Expression and Role of Nucleotide-Binding Oligomerization Domain 2 (NOD2) in the Ocular Surface of Murine Dry Eye.

Abstract

To investigate expression and role of nucleotide-binding oligomerization domain 2 (NOD2) in the ocular surface of experimental dry eye (EDE), which is a nod-like receptor member and is involved in innate immune response. C57/BL6 female mice were divided into the groups: untreated (UT), EDE, and NOD2 knockout (KO) mice exposed to desiccating stress for 14 days. Clinical parameters and levels of inflammatory cytokine were measured at 3,5,7, and 14 days. Immunofluorescent staining for NOD2 and Western blot for RIP2 and NF-κB were performed at 14 days. Flow cytometry, PAS staining and TUNEL staining were performed. After EDE induction, NOD2 was expressed in the corneal epithelium of the EDE group. The EDE group showed a significantly increased RIP2 expression compared to the UT and NOD2-KO groups. A significantly lower expression of NF-κB and lower levels of IL-1β, IL-6, IFN-γ, and TNF-α were noted in the NOD2-KO group than in the EDE group. The NOD2-KO group had lower CD11b+ and CD4+CCR5+ T cells, TUNEL-positive cells and corneal staining score and higher density of conjunctival goblet cell density, tear volume, and tear film break-up time than the EDE group. The UT group showed significant differences in inflammatory and clinical parameters compared to the EDE and NOD2-KO groups. The NOD2 receptor pathway induced inflammation and apoptosis by activation of RIP2 and NF-κB on the ocular surface of EDE, thereby reducing tear secretion. Therefore, NOD2 pathway may be involved in the pathogenesis of dry eye.