Exosomal miR-181a-5p reduce Mycoplasma gallisepticum (HS strain) infection in chicken by targeting PPM1B and activating the TLR2-mediated MyD88/NF-κB signaling pathway
Exosomal miR-181a-5p reduce Mycoplasma gallisepticum (HS strain) infection in chicken by targeting PPM1B and activating the TLR2-mediated MyD88/NF-κB signaling pathway
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DOI：
10.1016/j.molimm.2021.09.005
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发表时间：
2021-10-26
影响因子：
3.6
通讯作者：
Peng, Xiuli
中科院分区：
文献类型：
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作者：
Sun, Yingfei;Wang, Yingjie;Peng, Xiuli
鸡毒支原体（MG）是引起鸡慢性呼吸道疾病（CRD）的重要病原之一。已经充分证明从细胞分泌的外泌体将miRNA递送至受体细胞以调节细胞功能。本研究旨在探讨外泌体miR-181 a-5 p在MG-HS感染中的作用及其机制。在本研究中，我们发现，在体内和体外的miR-181 a-5 p表达显著上调MG-HS感染后。它也在来源于MG-HS感染的II型肺细胞（CP-II）的外来体中上调。此外，由MG-HS感染的CP-II分泌的外泌体能够将miR-181 a-5 p转移至受体鸡胚成纤维细胞（DF-1），导致受体DF-1细胞中miR-181 a-5 p表达显著上调。我们进一步鉴定了Mg 2 +/Mn 2+依赖性蛋白磷酸酶1B（PPM 1B）是miR-181 a-5 p的靶基因。过表达miR-181 a-5 p或敲低PPM 1 B激活核因子-κ B（NF-κ B B）信号通路，而抑制miR-181 a-5 p和过表达PPM 1 B则导致相反的结果。此外，上调miR-181 a-5 p可显著增加Toll样受体2（TLR 2）、髓样分化因子88（MyD 88）、肿瘤坏死因子α（TNF-α）和白细胞介素-1 β（IL-1 β）的表达，而抑制miR-181 a-5 p则显示相反的结果。上调miR-181 a-5 p可促进细胞增殖、细胞周期进程，抑制细胞凋亡，从而抵抗MG-HS感染。miR-181 a-5 p的过表达通过直接抑制PPM 1B而显著负调控鸡毒支原体粘附素蛋白（pMGA 1. 2）的表达。因此，我们认为来源于CP-II细胞的exosomal miR-181 a-5 p通过直接靶向PPM 1 B，激活TLR 2介导的MyD 88/NF-κ B B信号通路，促进促炎细胞因子的表达，从而防御受体DF-1细胞的MG-HS感染。
Mycoplasma gallisepticum (MG) is one of the most important pathogens that causes chronic respiratory disease (CRD) in chickens. Exosomes secreted from cells have been well demonstrated to deliver miRNAs to recipient cells to modulate cellular functions. The purpose of this study is to explore the underlying functions and mechanisms of exosomal miR-181a-5p in MG-HS infection. In this study, we found that miR-181a-5p expression in vivo and in vitro was significantly up-regulated after MG-HS infection. It was also upregulated in exosomes, which were derived from MG-HS-infected type-II pneumocytes cells (CP-II). In addition, exosomes secreted by MG-HS-infected CP-II were able to transfer miR-181a-5p to recipient chicken embryo fibroblast cells (DF-1), resulting in a significant upregulation of miR-181a-5p expression in recipient DF-1 cells. We further identified that Mg2+/Mn2+-dependent protein phosphatase 1B (PPM1B) was the target gene of miR-181a-5p. Overexpression of miR-181a-5p or knockdown of PPM1B activated the nuclear factor-kappa B (NF-kappa B) signaling pathway, whereas inhibition of miR-181a-5p and overexpression of PPM1B led to the opposite results. Besides, upregulation of miR-181a-5p significantly increased the expression of toll-like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88), tumor necrosis factors alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta), whereas inhibition of miR-181a-5p showed a contrary result. Up-regulation of miR-181a-5p promoted cell proliferation, cell cycle progression and inhibited apoptosis to resist MG-HS infection. Moreover, overexpression of miR-181a-5p significantly negative regulated the expression of Mycoplasma gallisepticum adhesin protein (pMGA1.2) by directly inhibiting PPM1B. Thus, we concluded that exosomal miR-181a-5p from CP-II cells activated the TLR2mediated MyD88/NF-kappa B signaling pathways by directly targeting PPM1B to promote the expression of proinflammatory cytokines for defending against MG-HS infection in recipient DF-1 cells.