gga-miR-142-3p negatively regulates Mycoplasma gallisepticum (HS strain)-induced inflammatory cytokine production via the NF-κB and MAPK signaling by targeting TAB2
gga-miR-142-3p negatively regulates Mycoplasma gallisepticum (HS strain)-induced inflammatory cytokine production via the NF-κB and MAPK signaling by targeting TAB2
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GgA-miR-142-3p通过靶向TAB2，通过NF-κB和MAPK信号负性调节鸡败血支原体(HS株)诱导炎性细胞因子的产生
DOI：
10.1007/s00011-021-01499-2
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发表时间：
2021-09-23
影响因子：
6.7
通讯作者：
Peng, Xiuli
中科院分区：
文献类型：
--
作者：
Yang, Yaping;Wang, Yingjie;Peng, Xiuli
目的 鸡毒支原体（MG）是一种臭名昭著的禽类病原体，给养禽业带来了巨大的经济损失。 MG 感染的特点是严重、无法控制的炎症和宿主 DNA 损伤。微核糖核酸（miRNA）已成为微生物发病机制中的重要调节因子。然而，miRNA 在 MG 感染中的作用尚不清楚。在本研究中，我们验证了gga-miR-142-3p的功能作用。方法通过逆转录定量实时PCR分析确定gga-miR-142-3p在MG感染的鸡胚胎和MG感染的鸡胚胎成纤维细胞系（DF-1）的肺部中的相对表达量。利用生物信息学数据库分析gga-miR-142-3p的靶基因。进行荧光素酶报告基因测定以及基因表达分析以验证靶基因。为了进一步探讨 MG 感染后 gga-miR-142-3p 的生物学功能，使用 Cell Counting Kit-8 (CCK-8) 定量细胞增殖。同时，使用流式细胞仪测量细胞周期分析和细胞凋亡。结果gga-miR-142-3p在MG感染的鸡胚肺和DF-1细胞中均显着上调。 MG感染后gga-miR-142-3p过表达显着下调促炎细胞因子的表达，包括白细胞介素1β、白细胞介素6和肿瘤坏死因子α。同时，gga-miR-142-3p通过促进细胞增殖、促进细胞进展和抑制细胞凋亡来增强宿主对MG感染的防御。有趣的是，TAB2敲低组显示出相似的结果，而TAB2过表达组和gga-miR-142-3p抑制剂组则具有完全相反的结果。当 gga-miR-142-3p 过表达时，核因子 kappa B (NF-kappa B) 中 p-p65 和丝裂原激活蛋白激酶 (MAPK) 通路中 p-p38 的表达降低。结论 MG 感染后，gga-miR-142-3p 的上调通过靶向 TAB2 负向调节 NF-kappa B 和 MAPK 信号通路来减轻炎症，并促进通过抑制细胞凋亡和促进细胞周期进程来抑制细胞增殖，从而防御 MG 感染。
Objective Mycoplasma gallisepticum (MG), a notorious avian pathogen, leads to considerable economic losses in the poultry industry. MG infection is characterized by severe, uncontrollable inflammation and host DNA damage. Micro ribonucleic acids (miRNAs) have emerged as important regulators in microbial pathogenesis. However, the role of miRNAs in MG infection is poorly characterized. In this study, we validated the functional roles of gga-miR-142-3p.Methods The relative expression of gga-miR-142-3p in the lungs of the MG-infected chicken embryos and the MG-infected chicken embryonic fibroblast cell line (DF-1) was determined by reverse transcription quantitative real-time PCR analysis. Bioinformatics database was used to analysis the target gene of gga-miR-142-3p. The luciferase reporter assay as well as gene expression analysis were conducted to validate the target gene. To further explore the biological functions of gga-miR-142-3p upon MG infection, the cell proliferation was quantified using Cell Counting Kit-8 (CCK-8). Meanwhile, cell cycle analysis and apoptosis were measured using a flow cytometer.Results gga-miR-142-3p was significantly upregulated in both MG-infected chicken-embryo lungs and the DF-1 cells. gga-miR-142-3p over expression significantly downregulated the expression of pro-inflammatory cytokines, including interleukin-1 beta, interleukin-6 and tumor necrosis factor alpha after MG infection. Meanwhile, gga-miR-142-3p enhanced the host defense against MG infection by facilitating cell proliferation, promoting cell progression and inhibiting cell apoptosis. Interestingly, TAB2 knockdown groups show similar results, whereas, TAB2 over-expression groups and gga-miR-142-3p inhibitor groups had thoroughly opposite results. The expression of p-p65 in nuclear factor kappa B (NF-kappa B) and p-p38 in the mitogen-activated protein kinase (MAPK) pathway was decreased when gga-miR-142-3p was over-expressed.Conclusion Upon MG infection, upregulation of gga-miR-142-3p alleviates inflammation by negatively regulating the signaling pathways of NF-kappa B and MAPKs by targeting TAB2 and facilitates cell proliferation by inhibiting cell apoptosis and promoting cell cycle progression to defend against MG infection.