Knockdown of ITIH4 reduces inflammatory damage and apoptosis of A549 cells induced by Mycoplasma pneumoniae through NLRP3 inflammation.

IF 2.1 4区医学 Q3 ALLERGY

Allergologia et immunopathologia Pub Date : 2025-07-01 eCollection Date: 2025-01-01 DOI:10.15586/aei.v53i4.1367

Zhinan Zhang, Yixian Zhang, Bihe Zeng

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引用次数: 0

Abstract

Background: Mycoplasma pneumoniae (MP) is a leading cause of community-acquired respiratory infections in pediatric patients. This study aimed to investigate whether the pro-inflammatory function of inter-alpha-trypsin inhibitor heavy chain (ITIH4) contributes to the pathogenesis of MP-induced pneumonia.

Method: A549 cells were stimulated with MP to model pneumonia in vitro. ITIH4 expression was knocked down in A549 cells using lentiviral transfection. Cell viability was measured using the Cell Counting Kit-8 (CCK-8) assay, while cell apoptosis was assessed via flow cytometry. The concentrations of pro-inflammatory (IL-6, TNF-α) and anti-inflammatory (IL-10) cytokines were quantified using enzyme-linked immunosorbent assay (ELISA). Western blotting was conducted to detect apoptosis-related proteins and components of the NLRP3 inflammasome.

Result: MP stimulation led to increased ITIH4 expression in A549 cells, and knockdown of ITIH4 prevented the MP-induced reduction in cell viability. Moreover, ITIH4 knockdown reduced the release of inflammatory cytokines in response to MP and significantly decreased MP-induced apoptosis. In addition, ITIH4 knockdown inhibited activation of the NLRP3 inflammasome, while reactivation of NLRP3 reversed the protective effects associated with ITIH4 knockdown.

Conclusion: ITIH4 knockdown alleviates MP-induced inflammatory damage and cell death in A549 cells by inhibiting activation of the NLRP3 inflammasome.

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敲低ITIH4可通过NLRP3炎症降低肺炎支原体诱导的A549细胞的炎症损伤和凋亡。

背景：肺炎支原体（MP）是儿科患者社区获得性呼吸道感染的主要原因。本研究旨在探讨α -胰蛋白酶抑制剂间重链（ITIH4）的促炎功能是否参与mp诱导肺炎的发病机制。方法：用MP刺激A549细胞制备肺炎模型。采用慢病毒转染法降低A549细胞中ITIH4的表达。采用细胞计数试剂盒-8 （CCK-8）检测细胞活力，流式细胞术检测细胞凋亡。采用酶联免疫吸附法（ELISA）测定促炎因子（IL-6、TNF-α）和抗炎因子（IL-10）的浓度。Western blotting检测NLRP3炎性小体的凋亡相关蛋白和组分。结果：MP刺激导致A549细胞中ITIH4表达增加，而抑制ITIH4可阻止MP诱导的细胞活力降低。此外，ITIH4敲低可减少MP反应中炎症因子的释放，并显著减少MP诱导的细胞凋亡。此外，ITIH4敲低抑制了NLRP3炎性体的激活，而NLRP3的再激活逆转了与ITIH4敲低相关的保护作用。结论：ITIH4敲低可通过抑制NLRP3炎性小体的激活，减轻mp诱导的A549细胞炎症损伤和细胞死亡。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Allergologia et immunopathologia 医学-过敏

CiteScore

3.70

自引率

0.00%

发文量

131

审稿时长

6-12 weeks

期刊介绍： Founded in 1972 by Professor A. Oehling, Allergologia et Immunopathologia is a forum for those working in the field of pediatric asthma, allergy and immunology. Manuscripts related to clinical, epidemiological and experimental allergy and immunopathology related to childhood will be considered for publication. Allergologia et Immunopathologia is the official journal of the Spanish Society of Pediatric Allergy and Clinical Immunology (SEICAP) and also of the Latin American Society of Immunodeficiencies (LASID). It has and independent international Editorial Committee which submits received papers for peer-reviewing by international experts. The journal accepts original and review articles from all over the world, together with consensus statements from the aforementioned societies. Occasionally, the opinion of an expert on a burning topic is published in the "Point of View" section. Letters to the Editor on previously published papers are welcomed. Allergologia et Immunopathologia publishes 6 issues per year and is included in the major databases such as Pubmed, Scopus, Web of Knowledge, etc.