To investigate the role of miR-129-5p in inflammation and autophagy in fungal keratitis, we established a keratitis mouse model infected with Fusarium solani (F. solani) and conducted experiments on corneal stromal cells infected with F. solani. The expression of miR-129-5p was detected via quantitative real-time polymerase chain reaction (PCR). The miR-129-5p antagomir was used to transfect cells and mice to study the regulatory role of miR-129-5p in autophagy and inflammation after fungal infection. The expression of Beclin1 and LC3B and colocalization of LC3B with lysosomes were detected via Western blotting and immunofluorescence. CCK-8 was used to determine the viability of corneal stromal cells. The expression of IL-1β were detected by ELISA. Bioinformatics software was used to predict the potential targets of miR-129-5p, which were verified by a luciferase reporter gene assay. RT-PCR showed that miR-129-5p expression in mouse corneas was significantly increased after infection with F. solani. Subconjunctival injection of the miR-129-5p antagomir significantly enhanced the proteins Beclin-1 and LC3B. At the same time, inhibiting miR-129-5p expression could reduce the inflammatory response in FK and significantly increase the viability of corneal stromal cells infected with F. solan. Moreover, the dual luciferase reporter assay indicated that Atg14 was a direct target of miR-129-5p. Our study shows that miR-129-5p is a novel small molecule that regulates autophagy by targeting Atg14, indicating that it may be a proinflammatory and therapeutic target for fungal keratitis.

为了研究miR-129-5p在真菌性角膜炎炎症和自噬中的作用，我们建立了茄病镰刀菌（F. solani）感染的角膜炎小鼠模型，并对茄病镰刀菌感染的角膜基质细胞进行了实验。. 通过定量实时聚合酶链反应 (PCR) 检测 miR-129-5p 的表达。用miR-129-5p antagomir转染细胞和小鼠，研究miR-129-5p在真菌感染后自噬和炎症中的调节作用。通过蛋白质印迹和免疫荧光检测 Beclin1 和 LC3B 的表达以及 LC3B 与溶酶体的共定位。CCK-8 用于确定角膜基质细胞的活力。ELISA检测IL-1β的表达。使用生物信息学软件预测 miR-129-5p 的潜在靶点，并通过荧光素酶报告基因检测进行验证。RT-PCR结果显示，感染F. solani后小鼠角膜中miR-129-5p表达显着增加. 结膜下注射 miR-129-5p antagomir 显着增强了 Beclin-1 和 LC3B 蛋白。同时，抑制 miR-129-5p 的表达可以降低 FK 的炎症反应，并显着增加感染F. solan的角膜基质细胞的活力。此外，双荧光素酶报告基因检测表明 Atg14 是 miR-129-5p 的直接靶标。我们的研究表明，miR-129-5p 是一种通过靶向 Atg14 调节自噬的新型小分子，表明它可能是真菌性角膜炎的促炎和治疗靶点。