Given the escalating global prevalence of dry eye disease (DED) and the inadequacy of current palliative management, this study was to evaluate thetherapeutic potential of recombinant lysozyme (rlysozyme) and investigate underlying mechanisms. Human corneal epithelial cells (HCEC) were subjected to hyperosmotic stress and treated with graded rlysozyme. Cell viability, wound healing, barrier integrity, oxidative stress, cytokines, and mitochondrial function were evaluated by MTS and scratch assays, immunofluorescent staining, ELISA, qRT-PCR, and flow cytometry. Results: No cytotoxicity was observed at any of the tested concentrations (2.76 × 10⁻⁹–2.76 × 10⁻⁴ M) in HCEC. In hyperosmolarity-induced DED cells, rlysozyme at the highest concentrations (2.76 × 10⁻⁴ M) significantly increased cell viability by 12.30% (p < 0.001) and promoted corneal repair by 45.04% (p < 0.05), which were comparable to those of 0.4% hyaluronic acid (the positive control). It also improved via restoring ZO-1 organization. Mechanistically, qRT-PCR results revealed that rlysozyme reduced mRNA expression of IL-1β (by 28.7%), TNF (by 39.2%) and key NLRP3 inflammasome components, including NLRP3 (by 62.0%) and PYD and CARD domain-containing protein gene(PYCARD, 21.4%), while caspase-1 (CASP1) and IL-6 expression remained unchanged. Moreover, oxidative stress was attenuated, as evidenced by significant decreases in reactive oxygen species (ROS, by 59.1%), 8-hydroxy-2-deoxyguanosine levels (8-OHdG, by 23.6%) and total matrix metalloproteinase-9 (MMP-9, by 64.3%) protein levels (p < 0.001), while mitochondrial membrane potential was assessed qualitatively. In conclusion, this study demonstrates thatrlysozyme effectively enhanced corneal epithelial cell viability, promoted wound healing, and restored ZO-1 structural organization without inducing cytotoxicity. These protective effects were associated with downregulation of NLRP3 inflammasome-related genes and attenuation of oxidative stress, supporting further investigation of its therapeutic potential in DED.

Dry eye disease (DED) is a prevalent ocular surface disorder with diverse clinical presentations, yet the comparative efficacy of various Traditional Chinese Medicine (TCM) formulations remains unclear. This systematic review and network meta-analysis evaluated the comparative effects and baseline response relationships of TCM classes for DED across 33 randomized controlled trials retrieved from PubMed, Embase, Cochrane Library, CINAHL, and CNKI. Primary outcomes included Schirmer I test (SIT), tear film break-up time (TBUT), and corneal fluorescein staining (FL). Random-effects models estimated weighted mean differences (WMD) with 95% credible intervals (CrIs); for FL, which was measured on heterogeneous grading scales and standardized mean differences (SMD) were used. TCM classes exhibited outcome-specific profiles: Yin–Kidney (YK) and Qi–Yin Support (QYS) produced the greatest improvements in SIT versus standard care (YK: WMD 2.56, 95% CrI 0.25 to 4.86; QYS: WMD 5.32, 95% CrI 0.63 to 10.06) and significantly improved TBUT (YK: WMD 2.04, 95% CrI 1.12 to 2.89; YM: WMD 2.49, 95% CrI 0.64 to 4.37). YK demonstrated the greatest FL reduction (SMD −0.37, 95% CrI −0.71 to 0.00), followed by Liver–Spleen Regulating (SMD −0.30, 95% CrI −0.99 to 0.39). Meta-regression revealed that patients with more severe baseline deficits in tear secretion (lower SIT) and more extensive ocular-surface damage (higher FL) achieved greater clinical improvements, whereas baseline of TBUT did not significantly moderate treatment response. Overall, TCM serves as an effective adjunctive therapy for DED, with selection guided by clinical pattern and target outcomes.