is a key regulator of eye development and epithelial homeostasis in the cornea. When deficient, chronic corneal inflammation, neovascularization and limbal stem cell deficiency can occur. Here we investigated the potential of duloxetine, a generic serotonin reuptake inhibitor that can upregulate PAX6 , for its activity in the context of corneal inflammation. Duloxetine tolerance was tested in a human limbal stem cell line and isogenic CRISPR-knockout cells. C57BL/6-Wildtype mice were administered duloxetine eye drops at concentrations of 1 μM - 2 mM and tested for toxicity and corneal PAX6 expression. In LPS-induced corneal inflammation in mice, duloxetine's effect on PAX6 expression, corneal opacification and inflammatory responses were evaluated by corneal imaging, immunostaining, and whole-transcriptome microarray analysis. No toxicity was observed for duloxetine concentrations up to 10μΜ. , duloxetine drops were well-tolerated up to 50 μM. Duloxetine drops at 10μΜ significantly upregulated PAX6 protein levels in the cornea by 30 % within 2 days. In the LPS model, duloxetine resulted in a sustained 33 % PAX6 protein upregulation in the cornea at 7 days, and in reduced opacity within 2 days, accompanied by a significant dampening of IL-17A signaling, neutrophil degranulation, microglial activation, macrophage markers, and expression, despite non-significant changes in total inflammatory cell infiltration. Short-term administration of a repurposed generic drug, duloxetine, upregulates PAX6 protein levels in the cornea of mice and exerts an anti-inflammatory activity by dampening innate immune responses.

中文翻译：

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度洛西汀增强 PAX6 表达并抑制小鼠 LPS 诱导的角膜炎症中的先天免疫反应


是眼睛发育和角膜上皮稳态的关键调节因子。当缺乏时，可能会发生慢性角膜炎症、新生血管形成和角膜缘干细胞缺乏。在这里，我们研究了度洛西汀（一种通用血清素再摄取抑制剂，可以上调 PAX6）在角膜炎症中的潜力。在人类角膜缘干细胞系和等基因 CRISPR 敲除细胞中测试了度洛西汀耐受性。对 C57BL/6-Wildtype 小鼠施用浓度为 1 μM - 2 mM 的度洛西汀滴眼液，并测试毒性和角膜 PAX6 表达。在 LPS 诱导的小鼠角膜炎症中，通过角膜成像、免疫染色和全转录组微阵列分析评估度洛西汀对 PAX6 表达、角膜混浊和炎症反应的影响。对于高达10μM的度洛西汀浓度，没有观察到毒性。 ，度洛西汀滴剂高达 50 μM 的耐受性良好。度洛西汀滴加 10μM 后，角膜中的 PAX6 蛋白水平在 2 天内显着上调 30%。在 LPS 模型中，度洛西汀导致角膜中 PAX6 蛋白在 7 天时持续上调 33%，并在 2 天内降低混浊度，同时显着抑制 IL-17A 信号传导、中性粒细胞脱颗粒、小胶质细胞激活、巨噬细胞标记物、和表达，尽管总炎症细胞浸润没有显着变化。短期服用一种经过改造的仿制药度洛西汀可以上调小鼠角膜中的 PAX6 蛋白水平，并通过抑制先天免疫反应发挥抗炎活性。