House dust mite extract activates EphA2-STAT3/p38 MAPK signaling pathway in airway epithelial cells to mediate airway inflammation].

Guoping Huang, Qing Yuan, Yingchun Ye, Hong Yu, Xiang Xie, Wenfeng Xu, Siji Nian

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 2021 Jan

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Objective To investigate the effect of ephrin type-A receptor 2 (EphA2) on the expression of inflammatory cytokines in airway epithelial cells induced by house dust mite extract (HDM) and the underlying mechanism. Methods The cell model of EphA2 knockdown was established by transfection of EphA2 siRNA into airway cell line 16HBE cells. After the 16HBE cells were stimulated with HDM, the mRNA levels of EphA2, interleukin 6 (IL-6) and IL-8 were determined by real-time quantitative PCR (qPCR), and the protein levels of IL-6, IL-8, IL-17A, IL-17F and tumor necrosis factor-α (TNF-α) were measured by cytometric bead array (CBA). Western blotting was used to analyze the protein expression of EphA2, phosphorylated EphA2 (p-EphA2), signal transducer and activator of transcription (STAT3), phosphorylated STAT3 (p-STAT3), p38 mitogen-activated protein kinases (p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), nuclear factor κ-B p65 (NF-κB p65) and phosphorylated NF-κB p65 (p-NF-κB p65). Then, in the 16HEB cells stimulated by STAT3 inhibitor Stattic or p38 MAPK inhibitor SB203580 in combination with HDM, the mRNA and protein expression levels of IL-6 and IL-8 were detected by qPCR and CBA. Results Knockdown of EphA2 significantly inhibited the expression of IL-6 and IL-8 in HDM-induced 16HBE, and reduced the total protein and phosphorylated levels of STAT3 and p38 MAPK, but had no significant effect on the total protein and phosphorylated levels of NF-κB p65. After stattic inhibited the expression and activation of STAT3, the mRNA and protein levels of IL-6 and IL-8 significantly decreased in HDM-induced 16HBE cells. Interestingly, while SB203580 inhibited the activation of p38 MAPK signaling pathway, it only inhibited the mRNA levels of IL-6 and IL-8 in HDM-induced 16HBE cells, but had no effect on their protein levels. Conclusion HDM can induce the expression of IL-6 and IL-8 in 16HBE cells to participate in airway inflammation by activating the EphA2-STAT3/p38 MAPK pathway.

Citation

Guoping Huang, Qing Yuan, Yingchun Ye, Hong Yu, Xiang Xie, Wenfeng Xu, Siji Nian. House dust mite extract activates EphA2-STAT3/p38 MAPK signaling pathway in airway epithelial cells to mediate airway inflammation]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 2021 Jan;37(1):31-38