Inhibition of Pirfenidone on TGF-beta2 Induced Proliferation, Migration and Epithlial-Mesenchymal Transition of Human Lens Epithelial Cells Line SRA01/04

• DOI: 10.1371/journal.pone.0056837
• Publication Date: 2013-02-21T23:49:18Z
• Authors: Yangfan Yang; Yiming Ye; Xianchai Lin; Kaili Wu; Minbin Yu
• Keywords: Pirfenidone; Viability assay; MTT assay; Trypan blue

Background Posterior capsular opacification (PCO) is a common complication of cataract surgery. Transforming growth factor-β2 (TGF-β2) plays important roles in the development PCO. The existing pharmacological treatments are not satisfactory and can have toxic side effects. Methodologies/Principal Findings We evaluated effect pirfenidone on proliferation, migration epithlial-mesenchymal transition human lens epithelial cell line SRA01/04 (HLECs) vitro. After treatment with 0, 0.25, 0.5 mg/ml pirfenidone, proliferation was measured by MTT assay. Cell viability determined trypan blue exclusion assay measurement lactate dehydrogenase (LDH) activity released from damaged cells. And scratch absence or presence transforming (TGF-β2). expressions TGF-β2 SMADs were real-time RT-PCR, western blot, immunofluorescence analyses. mesenchymal phenotypic marker fibronectin (FN) demonstrated Immunocytofluorescence cells had high viability, which did vary across different concentrations (0 [control] 0.3, 1.0 mg/ml) after 24 hours. Pirfenidone (0∼0.5 no significant cytotoxicity LDH significantly inhibited TGF-β2-induced effects dose-dependent, fibroblastic phenotypes expression FN showed dose-dependent decreases mRNA protein levels SMADs. also depressed blocked nuclear translocation Conclusion inhibits at nontoxic concentrations. This may be achieved down regulation TGF-β/SAMD signaling