| 刘 萱,张 婕,刘小毛,陈宁宁,刘 甜,解辽琦,赵吉飞.PM2.5对人角膜上皮细胞屏障功能、氧化应激反应以及自噬的影响[J].,2024,(22):4207-4213 |
| PM2.5对人角膜上皮细胞屏障功能、氧化应激反应以及自噬的影响 |
| Effects of PM2.5 on Barrier Function, Oxidative Stress Response and Autophagy of Human Corneal Epithelial Cell |
| 投稿时间：2024-04-22 修订日期：2024-05-16 |
| DOI：10.13241/j.cnki.pmb.2024.22.002 |
| 中文关键词: PM2.5 人角膜上皮细胞 屏障功能 氧化应激反应 细胞自噬 |
| 英文关键词: Particulate matter 2.5 (PM2.5) Human corneal epithelial cells (HCEC) Barrier function Oxidative stress response Autophagy |
| 基金项目:陕西省重点研发计划-国际科技合作计划项目（2022kw-43）；咸阳市重点研发计划（L2022ZDYFSF038） |
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| 中文摘要: |
| 摘要 目的：探讨PM2.5对人角膜上皮细胞屏障功能、氧化应激反应以及自噬的影响及其分子机制。方法：以不同浓度PM2.5处理人角膜上皮细胞（HCEC）24 h，采用CCK8检测细胞增殖，荧光标记法检测细胞内活性氧（ROS）水平，ELISA试剂盒检测IL-6、IL-8和TNF-α的含量；采用跨膜电阻实验（TEER）检测细胞的TEER变化，Western blot检测Beclin1、LC3 Ⅱ/Ⅰ、Claudin1、Occludin和ZO-1蛋白表达水平。再将HCEC细胞分为对照组、PM2.5组、NAC组和PM2.5+NAC组，所有组细胞培养24 h，后续进行细胞增殖水平、ROS、炎症因子、跨膜电阻值和Western blot检测。结果：与0 μg/mL比较，40 μg/mL、80 μg/mL、160 μg/mL和200 μg/mL的PM2.5处理的HCEC细胞相对增殖水平、TEER值和Claudin1、ZO-1和Occludin水平均降低（P<0.05），上清液中IL-6、IL-8和TNF-α水平均升高（P<0.05），HCEC细胞内ROS水平、Beclin-1和LC3 Ⅱ/Ⅰ蛋白水平均升高（P<0.05）。与PM2.5组比较，PM2.5+NAC组HCEC细胞相对增殖水平、TEER值和Claudin1、ZO-1和Occludin水平均升高（P<0.05），上清液中IL-6、IL-8和TNF-α水平均降低（P<0.05），HCEC细胞内ROS水平、Beclin-1和LC3 Ⅱ/Ⅰ蛋白水平均降低（P<0.05）。与NAC组比较，PM2.5+NAC组HCEC细胞相对增殖水平、TEER值和Claudin1、ZO-1和Occludin水平均降低（P<0.05），上清液中IL-6、IL-8和TNF-α水平均升高（P<0.05），HCEC细胞内ROS相对水平、Beclin-1和LC3 Ⅱ/Ⅰ蛋白水平均升高（P<0.05）。结论：PM2.5通过诱导炎症和氧化应激反应的产生，破坏人角膜上皮细胞的屏障功能，并促进细胞自噬。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of PM2.5 on barrier function, oxidative stress response and autophagy of human corneal epithelial cell and its molecular mechanism. Methods: Human corneal epithelial cells (HCEC) were treated with different concentrations of PM2.5 for 24 h. The cell proliferation level was detected by CCK8. Intracellular reactive oxygen species (ROS) were detected by fluorescent labeling. The levels of IL-6, IL-8 and TNF-α were detected by ELISA kit. The changes of TEER in HCEC cells were detected by transmembrane resistance assay (TEER). The protein expression levels of Beclin1, LC3 Ⅱ/I, Claudin1, Occludin and ZO-1 were detected by Western blot. Then HCEC cells were divided into control group, PM2.5 group, NAC group and PM2.5+NAC group, and the cells of all groups were cultured for 24 h. Cell proliferation level, ROS, inflammatory factors, transmembrane resistance and Western blot were subsequently detected. Results: Compared with 0 μg/mL, the relative proliferation level, TEER value, and the levels of Claudin1, ZO-1 and Occludin protein of HCEC cells treated with 40 μg/mL, 80 μg/mL, 160 μg/mL and 200 μg/mL PM2.5 were decreased (P<0.05), and the levels of IL-6, IL-8 and TNF-α in supernatant were increased (P<0.05). The level of ROS and the protein levels of Beclin-1 and LC3 Ⅱ/Ⅰ in HCEC cells were increased (P<0.05). Compared with the PM2.5 group, the relative proliferation level, TEER value, and the levels of Claudin1, ZO-1 and Occludin protein of HCEC cells in PM2.5+NAC group was increased (P<0.05), the levels of IL-6, IL-8 and TNF-α in supernatant were decreased (P<0.05), and the level of ROS and the protein levels of Beclin-1 and LC3 Ⅱ/Ⅰ in HCEC cells were decreased (P<0.05). Compared with the NAC group, the relative proliferation level, TEER value, and the levels of Claudin1, ZO-1 and Occludin protein of HCEC cells in PM2.5+NAC group was decreased (P<0.05), the levels of IL-6, IL-8 and TNF-α in supernatant were increased (P<0.05), and the level of ROS and the protein levels of Beclin-1 and LC3 Ⅱ/Ⅰ in HCEC cells were increased (P<0.05). Conclusion: PM2.5 damages the barrier function of human corneal epithelial cells and promotes autophagy by inducing inflammation and oxidative stress responses. |
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