title: N-Acetylcysteine increases corneal endothelial cell survival in a mouse model of Fuchs endothelial corneal dystrophy.
abstract: The present study evaluated survival effects of N-acetylcysteine (NAC) on cultured corneal endothelial cells exposed to oxidative and endoplasmic reticulum (ER) stress and in a mouse model of early-onset Fuchs endothelial corneal dystrophy (FECD). Cultured bovine corneal endothelial cell viability against oxidative and ER stress was determined by CellTiter-Glo(®) luminescent reagent. Two-month-old homozygous knock-in Col8a2(L450W/L450W) mutant (L450W) and C57/Bl6 wild-type (WT) animals were divided into two groups of 15 mice. Group I received 7 mg/mL NAC in drinking water and Group II received control water for 7 months. Endothelial cell density and morphology were evaluated with confocal microscopy. Antioxidant gene (iNos) and ER stress/unfolded protein response gene (Grp78 and Chop) mRNA levels and protein expression were measured in corneal endothelium by real time PCR and Western blotting. Cell viability of H2O2 and thapsigargin exposed cells pre-treated with NAC was significantly increased compared to untreated controls (p < 0.01). Corneal endothelial cell density (CD) was higher (p = 0.001) and percent polymegathism was lower (p = 0.04) in NAC treated L450W mice than in untreated L450W mice. NAC treated L450W endothelium showed significant upregulation of iNos, whereas Grp78 and Chop were downregulated compared to untreated L450W endothelium by real time PCR and Western blotting. NAC increases survival in cultured corneal endothelial cells exposed against ER and oxidative stress. Systemic NAC ingestion increases corneal endothelial cell survival which is associated with increased antioxidant and decreased ER stress markers in a mouse model of early-onset FECD. Our study presents in vivo evidence of a novel potential medical treatment for FECD.
authors: Cornea & Anterior Segment Division, Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA; Department of Ophthalmology & Visual Science, College of Medicine, Catholic University of Korea, Seoul, Republic of Korea.
authors: Eun Chul
authors: EC
authors: Kim
authors: Cornea & Anterior Segment Division, Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
authors: Huan
authors: H
authors: Meng
authors: Cornea & Anterior Segment Division, Wilmer Eye Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA. Electronic address: aljun@jhmi.edu.
authors: Albert S
authors: AS
authors: Jun
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004195:Disease Models, Animal; D000091342:Endoplasmic Reticulum Chaperone BiP; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D016166:Free Radical Scavengers / Q000494:pharmacology*; D005642:Fuchs' Endothelial Dystrophy / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D006360:Heat-Shock Proteins / Q000235:genetics; D006861:Hydrogen Peroxide / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D018384:Oxidative Stress; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D019284:Thapsigargin / Q000633:toxicity; D051742:Transcription Factor CHOP / Q000235:genetics
chemical_list: C497250:Ddit3 protein, mouse; D000091342:Endoplasmic Reticulum Chaperone BiP; D016166:Free Radical Scavengers; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D012333:RNA, Messenger; D051742:Transcription Factor CHOP; D019284:Thapsigargin; D006861:Hydrogen Peroxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D000111:Acetylcysteine
keywords: ER stress; Fuchs endothelial corneal dystrophy (FECD); N-acetylcysteine (NAC); corneal endothelium; oxidative stress