活性氧在人晶状体上皮细胞上皮-间质转化及凋亡中的作用
Role of reactive oxygen species in epithelial-mesenchymal transition and apoptosis of human lens epithelial cells.
作者信息
Jing Rui-Hua, Hu Cong-Hui, Qi Tian-Tian, Ma Bo
机构信息
Department of Ophthalmology, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi Province, China.
Department of Ophthalmology, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
出版信息
Int J Ophthalmol. 2023 Dec 18;16(12):1935-1941. doi: 10.18240/ijo.2023.12.04. eCollection 2023.
AIM
To investigate the role of reactive oxygen species (ROS) in epithelial-mesenchymal transition (EMT) and apoptosis of human lens epithelial cells (HLECs).
METHODS
Flow cytometry was used to assess ROS production after transforming growth factor β2 (TGF-β2) induction. Apoptosis of HLECs after HO and TGF-β2 interference with or without ROS scavenger N-acetylcysteine (NAC) were assessed by flow cytometry. The corresponding protein expression levels of the EMT marker α-smooth muscle actin (α-SMA), the extracellular matrix (ECM), marker fibronectin (Fn), and apoptosis-associated proteins were detected by using Western blotting in the presence of an ROS scavenger (NAC). Wound-healing and Transwell assays were used to assess the migration capability of HLECs.
RESULTS
TGF-β2 stimulates ROS production within 8h in HLECs. Additionally, TGF-β2 induced HLECs cell apoptosis, EMT/ECM synthesis protein markers expression, and pro-apoptotic proteins production; nonetheless, NAC treatment prevented these responses. Similarly, TGF-β2 promoted HLECs cell migration, whereas NAC inhibited cell migration. We further determined that although ROS initiated apoptosis, it only induced the accumulation of the EMT marker α-SMA protein, but not COL-1 or Fn.
CONCLUSION
ROS contribute to TGF-β2-induced EMT/ECM synthesis and cell apoptosis of HLECs; however, ROS alone are not sufficient for EMT/ECM synthesis.
目的
探讨活性氧（ROS）在人晶状体上皮细胞（HLECs）上皮-间质转化（EMT）和凋亡中的作用。
方法
采用流式细胞术评估转化生长因子β2（TGF-β2）诱导后ROS的产生。通过流式细胞术评估在有或没有ROS清除剂N-乙酰半胱氨酸（NAC）的情况下，HO和TGF-β2干扰后HLECs的凋亡情况。在存在ROS清除剂（NAC）的情况下，使用蛋白质免疫印迹法检测EMT标志物α-平滑肌肌动蛋白（α-SMA）、细胞外基质（ECM）标志物纤连蛋白（Fn）以及凋亡相关蛋白的相应蛋白表达水平。采用划痕实验和Transwell实验评估HLECs的迁移能力。
结果
TGF-β2在8小时内刺激HLECs产生ROS。此外，TGF-β2诱导HLECs细胞凋亡、EMT/ECM合成蛋白标志物表达以及促凋亡蛋白产生；然而，NAC处理可阻止这些反应。同样，TGF-β2促进HLECs细胞迁移，而NAC抑制细胞迁移。我们进一步确定，虽然ROS引发凋亡，但它仅诱导EMT标志物α-SMA蛋白的积累，而不诱导COL-1或Fn的积累。
结论
ROS促成TGF-β2诱导的HLECs的EMT/ECM合成和细胞凋亡；然而，单独的ROS不足以促成EMT/ECM合成。
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