Abstract
BackgroundPosterior capsular opacification (PCO) is a common complication of cataract surgery. Transforming growth factor-β2 (TGF-β2) plays important roles in the development of PCO. The existing pharmacological treatments are not satisfactory and can have toxic side effects.Methodologies/Principal FindingsWe evaluated the effect of pirfenidone on proliferation, migration and epithlial-mesenchymal transition of human lens epithelial cell line SRA01/04 (HLECs) in vitro. After treatment with 0, 0.25, and 0.5 mg/ml pirfenidone, cell proliferation was measured by MTT assay. Cell viability was determined by trypan blue exclusion assay and measurement of lactate dehydrogenase (LDH) activity released from the damaged cells. And cell migration was measured by scratch assay in the absence or presence of transforming growth factor-β2 (TGF-β2). The expressions of TGF-β2 and SMADs were evaluated with real-time RT-PCR, western blot, and immunofluorescence analyses. The mesenchymal phenotypic marker fibronectin (FN) was demonstrated by Immunocytofluorescence analyses. The cells had high viability, which did not vary across different concentrations of pirfenidone (0 [control] 0.3, 0.5 or 1.0 mg/ml) after 24 hours. Pirfenidone (0∼0.5 mg/ml) had no significant cytotoxicity effect on SRA01/04 by LDH assay. Pirfenidone significantly inhibited the proliferation and TGF-β2-induced cell migration and the effects were dose-dependent, and inhibited TGF-β2-induced fibroblastic phenotypes and TGF-β2-induced expression of FN in SRA01/04 cells. The cells showed dose-dependent decreases in mRNA and protein levels of TGF-β2 and SMADs. Pirfenidone also depressed the TGF-β2-induced expression of SMADs and blocked the nuclear translocation of SMADs in cells.ConclusionPirfenidone inhibits TGF-β2-induced proliferation, migration and epithlial-mesenchymal transition of human lens epithelial cells line SRA01/04 at nontoxic concentrations. This effect may be achieved by down regulation of TGF-β/SAMD signaling in SRA01/04 cells.
Highlights
Posterior capsular opacification (PCO) is the most frequent complication of cataract surgery and mainly caused by fibrosis on lens capsular bag and/or posterior capsule [1]
Previous studies have shown that many growth factors, especially transforming growth factor-b2 (TGF-b2), play important roles in the development of PCO, which may trigger a series of events during wound healing process [2,3,4,5]
Considering the important effect of TGF-b2 signaling pathway, we examined the roles of PFD on mRNA or protein expression TGF-b2 and SMADs and the nuclear translocation of SMADs
Summary
Posterior capsular opacification (PCO) is the most frequent complication of cataract surgery and mainly caused by fibrosis on lens capsular bag and/or posterior capsule [1]. Previous studies have shown that many growth factors, especially transforming growth factor-b2 (TGF-b2), play important roles in the development of PCO, which may trigger a series of events (including cell proliferation, differentiation, migration and capsular fibrosis) during wound healing process [2,3,4,5]. Methodologies/Principal Findings: We evaluated the effect of pirfenidone on proliferation, migration and epithlialmesenchymal transition of human lens epithelial cell line SRA01/04 (HLECs) in vitro. After treatment with 0, 0.25, and 0.5 mg/ml pirfenidone, cell proliferation was measured by MTT assay. Cell migration was measured by scratch assay in the absence or presence of transforming growth factor-b2 (TGF-b2). Pirfenidone significantly inhibited the proliferation and TGF-b2-induced cell migration and the effects were dose-dependent, and inhibited TGF-b2induced fibroblastic phenotypes and TGF-b2-induced expression of FN in SRA01/04 cells. Pirfenidone depressed the TGF-b2-induced expression of SMADs and blocked the nuclear translocation of SMADs in cells
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