Trailokyanath Panigrahi1, Shivapriya Shivakumar2, Rohit Shetty3, Sharon D'souza3, Everette Jacob Remington Nelson4, Swaminathan Sethu2, Nallathambi Jeyabalan5, Arkasubhra Ghosh6. 1. GROW Research Laboratory, Narayana Nethralaya Foundation, Bangalore, Karnataka, India; Department of Integrative Biology, School of Biosciences and Technology, VIT University Vellore, Tamil Nadu, 632014, India. 2. GROW Research Laboratory, Narayana Nethralaya Foundation, Bangalore, Karnataka, India. 3. Department of Cornea and Refractive Surgery, Narayana Nethralaya Eye Hospital, Narayana Health City, Bommasandra, Bangalore, Karnataka, India. 4. Department of Integrative Biology, School of Biosciences and Technology, VIT University Vellore, Tamil Nadu, 632014, India. 5. GROW Research Laboratory, Narayana Nethralaya Foundation, Bangalore, Karnataka, India. Electronic address: drnallathambi@narayananethralaya.com. 6. GROW Research Laboratory, Narayana Nethralaya Foundation, Bangalore, Karnataka, India. Electronic address: arkasubhra@narayananethralaya.com.
Abstract
PURPOSE: Cornea acts as a structural barrier and protects the eye from environmental stresses. Inflammation in ocular surface causes discomfort and visual distortion. Defective autophagy has been associated with inflammation and ocular surface diseases. Therefore, we explored the protective role of trehalose on inflammation and desiccation-triggered stress in human corneal cells in vitro and in dry eye patients. METHOD: TNF-α and desiccation stress induced human corneal cells (piHCF and HCE-T) with or without trehalose treatment were analyzed for the expression levels of inflammatory and autophagy related markers by qPCR, western blotting, multiplex ELISA and fluorescence imaging. Dry eye patients (N = 9) were enrolled and administered with trehalose in one eye and carboxymethylcellulose (CMC) in the contralateral eye (B.I.D, for 30 days). Dry eye signs OSDI, TBUT, Schirmer's Test, and tear cytokines were measured in dry eye patient's pre and post treatment. RESULTS: Cells treated with trehalose exhibits increased levels of autophagy markers LC3II and LAMP1 compared to untreated cells. Trehalose reduced the mRNA and secreted cytokines levels of IL-6, IL-8 and MCP-1 in corneal cells under TNF-α and desiccation stress mediated inflammation compared to controls. Further, trehalose reduced stress driven p38 phosphorylation in corneal cells. Additionally, topical administration of trehalose alleviated the clinical symptoms and tears cytokine levels in dry eye patients compared to CMC. CONCLUSION: Trehalose reduces stress induced inflammation through p38MAPK inhibition and autophagy activation. The anti-inflammatory mechanism of trehalose was independent to NFκB pathway. Further, topical administration of trehalose ameliorated dry eye associated symptoms and associated tear cytokines levels.
PURPOSE: Cornea acts as a structural barrier and protects the eye from environmental stresses. Inflammation in ocular surface causes discomfort and visual distortion. Defective autophagy has been associated with inflammation and ocular surface diseases. Therefore, we explored the protective role of trehalose on inflammation and desiccation-triggered stress in human corneal cells in vitro and in dry eyepatients. METHOD: TNF-α and desiccation stress induced human corneal cells (piHCF and HCE-T) with or without trehalose treatment were analyzed for the expression levels of inflammatory and autophagy related markers by qPCR, western blotting, multiplex ELISA and fluorescence imaging. Dry eyepatients (N = 9) were enrolled and administered with trehalose in one eye and carboxymethylcellulose (CMC) in the contralateral eye (B.I.D, for 30 days). Dry eye signs OSDI, TBUT, Schirmer's Test, and tear cytokines were measured in dry eyepatient's pre and post treatment. RESULTS: Cells treated with trehalose exhibits increased levels of autophagy markers LC3II and LAMP1 compared to untreated cells. Trehalose reduced the mRNA and secreted cytokines levels of IL-6, IL-8 and MCP-1 in corneal cells under TNF-α and desiccation stress mediated inflammation compared to controls. Further, trehalose reduced stress driven p38 phosphorylation in corneal cells. Additionally, topical administration of trehalose alleviated the clinical symptoms and tears cytokine levels in dry eyepatients compared to CMC. CONCLUSION:Trehalose reduces stress induced inflammation through p38MAPK inhibition and autophagy activation. The anti-inflammatory mechanism of trehalose was independent to NFκB pathway. Further, topical administration of trehalose ameliorated dry eye associated symptoms and associated tear cytokines levels.