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目的 通过蒙花苷(LA)对实验性干眼模型的干预,探讨LA对干眼的治疗效果和作用机制.方法 通过建立干眼人角膜上皮细胞干眼高渗模型和动物干眼模型,采用LA进行干预,用CCK-8实验评估LA对人角膜上皮细胞的毒性反应;对实验小鼠采用酚红棉线进行泪液量测量,采用0.1％荧光素钠染色条进行泪膜破裂时间的测定,采用俄勒冈绿(OGD)荧光染色检测角膜上皮的渗透功能;通过碘酸-希夫(PAS)染色对动物模型的结膜杯状细胞数量进行评估;通过原位末端转移酶标记技术(TUNEL)实验对LA对两种模型的细胞凋亡进行评估;通过免疫荧光染色对细胞模型的Ki-67、动物模型的MMP-3和MMP-9进行评估;采用RT-qPCR和Western blot对动物模型的NLRP3、ASC、Caspase-1、IL-1β、IL-18和TNF-α进行评估.结果 LA对人角膜上皮细胞无毒性反应,并在细胞高渗干眼模型下,LA能提高细胞的增殖反应,降低细胞的凋亡;LA能增加泪液分泌量和泪膜破裂时间、减少角膜OGD染色、增加结膜杯状细胞数量;LA能降低角结膜的MMP-3、MMP-9和TNF-α的表达,减少角膜上皮凋亡;LA能抑制动物干眼模型结膜的NLRP3、ASC、Caspase-1、IL-1β和IL-18的表达.结论 LA能安全、有效地通过改善干眼眼表屏障功能损伤、抑制眼表NLRP3炎症小体治疗干眼.“,”Objective To investigate the effect and mechanism of linarin (LA) in an experimental dry eye model. Methods LA or vehicle was applied in two dry eye models: an in vitro hyperosmotic stress model and an in vivo desiccating stress (DS) murine model. The viability of human corneal epithelial cells (HCECs) was measured using a cell counting kit (CCK-8). Tear secretion was assessed using the phenol red cotton test. The tear break-up time (TBUT) was recorded using 0.1％ liquid fluorescein sodium. Corneal epithelial permeability was evaluated through Oregon green dextran (OGD) staining. Conjunctival goblet cells were counted using periodic acid-Schiff (PAS) staining. Terminal deoxynucleotidyl transfer dUTP nick-end labeling (TUNEL) staining was used to quantify apoptotic cells in both models. The expression of Ki-67 was measured in HCECs in the cell model while that of matrix metalloproteinase (MMP)-3 and -9 was measured in the murine model through immunofluorescence staining. Real-time quantitative PCR (RT-qPCR) was performed to assess the expression of MMP-3 and MMP-9 in the corneal epithelium and NLRP3, ASC, Caspase-1, interleukin (IL)-1β, IL-18, and tumor necrosis factor (TNF)-α in the conjunctiva. The protein expression levels of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the conjunctiva were assessed via Western blot.Results In the in vitro model, treatment of HCECs with LA showed no toxicity, increased proliferation, and reduced apoptosis. In the murine model, compared to the control, LA significantly increased tear production and TBUT, improved OGD staining, and increased the number of goblet cells. Topical treatment of LA to mice provided decreased expression of MMP-3, MMP-9, TNF-α, and apoptotic corneal epithelium. Topical administration of LA also suppressed the NLRP3 inflammasome in the dry eye disease (DED) murine model by decreasing the expression of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the conjunctiva.Conclusion Our findings support the safety and efficacy of LA in the treatment of DED. LA alleviated corneal epithelial damage and suppressed NLRP3 inflammasome-mediated immunity in the conjunctiva in a murine model of DED.