Abstract
Objective: The objective of the study was to investigate the protective effects of Vitex negundo (VN) extracts against ultraviolet (UV) light C-induced oxidative stress in human corneal epithelial cells (HCEC) in vitro.
 Methods: The VN extracts were checked for their antioxidant activities by biochemical tests. The preliminary phytochemical analysis was done to check the presence of secondary metabolites. To trigger oxidative stress, the HCEC were exposed to UVC irradiation (200 J/m2). Then, different concentration of VN extracts was administered after 1 h of UVC irradiation. The effects of UVC irradiation and the extracts on the HCEC viability, morphology, cell migration by scratch assay, and reactive oxygen species (ROS) production by dichlorofluorescein diacetate method were measured.
 Results: VN ethanol (VNE) extract showed the highest antioxidant activity than the aqueous extract. Phytochemical screening shows the presence of alkaloids, carbohydrates, proteins, phenolics, and tannins. The extracts showed no cytotoxicity on the HCEC and also the VNE extract did not alter the morphology of the cells. Results suggest that the VNE extract enhanced the proliferation and migration of HCEC. The VNE extract significantly decreased UVC-induced oxidative toxicity by inhibiting the intracellular ROS production and maintained cell viability.
 Conclusion: VN extracts with its strong antioxidant potential protects HCEC from UVC-induced oxidative stress. The results suggest that these extracts can be used as prophylactic agents or as a therapeutic agent in overcoming UV light-induced damage in various ocular surface diseases such as pterygium and photokeratitis.
Highlights
Cornea acts as a primary and structural barrier of the eye
Our findings suggest that UVC radiation at a dose 200 J/m2 decreased the cell viability and induced production of reactive oxygen species (ROS) significantly which suggests that UV light especially UVC, can have a harmful impact on the corneal surface
Our results indicate that UVC radiation (254 nm) can induce oxidative damage to the corneal epithelium and that Vitex negundo (VN) extracts are effective in decreasing ROS production due to their high antioxidant activities
Summary
The corneal epithelium is frequently exposed to ultraviolet (UV) radiation such as UVA (315–400 nm), UVB (280–315 nm) from the sun, and UVC (200–280 nm) through occupational exposures [1]. The cornea is known to absorb 100% of UVC without allowing them to penetrate further into the interior of the eye [2]. Imbalance in the antioxidant defense mechanism takes place during UV exposure leading to injury in the cornea and other parts of the eye [3]. Excessive exposure to these radiations leads to oxidative stress in the ocular surface cells resulting in epithelial exfoliation followed by extreme pain, a condition known as photokeratitis [4]
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