The protein GSDMD is an important performer of pyroptosis and a universal substrate for the inflammatory caspase. However, the role and regulatory mechanism of GSDMD in Aspergillus fumigatus keratitis is remains unknown. Here we detected GSDMD protein in the cornea of normal and fungal-infected C57BL/6 mice. Human corneal epithelial cell (HCECs) were preincubated with a hydrochloride solution (IFNR inhibitor), ruxolitinib (JAK/STAT inhibitor), belnacasan (caspase-1 inhibitor) before infection with A. fumigatus conidia. Mice corneas were infected with Aspergillus fumigatus after pretreatment of GSDMD siRNA via subconjunctival injection. After, samples were harvested at specific time points and the expression of GSDMD and IL-1β was assessed by PCR, Western blot and immunofluorescence staining. Compared with the control group, we observed that the expression of GSDMD in fungal-infected mice cornea was significantly increased. After pretreatment with IFNR, JAK/STAT and caspase-1 inhibitors before fungal infection, the expression of GSDMD was significantly inhibited compared to the DMSO control in HCECs. Moreover, the GSDMD siRNA treatment have significantly weaken corneal inflammatory response, decreasing the proinflammatory factor IL-1β secretion and reducing neutrophils and macrophages recruitment in mice infected corneas. In summary, the data here provided evidences that GSDMD, an executor of pyroptosis, is involved in the early immune response of A. fumigatus keratitis. Additionally, the inhibition of GSDMD expression can affect the secretion of IL-1β and the recruitment of neutrophil and macrophages by blocking IFNR, JAK/STAT and caspase-1 signaling pathway. The protein GSDMD may emerge as a potential therapeutic target for A. fumigatus keratitis.

GSDMD蛋白是发烧的重要执行者，是炎症性胱天蛋白酶的通用底物。然而，GSMDD在烟曲霉性角膜炎中的作用和调节机制仍然未知。在这里，我们在正常和真菌感染的C57BL / 6小鼠的角膜中检测到GSDMD蛋白。在感染烟曲霉分生孢子之前，将人角膜上皮细胞（HCEC）与盐酸盐溶液（IFNR抑制剂），鲁索替尼（JAK / STAT抑制剂），贝拉卡森（半胱天冬酶-1抑制剂）预孵育。小鼠角膜感染了烟曲霉通过结膜下注射对GSDMD siRNA进行预处理后。之后，在特定时间点收集样品，并通过PCR，蛋白质印迹和免疫荧光染色评估GSMDD和IL-1β的表达。与对照组相比，我们观察到真菌感染的小鼠角膜中GSDMD的表达显着增加。在真菌感染前用IFNR，JAK / STAT和caspase-1抑制剂预处理后，与HCEC中的DMSO对照相比，GSDMD的表达被显着抑制。此外，GSDMD siRNA处理可显着减弱角膜炎性小鼠的角膜炎症反应，减少促炎因子IL-1β的分泌，并减少嗜中性粒细胞和巨噬细胞的募集。总而言之，此处的数据提供了证据，表明烧伤的执行者GSDMD烟曲霉角膜炎。另外，对GSDMD表达的抑制可通过阻断IFNR，JAK / STAT和caspase-1信号通路来影响IL-1β的分泌以及嗜中性粒细胞和巨噬细胞的募集。GSDMD蛋白可能成为烟曲霉性角膜炎的潜在治疗靶标。