Hepatocyte Growth Factor Suppresses Transforming Growth Factor-Beta-1 and Type Ⅲ Collagen in Human Primary Renal Fibroblasts

肝細胞生長因子抑制人腎間質成纖維細胞膠原合成和轉化生長因子－β1表達

牟姍(Shan Mou) ； 王琴(Qin Wang,) ； 施蓓莉(Bei-Li Shi) ； 顧樂怡(Le-Yi Gu) ； 倪兆慧(Zhao-Hui Ni)

《The Kaohsiung Journal of Medical Sciences》 25卷11期 (2009/11) Pp. 577-587

摘要

糖尿病腎病過程中，當腎小球濾過率下降，常常伴有腎小管間質病變。此項研究中，採用人腎臟成纖維細胞培養系統，探討肝細胞生長因子(HGF)對高糖誘導的腎間質成纖維細胞損傷產生的作用。用消化法體外培養人腎間質成纖維細胞，用D-葡萄糖刺激細胞，造成高糖環境。另外高糖環境下外源加入rHGF誘導細胞。以酶聯免疫吸附法檢測III型膠原情況，用RT-PCR檢測細胞HGF和TGF-β1基因表達，Westernblot方法檢測TGF-β1蛋白合成。高糖預處理過的細胞III型膠原合成增加。同時觀察到高糖作用早期HGF表達升高，隨著作用時間延長，其表達量下降，而TGF-β1出現持續高表達。同時外源加入HGF可以抑制TGF-β1基因和蛋白表達，對III型膠原合成產生抑制作用。體外高糖環境促進III型膠原合成和TGF-β1持續高表達。rHGF可以部分抑制III型膠原和TGF-β1表達，從而發揮延緩間質損傷的作用。局部的HGF/c-met系統在糖尿病腎病過程中起重要作用。

關鍵字

肝細胞生長因子受體 ； 糖尿病腎病 ； 肝細胞生長因子 ； 間質纖維化 ； 轉化生長因子－β1

並列摘要

Tubulointerstitial changes in the diabetic kidney correlate closely with renal fibrosis, and transforming growth factor-beta-1 (TGF-β1) is thought to play a key role in this process. In contrast, hepatocyte growth factor (HGF) has shown therapeutic effects on injured renal tubules in animal models. This study was undertaken to test the hypothesis that the preventive effects of HGF may result from interventions in TGF-β1-mediated signaling and collagen III secretion. We examined the expression of HGF/HGF receptor (c-Met) and TGF-β1 in renal fibroblasts at multiple time points. The effects of recombinant human HGF on TGF-β1 expression were studied by RT-PCR and Western blotting, and the levels of collagen III were measured by ELISA. In the high-glucose condition, the expression of HGF and c-Met in renal fibroblasts was detected as early as 6 hours following cell culture while the level of TGF-β1 peaked at 96 hours. The addition of recombinant human HGF to the culture media dose-dependently inhibited TGF-β1 mRNA expression and reduced collagen III secretion by 34%. These results indicate that, during hyperglycemia, HGF inhibits TGF-β1 signaling and type III collagen activation in interstitial fibroblasts. Furthermore, we should recognize that changes in the balance between HGF and TGF-β1 might be decisive in the pathogenesis of chronic renal fibrosis. Therefore, administration of HGF to restore this balance may offer a novel therapeutic intervention in managing renal fibrogenesis in diabetic nephropathy.

並列關鍵字

c-Met ； diabetic nephropathy ； hepatocyte growth factor ； interstitial fibrosis ； transforming growth factor-β1