IL-1b 介导的角膜上皮细胞免疫代谢适应性

Jose Marcos Sanches, Rajalakshmy Ayilam Ramachandran, Natalia Mussi, Hamid Baniasadi, Danielle M Robertson
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摘要

目的:作为外部粘膜表面,角膜上皮要承受一系列已知会引发炎症的压力。IL-1b是炎症的主要调节因子,由眼表上皮细胞和浸润的免疫细胞分泌到眼前泪膜中。虽然 IL-1b 水平的升高与角膜疾病有关,但 IL-1b 对角膜上皮细胞(CECs)线粒体功能的影响尚不清楚:为了研究 IL-1b 对线粒体功能的影响,将端粒酶永生化的人 CECs 在 50 纳克/毫升或 100 纳克/毫升 IL-1b 中进行短期(24 小时)或长期(72 小时)培养。对细胞的 ROS、炎症细胞因子的产生、线粒体极化和超微结构、有丝分裂以及代谢物组成的变化进行评估。使用光镜和荧光显微镜检查脂滴:结果:短期暴露于IL-1b会引发IL-8和ROS水平的增加,这与线粒体膜电位的降低相对应。长期暴露也显示 IL-8 和 IL-6 水平升高,ROS 水平进一步升高。然而,在长期接触后,线粒体膜电位出现了矛盾性的增加,这与剩余呼吸能力的增加和线粒体过度融合有关。代谢组学证实了磷酸戊糖途径和 TCA 循环的上调。富马酸也有所增加,表明通过复合体 II 的通量增加。脂质代谢的变化包括心磷脂和新生三酰甘油生物合成的上调,以及脂滴数量的增加。结论长期暴露于 IL-1b 可诱导 CECs 的新陈代谢重新布线,从而提高剩余呼吸能力。这些研究结果表明,角膜上皮能够适应一定程度的慢性炎症,对我们了解眼表上皮的免疫调节和细胞应激反应具有重要意义。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
IL-1b-mediated Immunometabolic Adaptation in Corneal Epithelial Cells
Purpose: As an external mucosal surface, the corneal epithelium is subject to a barrage of stressors that are known to trigger inflammation. IL-1b, a master regulator of inflammation, is secreted into the preocular tear film by ocular surface epithelial cells and infiltrating immune cells. While increased levels of IL-1b have been associated with corneal disease, the effects of IL-1b on mitochondrial function in corneal epithelial cells (CECs) is unknown. Methods: To investigate the effects of IL-1b on mitochondrial function, telomerase immortalized human CECs were cultured in either 50 ng/mL or 100 ng/mL IL-1b for short term (24 hours) or prolonged (72 hours) time periods. Cells were assessed for ROS, inflammatory cytokine production, mitochondrial polarization and ultrastructure, mitophagy, and changes in the metabolite composition. Lipid drops were examined using light and fluorescent microscopy. Results: Short term exposure to IL-1b triggered an increase in IL-8 and ROS levels that corresponded to a reduction in mitochondrial membrane potential. Long term exposure also showed increased levels of IL-8 and IL-6 and further increased ROS. After long term exposure however, there was a paradoxical increase in mitochondrial membrane potential that was associated an increase in spare respiratory capacity and mitochondrial hyperfusion. Metabolomics confirmed an upregulation of the pentose phosphate pathway and the TCA cycle. Fumarate was also increased, suggesting an increase in flux through complex II. Changes in lipid metabolism included an upregulation in cardiolipin and de novo triacylglyceride biosynthesis, along with increasing numbers of lipid droplets. Conclusion: Prolonged exposure to IL-1b induces metabolic rewiring in CECs that results in an increase in spare respiratory capacity. These findings suggest that the corneal epithelium is able to adapt to certain levels of chronic inflammation and may have important implications in our understanding of immune tone and cellular stress responses in ocular surface epithelia.
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