Effects of Acetazolamide on Collagen Metabolism in Keratoconus Fibroblasts

Abstract

Keratoconus is characterized by the progressive thinning and protrusion of central cornea. Dysregulated collagen metabolism is one of the causes of this disease. In this study, the effect of acetazolamide on collagen synthesis in keratoconus fibroblasts and its mechanism were explored, which could provide a reference for the clinical treatment of keratoconus. The cultured keratoconus fibroblasts were treated with 100 μmol/L acetazolamide solution. The expression of collagen metabolism-related genes was then analyzed by qPCR. The collagen content was detected by ELISA and Western Blot. The intracellular ROS content was analyzed by fluorescent probe DCFH-DA. The changes of MDA content and the total antioxidant capacity were detected by spectrophotometry. It was suggested that after acetamide treatment, the expression of COLs and LOXs, of which coded enzymes paticipated in collagen synthesis and crosslinking, were significantly up-regulated, while the expression of collagen degradation genes including MMP2, MMP3, and MMP9 were significantly down-regulated. The contents of type I collagen and total collagens were both reduced by acetamide treatment. In addition, the total antioxidant capacity of cells and the expression of the antioxidant enzyme gene NQO-1 were significantly up-regulated, while the contents of ROS and MDA were significantly decreased. The results indicated that acetazolamide could induce the synthesis and inhibit the degradation of collagens in keratoconus fibroblasts by reducing the intracellular oxidative stress levels.

Keywords

• acetazolamide
• keratoconus
• collagen
• oxidative stress