SerpinE1通过JAK/STAT通路调节HIV-1在巨噬细胞中的复制

SerpinE1 regulating HIV-1 replication in macrophages through the JAK/STAT pathway

陆贝贝 ¹陈姗姗 ¹陈飞蓉 ¹石敏娟 ¹吴玉婷 ¹叶力 ¹梁浩 ¹苏锦明 ¹蒋俊俊¹

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作者信息

1. 广西医科大学公共卫生学院广西艾滋病防治研究重点实验室,南宁 530021
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摘要

目的:探讨丝氨酸蛋白酶抑制剂E家族成员1(SerpinE1)与人类免疫缺陷病毒1型(HIV-1)感染的关系,及其作为一种蛋白酶抑制剂在巨噬细胞中对HIV感染过程发挥的作用和机制.方法:按年龄、性别特征成组匹配,招募HIV感染未治疗人群[HIVART(-)组]和健康对照人群(HC组),并检测外周血单个核细胞(PBMCs)中SerpinE1 mRNA表达量.在THP-1细胞中构建SerpinE1敲低细胞(敲低SerpinE1组),感染或不感染HIV BaL.酶联免疫吸附试验(ELISA)法检测HIV-p24和干扰素(IFN)-α蛋白水平,有参转录组测序分析染毒后敲低SerpinE1组与对照组的差异基因和KEGG富集通路,实时荧光定量PCR(RT-qPCR)法检测HIV-1 Gag、Toll样受体7(TLR7)、Toll样受体8(TLR8)、白细胞介素-1β(IL-1β)、MX动力蛋白样GT-Pase 1(MX1)、MX 动力蛋白样 GTPase 2(MX2)mRNA 相对表达量,western blotting 法检测 JAK2、STAT1、STAT2、SATA4、IL-1β和MX1蛋白表达水平.结果:与HC组比较,HIVART(-)组SerpinE1表达水平降低,并且在THP-1来源的巨噬细胞中能够被HIV诱导下调(P＜0.05);敲低SerpinE1表达下调HIV-p24蛋白表达和HIV Gag mRNA表达,促进IFN-α蛋白分泌水平,促进TLR7、TLR8、Janus激酶2(JAK2)、信号转导子和转录激活子(STAT)蛋白家族的STAT1、STAT2、SATA4及IL-1β、MX1、MX2的表达(P＜0.05).结论:SerpinE1可能通过抑制TLR7和TLR8信号通路的激活,减少IFN-α的释放,进而抑制JAK/STAT通路及其诱导的多种IFN刺激基因和IFN相关因子表达,从而促进了 HIV-1的感染复制.

Abstract

Objective:To explore the relationship between serine protease inhibitor E family member 1(Ser-pinE1)and human immunodeficiency virus type 1(HIV-1)infection,as well as the role and mechanism of Ser-pinE1 as a protease inhibitor in macrophages in the process of HIV infection.Methods:Groups were matched ac-cording to age and gender characteristics to recruit untreated HIV infected patients[HIV ART(-)group]and healthy controls(HC group),and the expression of SerpinE1 mRNA in peripheral blood mononuclear cells(PBMCs)was detected.SerpinE1 knockdown cells were constructed on THP-1 cells(knock-down SerpinE1 group),infected or uninfected with HIV BaL.The expression levels of HIV-p24 and interferon(IFN)-α protein were detected by enzyme-linked immunosorbent assay(ELISA).The differential genes and KEGG enrichment pathways between knock-down SerpinE1 group and control group after exposure were analyzed by transcriptome sequencing.The mRNA expression levels of HIV Gag,Toll-like receptor 7(TLR7),Toll-like receptor 8(TLR8),interleukin-1β(IL-1β),MX dynamin-like GTPase 1(MX1)and MX2 were detected by reverse transcription-quan-titative PCR(RT-qPCR).The protein expression levels of JAK2,STAT1,STAT2,SATA4,IL-1β and MX1 were detected by western blotting.Results:Compared with the HC group,SerpinE1 was lowly expressed in in the HIV ART(-)group,and could be down-regulated by HIV in THP-1-derived macrophages(P＜0.05).Knockdown of SerpinE1 expression down-regulated HIV-p24 protein expression and HIV Gag mRAN expression,promoted IFN-α protein secretion level,and enhanced the expression of TLR7,TLR8,Janus kinase 2(JAK2),signal trans-ducer and activator of transcription(STAT)protein family members STAT1,STAT2,STAT4,IL-1β,MX1,and MX2(P＜0.05).Conclusion:SerpinE1 may reduce the release of IFN by inhibiting the activation of TLR7 and TLR8 signaling pathways,and then inhibit the JAK/STAT pathway and its induced expression of various IFN stimulated genes and IFN-related factors,thereby promoting HIV-1 infection and replication.

关键词

丝氨酸蛋白酶抑制剂E家族成员1/巨噬细胞/干扰素-α/JAK/STAT/抗人类免疫缺陷病毒1型

Key words

serine protease inhibitor E family member 1/macrophages/interferon-alpha/JAK/STAT/anti-hu-man immunodeficiency virus type 1

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基金项目

国家自然科学基金资助项目(82373640)

国家自然科学基金资助项目(82103898)

出版年

2024

广西医科大学学报

广西医科大学

广西医科大学学报

CSTPCD

影响因子：0.788

ISSN：1005-930X

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