多房棘球蚴诱导小鼠枯否氏细胞极化相关ceRNA调控网络的构建与分析

doi:10.11843/j.issn.0366-6964.2022.12.026

摘要/Abstract

摘要： 多房棘球蚴(Em)是一种危害严重的人畜共患寄生虫。为探索非编码RNA在多房棘球蚴感染诱导小鼠肝枯否氏细胞(KCs)极化中的作用机制，本研究用多房棘球蚴原头蚴感染BALB/c小鼠，分离感染2月小鼠肝KCs，通过RNA测序分析，构建与多房棘球蚴感染引起的KCs M2型极化相关的竞争性内源RNA (competing endogenous RNAs,ceRNA)调控网络，并通过qPCR和miRNA过表达进行验证。结果表明，多房棘球蚴感染小鼠2月后，KCs M1型标志分子均显著下调，而M2型标志分子均显著上调。5个circRNA和9个miRNA参与M2型标志分子IL10、IL13、Arg1基因的表达调控，而且其表达趋势与测序结果及circRNA-miRNA-mRNA调节关系基本一致。另外，miR-466c-5p过表达导致了circ_0000372和IL13的表达下调，说明可能存在circ_0000372-miR-466c-5p-IL13轴的调节关系，并在多房棘球蚴感染诱导小鼠肝KCs极化中发挥重要作用。本研究为进一步揭示多房棘球蚴调控宿主巨噬细胞极化的相关机制提供了重要理论依据。

关键词: 多房棘球蚴, M2型巨噬细胞极化, ceRNA, 调控网络

Abstract: Echinococcus multilocularis is a severe zoonotic parasite. To explore the mechanism of non-coding RNA in the polarization of mice liver Kupffer cells (KCs) induced by E. multilocularis infection, we herein infected the BALB/c mouse with E. multilocularis and isolated the KCs from the mice liver at 2 months post infection (Mpi). After RNA high-throughput sequencing, we analyzed and constructed the ceRNA regulatory network related to the M2 polarization of KCs induced by E. multilocularis infection. The qPCR and miRNA overexpression assays were used to verify the ceRNA regulatory network. The results showed that, after E. multilocularis infection 2 Mpi, the M1-type markers were significantly down-regulated, while the M2-type markers were significantly up-regulated. Among them, 5 circRNAs and 9 miRNAs were identified to be involved in regulating the expression of M2-type markers, such as IL10, IL13, and Arg1, and their expression patterns were consistent with the sequencing results and the circRNA-miRNA-mRNA regulatory relationship. Moreover, overexpression of miR-466c-5p led to the down-regulation of circ_0000372 and IL13, indicating that the circ_0000372-miR-466c-5p-IL13 axis plays vital role in the polarization of mouse liver macrophages induced by E. multilocular infection. This study provides an essential theoretical basis for further revealing the mechanism of E. multilocularis regulating the polarization of host macrophages.

Key words: Echinococcus multilocularis, polarization of M2-type macrophages, ceRNA, regulation networks

中图分类号:

S852.734

李艳萍, 刘婷丽, 李红, 陈国梁, 王立群, 郭小腊, 骆学农. 多房棘球蚴诱导小鼠枯否氏细胞极化相关ceRNA调控网络的构建与分析[J]. 畜牧兽医学报, 2022, 53(12): 4410-4418.

LI Yanping, LIU Tingli, LI Hong, CHEN Guoliang, WANG Liqun, GUO Xiaola, LUO Xuenong. Construction and Analysis of Polarization-related ceRNA Regulatory Network in Mouse Kupffer Cells Induced by Echinococcus multilocularis[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4410-4418.

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