Mesenchymal Stem Cell-Conditioned Medium Rescues LPS-Impaired ENaC Activity in Mouse Trachea via WNK4 Pathway
Mesenchymal Stem Cell-Conditioned Medium Rescues LPS-Impaired ENaC Activity in Mouse Trachea via WNK4 Pathway
复制标题
间充质干细胞条件培养基通过 WNK4 途径挽救小鼠气管中 LPS 受损的 ENaC 活性
DOI：
10.2174/1381612826666200131141732
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发表时间：
2020-01-01
影响因子：
3.1
通讯作者：
Nie, Hongguang
中科院分区：
文献类型：
--
作者：
Hou, Yapeng;Zhou, Zhiyu;Nie, Hongguang
背景：气道上皮作为宿主防御的第一道防线，在维持呼吸系统的稳态和功能中发挥着重要作用。值得注意的是，上皮钠通道 (ENaC) 是 LPS 诱导的气道损伤的受害者之一。鉴于间充质干细胞（MSCs）在气道损伤再生医学领域的巨大前景以及基于细胞的MSCs治疗的局限性，我们重点研究了间充质干细胞条件培养基（MSCs-CM）对小鼠气管上皮细胞ENaC活性的治疗作用。方法：采用小室装置记录原代培养的小鼠气管上皮细胞的短路电流，以反映ENaC活性。 a 和 ? 的表达式分别通过蛋白质印迹和实时 PCR 测量 ENaC 的蛋白质和 mRNA 水平。在蛋白水平上测量with-no-lysinekinase-4（WNK4）和ERK1/2的表达，并通过敲低WNK4来确定WNK4和ERK1/2之间的关系。结果：MSCs-CM恢复了LPS受损的ENaC活性，并增强了原代培养的小鼠气管上皮细胞中ENaC的mRNA和蛋白表达。同时，在LPS诱导的气道损伤中，给予MSCs-CM后，ENaC的负调节因子WNK4和ERK1/2相应受到抑制。 siRNA敲除WNK4基因后，ERK1/2磷酸化水平下降。结论：鉴于ENaC在液体重吸收中的关键作用以及MSCs-CM在气道上皮损伤中的有益作用，我们的研究结果表明MSCs-CM可通过WNK4-ERK1/2途径有效缓解LPS诱导的ENaC功能障碍，这将为气道损伤的治疗提供有效的方向。
Background: Airway epithelium plays an essential role in maintaining the homeostasis and function of respiratory system as the first line of host defense. Of note, epithelial sodium channel (ENaC) is one of the victims of LPS-induced airway injury. Regarding the great promise held by mesenchymal stem cells (MSCs) for regenerative medicine in the field of airway injury and the limitations of cell-based MSCs therapy, we focused on the therapeutic effect of MSCs conditioned medium (MSCs-CM) on the ENaC activity in mouse tracheal epithelial cells.Methods: Ussing chamber apparatus was applied to record the short-circuit currents in primary cultured mouse tracheal epithelial cells, which reflects the ENaC activity. Expressions of a and ? ENaC were measured at the protein and mRNA levels by western blot and real-time PCR, respectively. The expression of with-no-lysinekinase- 4 (WNK4) and ERK1/2 were measured at protein levels, and the relationship between WNK4 and ERK1/2 was determined by WNK4 knockdown.Results: MSCs-CM restored the LPS-impaired ENaC activity, as well as enhanced the mRNA and protein expressions of ENaC in primary cultured mouse tracheal epithelial cells. Meanwhile, WNK4 and ERK1/2, both negative-regulators of ENaC, were suppressed accordingly after the administration of MSCs-CM in LPS-induced airway injury. After WNK4 gene was knocked down by siRNA, the level of ERK1/2 phosphorylation decreased.Conclusion: In light of the key role of ENaC in fluid reabsorption and the beneficial effects of MSCs-CM in the injury of airway epithelium, our results suggest that MSCs-CM is effective in alleviating LPS-induced ENaC dysfunction through WNK4-ERK1/2 pathway, which will provide a potent direction for the therapy of airway injury.