Centre for Ocular Research & Education, School of Optometry & Vision Science, University of Waterloo, Waterloo, Canada.
Centre for Eye and Vision Research (CEVR), Hong Kong SAR, China.
Curr Eye Res. 2022 Nov;47(11):1488-1495. doi: 10.1080/02713683.2022.2125531. Epub 2022 Sep 23.
To determine the metabolic activity, and cytokine expression over time from immortalized human corneal epithelial cells (HCECs) exposed to hyperosmotic stress.
HCECs were cultured and expanded in DMEM/F-12 with 10% FBS. The cells were exposed to either normal media (295 mmol/kg) or hyperosmolar media (500 mmol/kg) for 0.25, 3, 6, and 12 hours. After each exposure duration, metabolic activity was quantified using alamarBlue, and a panel of pro-inflammatory cytokines (IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, TNF-α, IFN-γ, and IL-17A) was quantified using multiplexed electrochemiluminescence (Meso Scale Diagnostics, Rockville, MD).
Metabolic activity of the HCEC exposed to hyperosmolar conditions was significantly reduced at the 3-, 6-, and 12-hour mark compared to the control (all < 0.01). There was no significant difference in cytokine expression between the hyperosmolar media and control at the 0.25- and 3-hour mark for all cytokines (all ≥ 0.28). The difference in cytokine expression between the hyperosmolar media and the control was significant for IL-1β, IL-4, IL-6, IL-8, IL-12p70, IL-13, and TNF-α at the 6-hour mark (all ≤ 0.02). No significant change in cytokine expression between the hyperosmolar media and control was noted for IL-2, IL-10, IL-17A, and IFN-γ (all ≥ 0.74) at the 6-hour mark.
Hyperosmolar stress reduced cell metabolic activity and increased expression of IL-1β, IL-4, IL6, IL8, IL-12p70, IL-13, and TNF-α over a 6-hour period in an immortalized HCEC line.
确定暴露于高渗环境下的永生化人角膜上皮细胞（HCEC）的代谢活性和细胞因子表达随时间的变化。
将 HCEC 在含有 10% FBS 的 DMEM/F-12 中培养和扩增。将细胞分别暴露于正常培养基（295mmol/kg）或高渗培养基（500mmol/kg）0.25、3、6 和 12 小时。在每个暴露时间后，使用 alamarBlue 定量测定代谢活性，并使用多重电化学发光（Meso Scale Diagnostics，Rockville，MD）定量测定一组促炎细胞因子（IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12p70、IL-13、TNF-α、IFN-γ 和 IL-17A）。
与对照组相比，暴露于高渗条件下的 HCEC 的代谢活性在 3、6 和 12 小时标记时明显降低（均 < 0.01）。在 0.25 和 3 小时标记时，所有细胞因子的高渗培养基与对照组之间的细胞因子表达均无显著差异（均 ≥ 0.28）。在 6 小时标记时，高渗培养基与对照组之间的细胞因子表达差异在 IL-1β、IL-4、IL-6、IL-8、IL-12p70、IL-13 和 TNF-α 方面具有统计学意义（均 ≤ 0.02）。在 6 小时标记时，高渗培养基与对照组之间的细胞因子表达差异在 IL-2、IL-10、IL-17A 和 IFN-γ 方面无统计学意义（均 ≥ 0.74）。
在永生化 HCEC 系中，高渗应激在 6 小时内降低了细胞代谢活性，并增加了 IL-1β、IL-4、IL6、IL8、IL-12p70、IL-13 和 TNF-α 的表达。
