Chen Lirong, Gu Cao, Yang Yaling, He Taiwen, Zhang Qi
Department of Ophthalmology, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
Fundus Department, Shanghai New Vision Zhongxing Ophthalmology Hospital, Shanghai, China.
Exp Eye Res. 2025 Sep;258:110490. doi: 10.1016/j.exer.2025.110490. Epub 2025 Jun 18.
Dry eye disease negatively impacts the quality of life of many patients worldwide, and currently, there is no cure. Stem cell therapy may offer a potential new treatment option. Mesenchymal stem cells (MSCs)-derived exosomes have similar effects as MSCs but with fewer side effects. This study investigated the effects and mechanisms of human umbilical cord mesenchymal stem cells (HUCMSCs)-derived exosomes in cell and mouse models of dry eye disease. Exosomes were isolated from HUCMSCs (HUCMSCs-EXO) and characterized by measuring surface markers. Human corneal epithelial cells (HCECs) were cultured in hypertonic (500 mOsm) or isotonic medium (310 mOsm) and treated with HUCMSC-EXO or PBS. A mouse model of dry eye disease was generated by treating mice with benzalkonium chloride and confirmed by measuring tear secretion and performing H&E staining. Cell viability, apoptosis, RNA, and protein expression levels were assessed using CCK-8 assay, TUNEL staining, qPCR, and Western blotting. The regulation of SQSTM1 expression by miR-146a was evaluated using a dual luciferase reporting assay. Exposure to hyperosmotic pressure decreased cell viability, increased apoptosis and inflammation, decreased miR-146a expression, and increased SQSTM1 expression in HCECs. Treatment with HUCMSCs-EXO alleviated the effects of hyperosmotic pressure on inflammation and apoptosis. Overexpression of miR-146a increased cell viability and inhibited apoptosis and inflammation, suggesting that miR-146a mediates the beneficial effects of HUCMSCs-EXO. These findings were validated in the mouse model of dry eye disease. Further experiments revealed that miR-146a targets and promotes SQSTM1 expression. Overexpression of SQSTM1 increased cell viability and inhibited apoptosis and inflammation in HCECs. In conclusion, this study demonstrated that HUCMSCs-derived exosomal miR-146a targets SQSTM1 and promotes its expression, resulting in the alleviation of inflammation and apoptosis in cell and mouse models of dry eye disease.
干眼症对全球许多患者的生活质量产生负面影响，目前尚无治愈方法。干细胞疗法可能提供一种潜在的新治疗选择。间充质干细胞（MSCs）衍生的外泌体具有与MSCs相似的作用，但副作用较少。本研究调查了人脐带间充质干细胞（HUCMSCs）衍生的外泌体在干眼症细胞和小鼠模型中的作用及机制。从HUCMSCs中分离出外泌体（HUCMSCs-EXO），并通过测量表面标志物进行表征。将人角膜上皮细胞（HCECs）培养在高渗（500 mOsm）或等渗培养基（310 mOsm）中，并用HUCMSC-EXO或PBS处理。通过用苯扎氯铵处理小鼠建立干眼症小鼠模型，并通过测量泪液分泌和进行苏木精-伊红染色进行确认。使用CCK-8法、TUNEL染色、qPCR和蛋白质印迹法评估细胞活力、凋亡、RNA和蛋白质表达水平。使用双荧光素酶报告试验评估miR-146a对SQSTM1表达的调控。暴露于高渗压力会降低HCECs的细胞活力，增加凋亡和炎症，降低miR-146a表达，并增加SQSTM1表达。用HUCMSCs-EXO处理可减轻高渗压力对炎症和凋亡的影响。miR-146a的过表达增加了细胞活力并抑制了凋亡和炎症，表明miR-146a介导了HUCMSCs-EXO的有益作用。这些发现在干眼症小鼠模型中得到了验证。进一步的实验表明，miR-146a靶向并促进SQSTM1表达。SQSTM1的过表达增加了HCECs的细胞活力并抑制了凋亡和炎症。总之，本研究表明，HUCMSCs衍生的外泌体miR-146a靶向SQSTM1并促进其表达，从而减轻干眼症细胞和小鼠模型中的炎症和凋亡。
Comb Chem High Throughput Screen. 2024-5-15
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