Int J Med Sci 2020; 17(6):734-744. doi:10.7150/ijms.43238

Research Paper

The in vitro anti-fibrotic effect of Pirfenidone on human pterygium fibroblasts is associated with down-regulation of autocrine TGF-β and MMP-1

Yijin Tao1*, Qin Chen2*, Can Zhao3, Xiao Yang1, Qing Cun1, Wenyan Yang1, Yuan Zhang4, Yingting Zhu4✉, Hua Zhong1✉

1. Department of Ophthalmology, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, China.
2. Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 211166, China.
3. Shandong Eye Hospital, State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences.
4. Tissue Tech, Inc., Miami, FL, 33126, USA.
*These authors contributed equally to this work.

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Citation:
Tao Y, Chen Q, Zhao C, Yang X, Cun Q, Yang W, Zhang Y, Zhu Y, Zhong H. The in vitro anti-fibrotic effect of Pirfenidone on human pterygium fibroblasts is associated with down-regulation of autocrine TGF-β and MMP-1. Int J Med Sci 2020; 17(6):734-744. doi:10.7150/ijms.43238. Available from http://www.medsci.org/v17p0734.htm

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Abstract

We aimed to investigate the in vitro effect of pirfenidone (PFD) on proliferation, migration and collagen contraction of human pterygium fibroblasts (HPFs). HPFs were obtained from tissue explants during pterygium surgery. After treatment with pirfenidone, the HPFs proliferation was measured by MTT, cell cycle progression measured by flow cytometry, cell migration measured by the scratch assay, and cell contractility evaluated in fibroblast-populated collagen gels. The expression of TGF-β1, TGF-β2, MMP-1 and TIMP-1 were also determined with quantitative PCR, western blot and immunofluorescence staining. Results showed pirfenidone markedly inhibited HPFs proliferation with an IC50 of approximately 0.2 mg/ml. After treatment with 0.2 mg/ml pirfenidone for 24 hours, HPFs were at G0/G1 cell cycle arrest, with significantly reduced cell migration capability and collagen contraction, decreased mRNA and protein expressions of TGF-β1, TGF-β2 and MMP-1, and no alterations of TIMP-1 expression. Thus, we have concluded that pirfenidone at 0.2 mg/ml inhibits proliferation, migration, and collagen contraction of HPFs, which is associated with decreased expression of TGF-β and MMP-1, and pirfenidone might represent a potentially therapeutic agent to prevent the recurrence of pterygium after surgery.

Keywords: pirfenidone, human pterygium fibroblasts, proliferation, migration, collagen contraction, TGF-β, MMP