Abstract

As the prevalence of microbial keratitis increases, it creates an environment conducive to genotoxicity response. A potential connection between growth arrest and DNA-damage-inducible 45 gamma (GADD45G) gene expression has not been proven in the corneal epithelial cells. The aim of this study was to determine whether lipopolysaccharide (LPS) enhances genotoxicity, DNA damage, and inflammatory responses in human corneal epithelial cells (HCECs) in vitro. In a set of parameters, cytotoxicity, reactive oxygen species, mitochondrial membrane potential, DNA damage, inflammatory response, and apoptosis were assessed. LPS (1, 5, and 10 μg/mL) treated HCECs were increased reactive oxygen species formation, mitochondrial membrane depolarization, and genotoxicity in a concentration-dependent manner. Similarly, NF-κB, PARP1, and TP53 were also overexpressed in the LPS treated HCECs. 24 hours after LPS induction, micronucleus scoring, and proapoptotic factors were also increased. Among them, the GADD45G, NF-κB, and γH2AX were overexpressed both on the mRNA and protein levels in LPS (10 μg/mL) treated HCECs. In our study, we show that the GADD45G signaling can trigger genotoxic instability in HCECs exposed to LPS. Therefore, understanding the factors contributing to infectious keratitis, such as GADD45G, NF-κB, and γH2AX signaling, may help to develop antigenotoxic and anti-inflammatory therapies for corneal dystrophy and epithelial cell remodeling.

Highlights

  • Corneoscleral interface of the limbal epithelium serves as a barrier that protects the eye from bacterial invasion and serves as a source of stem cells [1]

  • Detection of Cell Viability in human corneal epithelial cells (HCECs) Treated with LPS

  • The viability of HCECs treated with LPS was evaluated at 24, 48, and 72 hours (Figure 1)

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Summary

Introduction

Corneoscleral interface of the limbal epithelium serves as a barrier that protects the eye from bacterial invasion and serves as a source of stem cells [1]. The cornea is affected by a microbial infection that manifests as corneal inflammations, corneal thinning, and corneal perforation and scarring [2, 3]. It is characterized by the presence of diffuse or localized infiltrates on the anterior surface of the ocular tissues and corneal epithelium. There is an increasing incidence of microbial infectious diseases; contact lens wearers make up one of the biggest risk factors for causing infectious keratitis [6]. Numerous epidemiological studies have consistently indicated that overnight contact lens wear increases the incidence of microbial keratitis [7,8,9]

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