Liu Jingjing, Song Cheng, Xiao Qiming, Hu Gaoyun, Tao Lijian, Meng Jie
*Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha; †Department of Respiratory Medicine, Central Hospital of Wuhan, Tongji Medical College Huazhong University of Science and Technology, Wuhan; ‡Pharmaceutical School, Central South University; and §Department of Nephrology Medicine, Xiangya Hospital, Central South University, Changsha; and ∥State Key Lab of Respiratory Disease and National Clinical Research Center for Respiratory Disease, Guangzhou, China.
Shock. 2015 Feb;43(2):201-7. doi: 10.1097/SHK.0000000000000273.
Caveolin-1 plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. We previously showed that fluorofenidone (FD), a novel pyridine agent, can attenuate bleomycin-induced experimental pulmonary fibrosis and restore the production of caveolin-1. In this study, we explore mainly whether caveolin-1 plays a critical role in the anti-pulmonary fibrosis effects of FD in vitro. The normal human lung fibroblasts (NHLFs) were cultured with transforming growth factor-β1 (TGF-β1) and then were treated with FD. Subsequently, NHLFs transfected with cav-1-siRNA were treated with TGF-β1 and/or FD. The expressions of α-smooth muscle actin (α-SMA), fibronectin, collagen I, caveolin-1, phosphorylated extracellular signal-regulated kinase (p-ERK), phosphorylated c-Jun N-terminal kinase (p-JNK), and phosphorylated P38 were measured by Western blot and/or real-time polymerase chain reaction. Fluorofenidone attenuated TGF-β1-induced expressions of α-SMA, fibronectin, and collagen I; inhibited phosphorylation of ERK, JNK, and P38; and restored caveolin-1 protein expression but cannot increase caveolin-1 mRNA level in vitro. After caveolin-1 was silenced, FD could not downregulate TGF-β1-induced expressions of α-SMA, fibronectin, and collagen I or phosphorylation of ERK, JNK, and P38. These studies demonstrate that FD, a potential antifibrotic agent, may attenuate TGF-β1-induced activation of NHLFs by restoring the expression of caveolin-1.
小窝蛋白-1在特发性肺纤维化的发病机制中起重要作用。我们之前表明，新型吡啶类药物氟非尼酮（FD）可减轻博来霉素诱导的实验性肺纤维化，并恢复小窝蛋白-1的产生。在本研究中，我们主要探讨小窝蛋白-1在FD体外抗肺纤维化作用中是否起关键作用。将正常人肺成纤维细胞（NHLFs）用转化生长因子-β1（TGF-β1）培养，然后用FD处理。随后，用cav-1-siRNA转染的NHLFs用TGF-β1和/或FD处理。通过蛋白质免疫印迹法和/或实时聚合酶链反应检测α-平滑肌肌动蛋白（α-SMA）、纤连蛋白、I型胶原、小窝蛋白-1、磷酸化细胞外信号调节激酶（p-ERK）、磷酸化c-Jun氨基末端激酶（p-JNK）和磷酸化P38的表达。氟非尼酮减轻了TGF-β1诱导的α-SMA、纤连蛋白和I型胶原的表达；抑制了ERK、JNK和P38的磷酸化；并恢复了小窝蛋白-1蛋白表达，但在体外不能增加小窝蛋白-1 mRNA水平。小窝蛋白-1沉默后，FD不能下调TGF-β1诱导的α-SMA、纤连蛋白和I型胶原的表达或ERK、JNK和P38的磷酸化。这些研究表明，潜在的抗纤维化药物FD可能通过恢复小窝蛋白-1的表达来减轻TGF-β1诱导的NHLFs活化。
