In Vitro Expression Analysis of Cytokines and ROS-Related Genes in Human Corneal Fibroblasts and Keratocytes of Healthy and Keratoconus Corneas

Abstract

ABSTRACT Purpose To investigate expression of cytokines and ROS-related genes in stromal cells of healthy and keratoconus (KC) corneas. Methods Expression analysis was performed for cytokines including several interleukins (IL), Tumor necrosis factor-α (TNF-α), Transforming growth factor-β1 (TGF-β1), Interferon-γ (IFN-γ) and ROS-related genes such as Catalase, Glutathione peroxidase 1, NADPH oxidase 1, superoxide dismutase 1 in corneal fibroblasts (HCFs/KC-HCFs) or keratocytes (Keratocytes/KC-Keratocytes) by qPCR and ELISA. Results Gene and protein expression of most inflammatory markers was decreased in keratocytes compared to fibroblasts, whereas no differences were found between healthy and keratoconus cells for the majority of cytokines measured. TNF-α expression was increased at gene (KC keratocytes) and protein levels (supernatant of Keratocytes/KC-Keratocytes) compared to corneal fibroblasts. No differential expression of ROS-related genes was detected between healthy and diseased cells in both fibroblasts and keratocytes. Conclusion Increased expression of several inflammatory markers described as altered in KC was not evident in KC cells in vitro.