IGFBP-3 Mediates Metabolic Homeostasis During Hyperosmolar Stress in the Corneal Epithelium.
IGFBP-3 Mediates Metabolic Homeostasis During Hyperosmolar Stress in the Corneal Epithelium.
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DOI：
10.1167/iovs.62.7.11
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发表时间：
2021-06-01
影响因子：
4.4
通讯作者：
Robertson DM
中科院分区：
文献类型：
--
作者：
Bogdan ED;Stuard WL;Titone R;Robertson DM
胰岛素样生长因子结合蛋白-3（IGFBP-3）是一种多功能分泌蛋白，在细胞生长和存活中具有众所周知的作用。我们实验室的数据表明IGFBP-3可能在角膜上皮中发挥应激反应蛋白的作用。本研究的目的是确定IGFBP-3在介导角膜上皮细胞对高渗透压的应激反应中的作用，高渗透压是干眼病发展中的一种众所周知的病理生理学事件。端粒酶永生化的人角膜上皮（hTCEpi）细胞在这项研究中使用。细胞在含有（生长）或不含（基础）补充物的无血清培养基中培养。通过将盐浓度增加至450和500 mOsM来实现高渗透压。代谢和线粒体的变化进行了评估海马代谢通量分析和线粒体钙，极化和mtDNA的测定。使用ELISA定量IGFBP-3和炎症介质的水平。采用乳酸脱氢酶试验评价细胞毒性。在选择的实验中，用500 ng/mL重组人（rh）IGFBP-3共处理细胞。高渗性应激改变了代谢活性，使细胞向呼吸表型转变。高渗性应激进一步改变线粒体钙水平，使线粒体去极化，降低ATP、mtDNA水平和IGFBP-3表达。相反，高渗性应激增加促炎细胞因子IL-6和IL-8的产生。补充rhIGFBP-3消除了代谢和线粒体的变化，对IL-8只有轻微的影响。这些发现表明IGFBP-3是参与角膜上皮高渗应激反应的关键蛋白。这些数据进一步支持IGFBP-3在控制细胞代谢中的新作用。
The insulin-like growth factor binding protein-3 (IGFBP-3) is a multifunctional secretory protein with well-known roles in cell growth and survival. Data in our laboratory suggest that IGFBP-3 may be functioning as a stress response protein in the corneal epithelium. The purpose of this study is to determine the role of IGFBP-3 in mediating the corneal epithelial cell stress response to hyperosmolarity, a well-known pathophysiological event in the development of dry eye disease. Telomerase-immortalized human corneal epithelial (hTCEpi) cells were used in this study. Cells were cultured in serum-free media with (growth) or without (basal) supplements. Hyperosmolarity was achieved by increasing salt concentrations to 450 and 500 mOsM. Metabolic and mitochondrial changes were assessed using Seahorse metabolic flux analysis and assays for mitochondrial calcium, polarization and mtDNA. Levels of IGFBP-3 and inflammatory mediators were quantified using ELISA. Cytotoxicity was evaluated using a lactate dehydrogenase assay. In select experiments, cells were cotreated with 500 ng/mL recombinant human (rh)IGFBP-3. Hyperosmolar stress altered metabolic activity, shifting cells towards a respiratory phenotype. Hyperosmolar stress further altered mitochondrial calcium levels, depolarized mitochondria, decreased levels of ATP, mtDNA, and expression of IGFBP-3. In contrast, hyperosmolar stress increased production of the proinflammatory cytokines IL-6 and IL-8. Supplementation with rhIGFBP-3 abrogated metabolic and mitochondrial changes with only marginal effects on IL-8. These findings indicate that IGFBP-3 is a critical protein involved in hyperosmolar stress responses in the corneal epithelium. These data further support a new role for IGFBP-3 in the control of cellular metabolism.
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