mmu-miR-374b-5p 通过下调库普弗细胞中 C/EBPβ/NF-κB 信号通路调节多房棘球蚴感染期间的炎症因子。

mmu-miR-374b-5p modulated inflammatory factors via downregulation of C/EBP β/NF-κB signaling in Kupffer cells during Echinococcus multilocularis infection.

机构信息

State Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou, 730046, Gansu Province, People's Republic of China.

Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, 225009, People's Republic of China.

出版信息

Parasit Vectors. 2024 Mar 29;17(1):163. doi: 10.1186/s13071-024-06238-0.

DOI:10.1186/s13071-024-06238-0

PMID:38553755

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10981327/

Abstract

BACKGROUND

Alveolar echinococcosis (AE) is an important infectious disease caused by the metacestode larvae of Echinococcus multilocularis, seriously threatening global public health security. Kupffer cells (KCs) play important roles in liver inflammatory response. However, their role in hepatic alveolar echinococcosis has not yet been fully elucidated.

METHODS

In this study, qRT-PCR was used to detect the expression level of miR-374b-5p in KCs. The target gene of miR-374b-5p was identified through luciferase reporter assays and loss of function and gains. Critical genes involved in NFκB signaling pathway were analyzed by qRT-PCR and western blot.

RESULTS

This study reported that miR-374b-5p was significantly upregulated in KCs during E. multilocularis infection and further showed that miR-374b-5p was able to bind to the 3'-UTR of the C/EBP β gene and suppressed its expression. The expression levels of NF-κBp65, p-NF-κBp65 and pro-inflammatory factors including iNOS, TNFα and IL6 were attenuated after overexpression of miR-374b-5p while enhanced after suppression of miR-374b-5p. However, the Arg1 expression level was promoted after overexpression of miR-374b-5p while suppressed after downregulation of miR-374b-5p. Additionally, increased protein levels of NF-κBp65 and p-NF-κBp65 were found in the C/EBP β-overexpressed KCs.

CONCLUSIONS

These results demonstrated that miR-374b-5p probably regulated the expression of inflammatory factors via C/EBP β/NF-κB signaling. This finding is helpful to explore the mechanism of inflammation regulation during E. multilocularis infection.

摘要

背景

泡型包虫病（AE）是一种由细粒棘球绦虫的幼虫引起的重要传染病，严重威胁着全球公共卫生安全。枯否细胞（KCs）在肝脏炎症反应中发挥重要作用。然而，它们在肝泡型包虫病中的作用尚未完全阐明。

方法

本研究采用 qRT-PCR 检测 KCs 中 miR-374b-5p 的表达水平。通过荧光素酶报告基因检测和功能丧失和获得实验鉴定 miR-374b-5p 的靶基因。采用 qRT-PCR 和 Western blot 分析 NFκB 信号通路中涉及的关键基因。

结果

本研究报道 miR-374b-5p 在泡型包虫感染过程中在 KCs 中显著上调，并进一步表明 miR-374b-5p 能够与 C/EBPβ 基因的 3'-UTR 结合并抑制其表达。miR-374b-5p 过表达后，NF-κBp65、p-NF-κBp65 和促炎因子（包括 iNOS、TNFα 和 IL6）的表达水平降低，而抑制 miR-374b-5p 后表达水平增强。然而，miR-374b-5p 过表达后 Arg1 表达水平升高，而抑制 miR-374b-5p 后 Arg1 表达水平降低。此外，在 C/EBPβ 过表达的 KCs 中发现 NF-κBp65 和 p-NF-κBp65 的蛋白水平增加。