透明质酸通过 CD44 和 TGF-β2 介导人晶状体上皮细胞的上皮-间充质转化。
Hyaluronic acid-mediated epithelial-mesenchymal transition of human lens epithelial cells via CD44 and TGF-β2.
机构信息
Wuhan Puren Hospital, Qingshan District Jianshe Fourth Road Benxi Street Te #1, Wuhan, Hubei Province, China.
Wuhan University Renmin Hospital, Wuchang District Ziyang Road 22, Wuhan, Hubei Province, China.
出版信息
Arq Bras Oftalmol. 2024 Aug 2;88(1):e20230163. doi: 10.5935/0004-2749.2023-0163. eCollection 2024.
PURPOSE
The epithelial-mesenchymal transition of human lens epithelial cells plays a role in posterior capsule opacification, a fibrotic process that leads to a common type of cataract. Hyaluronic acid has been implicated in this fibrosis. Studies have investigated the role of transforming growth factor (TGF)-β2 in epithelial-mesenchymal transition. However, the role of TGF-β2 in hyaluronic acid-mediated fibrosis of lens epithelial cell remains unknown. We here examined the role of TGF-β2 in the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells.
METHODS
Cultured human lens epithelial cells (HLEB3) were infected with CD44-siRNA by using the Lipofectamine 3000 transfection reagent. The CCK-8 kit was used to measure cell viability, and the scratch assay was used to determine cell migration. Cell oxidative stress was analyzed in a dichloro-dihydro-fluorescein diacetate assay and by using a flow cytometer. The TGF-β2 level in HLEB3 cells was examined through immunohistochemical staining. The TGF-β2 protein level was determined through western blotting. mRNA expression levels were determined through quantitative real-time polymerase chain reaction.
RESULTS
Treatment with hyaluronic acid (1.0 μM, 24 h) increased the epithelial-mesenchymal transition of HLEB3 cells. The increase in TGF-β2 levels corresponded to an increase in CD44 levels in the culture medium. However, blocking the CD44 function significantly reduced the TGF-β2-mediated epithelial-mesenchymal transition response of HLEB3 cells.
CONCLUSIONS
Our study showed that both CD44 and TGF-β2 are critical contributors to the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells, and that TGF-β2 in epithelial-mesenchymal transition is regulated by CD44. These results suggest that CD44 could be used as a target for preventing hyaluronic acid-induced posterior capsule opacification. Our findings suggest that CD44/TGF-β2 is crucial for the hyaluronic acid-induced epithelial-mesenchymal transition of lens epithelial cells.
目的
人晶状体上皮细胞的上皮-间充质转化在后囊混浊中起作用，后囊混浊是一种导致常见白内障的纤维化过程。透明质酸与此纤维化有关。研究已经探讨了转化生长因子（TGF）-β2 在晶状体上皮细胞上皮-间充质转化中的作用。然而，TGF-β2 在透明质酸介导的晶状体上皮细胞纤维化中的作用尚不清楚。我们在此研究了 TGF-β2 在透明质酸介导的晶状体上皮细胞上皮-间充质转化中的作用。
方法
用 Lipofectamine 3000 转染试剂将 CD44-siRNA 感染到培养的人晶状体上皮细胞（HLEB3）中。用 CCK-8 试剂盒测量细胞活力，用划痕实验测定细胞迁移。通过二氯二氢荧光素二乙酸酯测定和流式细胞仪分析细胞氧化应激。通过免疫组织化学染色检查 HLEB3 细胞中的 TGF-β2 水平。通过 Western 印迹测定 TGF-β2 蛋白水平。通过定量实时聚合酶链反应测定 mRNA 表达水平。
结果
用透明质酸（1.0 μM，24 小时）处理增加了 HLEB3 细胞的上皮-间充质转化。培养物中 TGF-β2 水平的增加与 CD44 水平的增加相对应。然而，阻断 CD44 功能显著降低了 HLEB3 细胞中 TGF-β2 介导的上皮-间充质转化反应。
结论
我们的研究表明，CD44 和 TGF-β2 都是晶状体上皮细胞透明质酸介导的上皮-间充质转化的关键贡献者，上皮-间充质转化中的 TGF-β2 受 CD44 调节。这些结果表明 CD44 可作为预防透明质酸诱导的后囊混浊的靶点。我们的研究结果表明，CD44/TGF-β2 对透明质酸诱导的晶状体上皮细胞上皮-间充质转化至关重要。
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