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Jin Hyoung Park, Soon-Suk Kang, Hungwon Tchah; Nerve Growth Factor Attenuate Apoptosis and Inflammation in Diabetic Cornea. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5390.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the effects of nerve growth factor (NGF) on apoptosis and inflammation in diabetic cornea using human corneal epithelial cells (HCECs) and streptozotocin-induced diabetic rat cornea.
The effects of NGF on high glucose-induced apoptosis were investigated in HCECs using Annexin-V and PI staining, and cleaved caspase-3 and BAX expression levels using immunoblotting. The effects of NGF on inflammatory response were examined using quantified IL-1β and TNF-a expression levels by multiplex cytokine analysis, and NF-kB activation and IkB-a degradation by western blot analysis. Diabetes was induced by intraperitoneal streptozotocin injection in male Sprague–Dawley rats. The SD rats were divided into 3 groups. (1) Control group (2) Diabetic control group (3) NGF group; Topical NGF were given 3 times a day for 3 weeks. Apoptosis was detected by TUNEL assay. The expressions of caspase-3 and IL-1 β were studied by immunohistochemistry in corneal tissues.
High glucose at 25 mM stimulated ROS generation, apoptosis, and release of inflammatory cytokines in HCECs. The addition of NGF markedly reduced the high glucose-induced ROS activation, Annexin-PI-positive cells, and levels of cleaved caspase-3, BAX, IL-1β, and TNF-α in HCECs. Enhanced apoptotic and inflammatory responses and expressions of cleaved caspase-3 and IL-1 β in diabetic corneas of rat were observed. These responses were markedly reduced by NGF.
Apoptosis and inflammation were enhanced in diabetic cornea and NGF contributed to reduction of apoptotic and inflammatory response in vivo and in vitro conditions. NGF may be a novel therapeutic approach for managing diabetic keratopathy.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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