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Abstract #86603 Published in IGR 21-2
Gene suppression of the ClC-2 chloride channel suppressed TGF-β1-induced proliferation, collagen synthesis, and collagen gel contraction mediated by conjunctival fibroblasts
Sun L; Cui R; Meng H; Liu X; Lu Y; Liu K; Jia L; Zheng YOphthalmic Research 2020; 0:
: BACKGROUND: ： Excessive scarring of filtering blebs is the main cause of surgical failure in glaucoma. Previous studies have highlighted the role of chloride channels in scar formation , whereas the role of chloride channels in scarring of filtering blebs has not been studied. OBJECTIVES: To investigate the effects of the ClC-2 chloride channel on scar formation of filtering blebs after glaucoma filtering surgery. METHODS: ClC-2 siRNA-transfected Human conjunctival fibroblasts (HconFs) were cultured in type I collagen gels in the presence of transforming growth factor (TGF)-β1. Collagen gel contraction was evaluated based on the gel area. 3D-cultured HConFs were treated with the chloride channel blocker NPPB in the presence of TGF-β1, and cell proliferation collagen synthesis and contractility were measured. The expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in HConFs were assessed by western blotting and q-PCR. RESULTS: TGF-β1induced cell proliferation, cell cycle progression, collagen synthesis, and collagen gel contraction in HConFs. TGF-β1 increased MMP-2 and MMP-9 levels but inhibited the expression of TIMPs. NPPB and ClC-2 siRNA transfection inhibited TGF-β2-induced cell proliferation, cell cycle progression, collagen synthesis, and collagen gel contraction, mediated by HConFs. TGF-β2-induced increases in MMP-2 and MMP-9 were also inhibited by NPPB and ClC-2 siRNA transfection, but TIMP expression was increased by NPPB and ClC-2 siRNA transfection. CONCLUSIONS: These findings demonstrate that ClC-2 chloride channels modulate TGF-β1-induced cell proliferation, collagen synthesis, and collagen gel contraction of HConFs by attenuating MMP-2 and MMP-9 production and by stimulating TIMP-1 production. NPPB may therefore prove to be of clinical value for the inhibition of scar formation of filtering blebs.
Full article
Classification:
3.8 Pharmacology (Part of: 3 Laboratory methods)
3.6 Cellular biology (Part of: 3 Laboratory methods)
3.4.2 Gene studies (Part of: 3 Laboratory methods > 3.4 Molecular genetics)
12.8.10 Woundhealing antifibrosis (Part of: 12 Surgical treatment > 12.8 Filtering surgery)
