Wang Yuzhou, Xu Zhiqiang, Wei Linzhi, Lu Yang, Shi Yizhou, Wen Shiyu, Lv Xiujuan, Huang Kaiyan, Lu Fan, Qu Jia, Hu Liang
National Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
Invest Ophthalmol Vis Sci. 2025 Apr 1;66(4):28. doi: 10.1167/iovs.66.4.28.
This study aimed to investigate the protective effects of keratinocyte growth factor-2 (KGF-2) in dry eye disease (DED) and elucidate its mechanism of action through the regulation of the HMGB1/TLR4 pathway.
Two in vitro models were established by stimulating hyperosmolar human corneal epithelial cells (HCECs) and RAW 264.7 cells with lipopolysaccharide. A DED mice model was established using scopolamine and an intelligently controlled environmental system. After KGF-2 treatment, the symptoms of the DED mice were assessed. The changes in inflammatory factors were measured using Western blotting and quantitative reverse-transcription polymerase chain reaction (RT-qPCR). RNA sequencing (RNA-seq) was used to identify the key factors involved in KGF-2 treatment, followed by validation through in vivo and in vitro knockdown of the relevant factors.
KGF-2 treatment significantly relieved DED in the mice model through increased tear secretion, and improved fluorescein staining scores. In addition, the levels of inflammatory factors were effectively lowered in both in vitro and in vivo models. Bulk RNA-seq analysis suggested that KGF-2 exerts its effects by regulating the HMGB1/TLR4 pathway. Furthermore, KGF-2 treatment inhibited the upregulation and nuclear translocation of HMGB1 in the DED model, thereby suppressing the levels of inflammatory factors associated with the HMGB1/TLR4 pathway. Knockdown of HMGB1 in HCECs and glycyrrhizin treatment in DED mice exhibited therapeutic effects similar to those of KGF-2.
KGF-2 demonstrated protective effects in both in vivo and in vitro DED models by modulating the HMGB1/TLR4 pathway. These findings suggest its potential as a therapeutic agent for DED, warranting further clinical investigation in this regard.
本研究旨在探讨角质形成细胞生长因子-2（KGF-2）在干眼症（DED）中的保护作用，并通过调节高迁移率族蛋白B1（HMGB1）/Toll样受体4（TLR4）通路阐明其作用机制。
通过用脂多糖刺激高渗人角膜上皮细胞（HCECs）和RAW 264.7细胞建立两种体外模型。使用东莨菪碱和智能控制环境系统建立DED小鼠模型。给予KGF-2治疗后，评估DED小鼠的症状。采用蛋白质免疫印迹法和定量逆转录聚合酶链反应（RT-qPCR）检测炎症因子的变化。采用RNA测序（RNA-seq）确定KGF-2治疗涉及的关键因子，随后通过体内和体外敲低相关因子进行验证。
KGF-2治疗通过增加泪液分泌显著缓解了小鼠模型中的DED，并改善了荧光素染色评分。此外，在体外和体内模型中炎症因子水平均有效降低。全转录组RNA测序分析表明，KGF-2通过调节HMGB1/TLR4通路发挥作用。此外，KGF-2治疗抑制了DED模型中HMGB1的上调和核转位，从而抑制了与HMGB1/TLR4通路相关的炎症因子水平。在HCECs中敲低HMGB1以及在DED小鼠中给予甘草酸治疗均表现出与KGF-2相似的治疗效果。
KGF-2通过调节HMGB1/TLR4通路在体内和体外DED模型中均显示出保护作用。这些发现表明其作为DED治疗药物的潜力，值得在这方面进行进一步的临床研究。
