Kang Jeong-Han, Cho Hyun-Ji, Lee In-Seon, Kim Moonkyu, Lee In-Kyu, Chang Young-Chae
Department of Pathology, Research Institute of Biomedical Engineering, Catholic University of Daegu School of Medicine, Daegu 705-034, Korea.
Proteomics. 2009 Oct;9(19):4445-56. doi: 10.1002/pmic.200800941.
Transforming growth factor-beta1 (TGF-beta1) has a wide range of biological functions such as the regulation of cell growth, differentiation, and immunological response in various types of cells. Particularly, TGF-beta1 induces plasminogen activator inhibitor-1 (PAI-1) as a major target protein. PAI-1 is associated with fibrosis, thrombosis, and metabolic disorders. In this study, to identify proteins potentially involved in TGF-beta1-induced fibrosis processes, we performed a proteomic analysis of TGF-beta1-induced normal rat kidney cells exposed to ascofuranone (AF). In these cells, we detected 1500 proteins, with 74 differentially expressed proteins identified by MALDI-TOF and reference to the NCBI and Swiss-Prot databases, including PAI-1, peroxisome prdifesator-activated receptor, prohibitin, glutamate formyltransferase, LIM domain protein 1, LASP-1, porphobilinogen deaminase, and peroxiredoxin 2. We also found that AF suppresses expression of profibrotic factors induced by TGF-beta in renal fibroblasts, including matrix proteins and PAI-1. AF was also shown to inhibit selectively phosphorylation of epidermal growth factor receptor, and downstream kinases such as extracellular signal-regulated kinase 1/2 (ERK-1/2). Further ongoing analysis of fibrosis-related proteins will determine AF's potential for application in fibrotic diseases and therapeutics.
转化生长因子-β1（TGF-β1）具有广泛的生物学功能，如调节各类细胞的生长、分化及免疫反应。特别地，TGF-β1诱导纤溶酶原激活物抑制剂-1（PAI-1）作为主要靶蛋白。PAI-1与纤维化、血栓形成及代谢紊乱相关。在本研究中，为鉴定可能参与TGF-β1诱导的纤维化过程的蛋白质，我们对暴露于当归呋内酯（AF）的TGF-β1诱导的正常大鼠肾细胞进行了蛋白质组学分析。在这些细胞中，我们检测到1500种蛋白质，通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）并参考美国国立生物技术信息中心（NCBI）和瑞士生物信息学研究所（Swiss-Prot）数据库鉴定出74种差异表达蛋白质，包括PAI-1、过氧化物酶体增殖物激活受体、抑制素、谷氨酸甲酰基转移酶、LIM结构域蛋白1、LASP-1、胆色素原脱氨酶和过氧化物还原酶2。我们还发现AF抑制肾成纤维细胞中TGF-β诱导的促纤维化因子的表达，包括基质蛋白和PAI-1。AF还显示出选择性抑制表皮生长因子受体及其下游激酶如细胞外信号调节激酶1/2（ERK-1/2）的磷酸化。对纤维化相关蛋白质的进一步分析将确定AF在纤维化疾病及治疗中的应用潜力。
