乌头酸脱羧酶1对BCG诱导巨噬细胞炎症反应的调控作用研究

doi:10.11843/j.issn.0366-6964.2024.04.031

摘要/Abstract

摘要： 旨在探究乌头酸脱羧酶1(aconitate decarboxylase 1，ACOD1)对减毒牛分枝杆菌卡介苗(Bacillus Calmette-Guérin，BCG)诱导的小鼠巨噬细胞RAW264.7炎症反应的调控作用。基于小干扰RNA技术构建ACOD1敲减模型，采用Western blot、qRT-PCR、免疫荧光等技术检测BCG感染巨噬细胞RAW264.7后细胞内ACOD1、细胞因子IL-1β、TNF-α、IL-4、IL-6、IL-10、COX2、iNOS以及TLR4/MyD88/NF-κB信号通路蛋白的表达变化。结果显示，BCG感染巨噬细胞RAW264.7后以时间和浓度依赖性方式显著上调ACOD1及促炎细胞因子TNF-α、IL-1β、IL-6、COX2、iNOS的表达，下调抑炎细胞因子IL-4、TGF-β的表达(P < 0.01)；si-ACOD1结合BCG感染会显著下调促炎细胞因子表达(P < 0.01)，同时也会下调TLR4/MyD88/NF-κB通路相关蛋白TLR4、MyD88、TRAF6、p-NF-κB p65的表达。综上, ACOD1可能通过TLR4/MyD88/NF-κB信号通路参与调控BCG引起的炎症反应。

关键词: 乌头酸脱羧酶1, BCG, 小鼠巨噬细胞RAW264.7, 炎症反应, TLR4/MyD88/NF-κB信号通路

Abstract: Tuberculosis (TB) is a chronic respiratory disease caused by Mycobacterium tuberculosis (Mtb). There is mounting evidence that inflammatory response plays an important role in regulating the host immune responses. Aconitate decarboxylase 1 (ACOD1) is a stress-related inducible protein associated with inflammation and infection, many studies have identified ACOD1 as one of the most upregulated genes under various conditions associated with infection. However, the role of ACOD1 in the regulation of RAW264.7 macrophage inflammatory response induced by attenuated Mycobacterium bovis Bacillus Calmette-Guérin (BCG) infection remains unclear. In this study, we investigated the effect of ACOD1 on RAW264.7 cell inflammatory response during BCG infection. In addition, we explored the role of ACOD1 in regulating BCG-induced RAW264.7 cell inflammatory response using small interfering RNAs targeting ACOD1. The experiment used Western blot, Quantitative Real-time PCR (qRT-PCR) and immunofluorescence to detect the levels of ACOD1, cytokines and TLR4/MyD88/NF-κB signaling pathway proteins. The results demonstrated that BCG could induce macrophage inflammatory response and ACOD1 upregulation in a time and concentration-dependent manner; Knockdown of ACOD1 could attenuate BCG-induced inflammatory response in RAW264.7 cells. Moreover, we found that ACOD1 knockdown decreased the expression of IL-1β, TNF-α, IL-6, COX2, iNOS, and increased the levels of anti-inflammatory cytokines about IL-4 and IL-10; ACOD1 can also activate TLR4/MyD88/NF-κB signaling pathway, decreased the levels of TLR4, MyD88, TRAF6, p-NF-κB p65. These results indicated that ACOD1 could regulate inflammatory response caused by BCG through TLR4/MyD88/NF-κB signaling pathway.

Key words: aconitate decarboxylase 1, BCG, RAW264.7 cell, inflammatory response, TLR4/MyD88/NF-κB signaling pathway

中图分类号:

S852.618

戴帆, 刘占有, 张旭阳, 李武. 乌头酸脱羧酶1对BCG诱导巨噬细胞炎症反应的调控作用研究[J]. 畜牧兽医学报, 2024, 55(4): 1696-1706.

DAI Fan, LIU Zhanyou, ZHANG Xuyang, LI Wu. Aconitate Decarboxylase 1 Could Regulate the Inflammatory Response Caused by BCG[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1696-1706.

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