許多研究指出生長因子間的表現失衡正是造成糖尿病腎纖維化與肺纖維化的主因。目前研究發現胸腺基質淋巴生成素（TSLP）與發炎疾病有著直接關聯。然而，沒有文獻指出TSLP所誘導的發炎反應是跟纖維化有關。因此，我們研究TSLP在NRK（腎臟纖維母細胞）與HFL-1（肺臟纖維母細胞）中的角色。然而，第一型乙型轉型生長因子（TGF-beta 1）是多功能性的細胞激素並且扮演引起腎纖維化與肺纖維化的重要角色。因此，本研究以第一型乙型轉型生長因子刺激腎臟纖維母細胞（NRK-49F）與肺臟纖維母細胞（HFL-1）進行體外培養，模擬腎細胞與肺細胞纖維化。接著，我們以酵素連結免疫吸附分析法與西方墨點法在TGF-beta 1（0.1、1或10 ng/mL）劑量和時間點下，發現NRK-49F 和HFL-1細胞外與細胞內中的TSLP隨著TGF-beta 1 劑量增加而上升。此外，加入外源性的TSLP（1、10 或100 ng/mL）劑量刺激下，會誘導NRK-49F 和HFL-1中纖維蛋白的增加。因此，我們以單獨（TGF-beta 1,TSLP）及協同的方式探討TGF-beta 1 與受器結合後Smad蛋白與其磷酸化程度的改變，發現在NRK-49F中不影響Smad蛋白（Smad2/3、pSmad2/3與Smad4）的表現，但會影響MAPK路徑（Raf-1、ERK-1與MEK-1）的表現。在HFL-1中，外源性的TSLP（1、10 或100 ng/mL）劑量刺激下，會影響Smad蛋白（Smad2/3、pSmad2/3與Smad4）的表現。另外，並探討施以抗發炎因子（如TSLP shRNA）對纖維化抑制的可行性。因此本研究主要探討TGF-beta 1 誘導的腎細胞纖維化與肺細胞纖維化的過程與TSLP的關係並且以TSLP shRNA抗纖維化作用機轉。 Diabetes and Pulmonary is the 4th and 6th leading causes of death in Taiwan. Many studies have demonstrated that dysregulation of growth factors might be the maincauses underlying the mechanism of diabetic renal fibrosis and Pulmonary fibrosis. The cytokine thymic stromal lymphopoietin (TSLP) has been linked to inflammatory diseases. None literature indicate TSLP induced the expression of inflammation response to correlate of fibrosis. Thus, we investigated the role of TSLP in NRK-49F cells (rat fibroblasts) and HFL-1 cells (lung fibroblasts). Transforming growth factor (TGF)-beta 1 is a multifunctional cytokine which also plays an important role in progressive renal and lung fibrosis. In this study, we used TGF-beta 1 induces renal and lung cellular fibrosis expression of TSLP. Moreover, exogenous TSLP significantly induce the expression of fibronectin in NRK-49F and HFL-1 cells. After inducing renal and lung cellular fibrosis by TGF-beta 1 and post-receptor signaling molecules (Smads and MAPK pathway) will be systemically investigated to elucidate the interactions between TSLP, renal and lung fibrosis. In addition, We show here TSLP expression was increased in the renal and lung fibroblast. Moreover, the feasibility using dual strategies by anti-inflammatory factor (i.e. TSLP shRNA) to control renal and lung fibrosis will also be examined in this project