Dopamine Receptor 1 Treatment Promotes Epithelial Repair of Corneal Injury by Inhibiting NOD-Like Receptor Protein 3-Associated Inflammation

PURPOSE. To elucidate the influence of dopamine receptor 1 (DRD1) on the proliferation of mouse corneal epithelial cells (MCECs) under inflammatory conditions. METHODS. In vitro, immortalized MCECs (iMCECs) were treated with IL -1/3, with and without pcDNA3.1_DRD1. Primary MCECs (pMCECs) were exposed to IL -1/3, with and without DRD1 agonist (A68930). Cell proliferation was quantified using the Cell Counting Kit -8 (CCK-8) assay and immunofluorescence staining for Ki-67 and p63. Expression levels of NOD -like receptor protein 3 (NLRP3), IL -1/3, and IL -6 were assessed. To establish a corneal injury model in mice, a 2 -mm superficial keratectomy was performed. Either 0.1% A68930 or PBS was topically administered three times daily to the injured eyes for up to 5 days post -injury. Immunofluorescence analysis was employed to evaluate the expression of Ki-67, p63, and CD45 in mouse corneas. Western blotting and real-time quantitative PCR were utilized for quantitative analysis of DRD1, NLRP3, IL -1/3, and IL -6 in mouse corneas. Corneal epithelial regeneration was monitored through fluorescein sodium staining for a duration of up to 5 days following the injury. RESULTS. Overexpression of DRD1 and A68930 promoted MCEC proliferation and suppressed the expression of NLRP3, IL -1/3, and IL -6 in vitro. Topical application of the 0.1% A68930 following mechanical corneal injury in mice led to increased Ki-67 and p63 expression compared to PBS treatment. Furthermore, topical administration of the 0.1% A68930 reduced the expression of CD45, NLRP3, IL -1/3, and IL -6. Analysis with fluorescein sodium indicated accelerated corneal epithelial regeneration in the 0.1% A68930 treatment group. CONCLUSIONS. DRD1 treatment counteracts NLRP3-associated inflammation and facilitates epithelial repair of corneal injury.