Ethanol-exposed lung fibroblasts cause airway epithelial barrier dysfunction.
Ethanol-exposed lung fibroblasts cause airway epithelial barrier dysfunction.
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DOI：
10.1111/acer.15174
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发表时间：
2023-10
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影响因子：
--
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慢性酒精摄入易导致败血症时肺损伤和修复功能丧失。我们以前的研究概述了转化生长因子-β1(转化生长因子-β1)和粒-巨噬细胞集落刺激因子(GM-β)在上皮屏障动态平衡中的作用，以及酒精如何扰乱它们的表达和信号。在这里，我们假设酒精暴露的肺成纤维细胞(LF)是转化生长因子β1和GM-CSF失调源，从而改变呼吸道上皮屏障功能。人或大鼠LF经±乙醇培养2周后与接种于Transwell通透性支架上的人或大鼠呼吸道上皮细胞(AEC)共培养。在选定的组中，应用转化生长因子β1受体1型(转化生长因子βR1)抑制剂(SB431542)或转化生长因子β1中和抗体。在共培养前和共培养第5天测量跨皮细胞电阻(TER)。然后分析血管内皮细胞紧密连接和间充质蛋白的表达，并用免疫荧光显微镜分析跨膜ZO-1的表达和定位。用双抗体夹心ELISA法测定条件培养液中转化生长因子β-1和GM-CSF值。与乙醇联合培养的血管内皮细胞，TER显著降低，ZO-1表达相应减少，而1A1型胶原和α-平滑肌肌动蛋白表达增加。同时，在ELF+AEC共培养的条件培养液中，活化的转化生长因子β1水平升高，GM-CSF1水平下降。值得注意的是，ELF对血管内皮细胞的所有作用均可通过阻断转化生长因子β1的活性而被阻止。乙醇预先暴露于LF可通过激活转化生长因子β1信号通路和抑制GM-CSF1，以旁分泌方式诱导幼稚血管内皮细胞屏障功能障碍。这些实验发现提供了一种潜在的机制，通过这种机制，慢性酒精摄入损害了呼吸道上皮细胞的完整性，使个人容易受到肺损伤。
Chronic alcohol ingestion predisposes to lung injury and disrepair during sepsis. Our previous studies outlined roles for transforming growth factor-beta 1 (TGFβ1) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in epithelial barrier homeostasis and how alcohol perturbs their expression and signaling. Here we hypothesize that ethanol-exposed lung fibroblasts (LF) are a source of dysregulated TGFβ1 and GM-CSF and thereby alter airway epithelial barrier function. Human or rat LF were cultured ± ethanol for 2 weeks and then co-cultured with human or rat airway epithelial cells (AEC) seeded on Transwell permeable supports. In selected groups, a TGFβ1 receptor type 1 (TGFβR1) inhibitor (SB431542) or a TGFβ1 neutralizing antibody was applied. Transepithelial electrical resistance (TER) was measured prior to co-culture and on day 5 of co-culture. AEC were then analyzed for the expression of selected tight junction and mesenchymal proteins, and transwell membranes were analyzed by immunofluorescence microscopy for ZO-1 expression and localization. TGFβ1 and GM-CSF levels in conditioned media from the co-cultures were quantified by ELISA. AEC co-cultured with ethanol-exposed LF (ELF) showed a significant reduction in TER and corresponding decreases in ZO-1 expression, whereas collagen type 1A1 and α-smooth muscle actin protein expression were increased. In parallel, in conditioned media from the ELF + AEC co-cultures, activated TGFβ1 levels increased and GM-CSF levels decreased. Notably, all the effects of ELF on the AEC were prevented by blocking TGFβ1 activity. Prior ethanol exposure to LF induces barrier dysfunction in naive AEC in a paracrine fashion through activation of TGFβ1 signaling and suppression of GM-CSF. These experimental findings provide a potential mechanism by which chronic alcohol ingestion impairs airway epithelial integrity and renders individuals susceptible to lung injury.
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影响因子：
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作者：
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