Abstract

Dry Eye Disease (DED) is a multifactorial ocular surface disorder characterized by tear-film instability, hyperosmolarity, inflammation, and epithelial dysfunction. This study evaluated the protective and therapeutic effects of NS2130E03, a standardized anthocyanin-rich purple carrot (Daucus carota subsp. sativus) extract, in a Benzalkonium Chloride (BAC)-induced murine model of DED. Male BALB/c mice (n=36) were randomized into six groups (n=6/group): normal control, BAC-induced pathological control, cyclosporine A (0.05%), and NS2130E03 at 50, 150, or 500mg/kg/day. NS2130E03 was administered orally for 14 days, while 0.2% BAC was applied topically for 7 consecutive days. Tear secretion, tear-film stability, and tear osmolarity were assessed using the Schirmer test, Tear Break-Up Time (TBUT), and TearLab osmometry, respectively.

Serum IL-1β, IL-6, TNF-α, and MUC5AC were quantified by ELISA, and ocular tissues were evaluated histopathologically. BAC significantly impaired tear secretion and tear-film stability and increased tear osmolarity. NS2130E03 significantly improved these parameters dose-dependently (P<0.001). At 500mg/kg, tear secretion and TBUT recovered to approximately 92% and 90% of normal-control values, respectively, while tear osmolarity decreased by 28.6% versus the pathological control. IL-1β, IL-6, and TNF-α decreased by 58.2%, 54.7%, and 57.6%, respectively, while MUC5AC increased by 81.2% (P<0.001). Histopathology demonstrated dose-dependent preservation of ocular tissue architecture. NS2130E03 attenuated BAC-induced DED by improving tear-film parameters, reducing inflammation, enhancing MUC5AC-associated responses, and preserving ocular tissue architecture. These findings support further investigation of anthocyanin-rich purple carrot extract as a potential adjunctive strategy for DED.

Keywords:Dry eye disease; NS2130E03; Purple carrot anthocyanins; Daucus carota; Benzalkonium chloride; Tear-film stability; Ocular inflammation; MUC5AC

Abbreviations: ANOVA: One-way Analysis of Variance; BAC: Benzalkonium Chloride; DED: Dry Eye Disease; H&E: Hematoxylin and eosin; IAEC: Institutional Animal Ethics Committee; IL-1β: Interleukin- 1β; INL: Inner Nuclear Layer; MAPK: Mitogen-Activated Protein Kinase; MUC5AC: Mucin-5 Subtype AC; NF-κB: Nuclear Factor-κB; ONL: Outer Nuclear Layer; PSL: Photoreceptor Segment Layer; RGC: Retinal Ganglion Cell; ROS: Reactive Oxygen Species; TBUT: Tear Break-Up Time; TNF-α: Tumor Necrosis Factor-α