作者： Wu, Xiaoxia;Li, Kaiming;Xie, Meihua;Yu, Minjun;Tang, Shuangyang;...

期刊： MOLECULAR MEDICINE REPORTS,2018年17(2):3178-3185 ISSN：1791-2997

通讯作者： Hu, Sihai

作者机构： [Wu, Xiaoxia; Hu, Sihai; Li, Zhenyu; Li, Kaiming; Xie, Meihua; Yu, Minjun; Tang, Shuangyang] Univ South China, Lab Antiinfect Immun, Pathogen Biol Inst, Coll Basic Med, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Li, Kaiming] Cent Hosp Hengyang City, Clin Lab, Hengyang 421001, Hunan, Peoples R China.

通讯机构： [Hu, Sihai] U;Univ South China, Lab Antiinfect Immun, Pathogen Biol Inst, Coll Basic Med, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

关键词： CpG oligodeoxynucleotide;DNA vaccine;gamma interferon;genomic DNA;immunoglobulin G;immunoglobulin G1;immunoglobulin G2a;interleukin 4;Meningococcus vaccine;Neisseria meningitidis serogroup B outer membrane protein 0315;outer membrane protein;unclassified drug;bacterial antigen;bacterial protein;bacterium antibody;cytokine;DNA vaccine;immunoglobulin G;animal experiment;animal model;antibody titer;Article;bacterial strain;bactericidal activity;cellular immunity;comparative study;controlled study;COS-7 cell line;DNA immunization;eukaryotic cell;female;genetic transfection;humoral immunity;in vitro study;in vivo study;meningococcosis;mouse;Neisseria meningitidis;newborn;nonhuman;protein blood level;RAW 264.7 cell line;recombinant plasmid;survival rate;vaccine immunogenicity;animal;Bagg albino mouse;blood;Chlorocebus aethiops;CV-1 cell line;genetics;hamster;immunology;meningococcosis;metabolism;Neisseria meningitidis;pathogenicity;plasmid;serotype;veterinary;Animals;Antibodies, Bacterial;Antigens, Bacterial;Bacterial Proteins;Cercopithecus aethiops;COS Cells;Cricetinae;Cytokines;Female;Immunoglobulin G;Meningococcal Infections;Mice;Mice, Inbred BALB C;Neisseria meningitidis;Plasmids;RAW 264.7 Cells;Serogroup;Survival Rate;Vaccines, DNA

摘要： Neisseria meningitidis (N. meningitidis) is a major cause of meningitis and sepsis. Capsular polysaccharide-based vaccines against serogroups A, C, Y, and W135 are available; however, the development of a vaccine against N. meningitidis serogroup B (NMB) has been problematic. NMB0315 is an outer membrane protein of NMB that may be a virulence factor for N. meningitidis and a possible target for functional bactericidal antibodies. The present study aimed to develop a potent DNA vaccine against NMB by cloning the NMB0135 gene into the pcDNA3.1(+) vector to construct the recombinant plasmid pcDNA3.1(+)/NMB0315 (designated pNMB0315). pNMB0315 was transfected into eukaryotic COS-7 and RAW264.7 cells to express the recombinant (r)NMB0315 protein. Protective immunogenicity of the DNA vaccine was assessed in an in vivo mouse model. The levels of rNMB0315-specific immunoglobulin G (IgG), IgG1 and IgG2a antibodies in the pNMB0315-immunized group increased dramatically up to week 6 following the initial vaccination, and were significantly higher compared with the levels in the Control groups. The serum concentrations of interleukin-4 and interferon-γ were significantly higher in the pNMB0315-immunized group compared with the control groups. Following intraperitoneal challenge with a lethal dose of NMB strain MC58, the survival rate in the pNMB0315 + CpG group was 70% (14 out of 20 mice) at 14 days; by contrast, all mice in the control groups succumbed within 3 days. The serum bactericidal titers of the pNMB0315 + CpG group in vitro reached 1:128 following three immunizations. The results indicated that pNMB0315 may serve as a promising DNA vaccine against NMB. Copyright © 2017 Spandidos Publications. All rights reserved.

语种： 英文

作者： Zuo, Lingling;Sun, Hedong;Yu, Minjun;You, Xiaoxing;Zeng, Yanhua;...

期刊： Biotechnology & Biotechnological Equipment,2018年32(6):1590-1597 ISSN：1310-2818

通讯作者： Wu, Yimou

作者机构： [Zuo, Lingling] Univ South China, Affiliated Hosp 2, Dept Transfus Med, Hengyang, Peoples R China.;[Wu, Yimou; Zuo, Lingling; Yu, Minjun; You, Xiaoxing; Zeng, Yanhua] Univ South China, Hunan Prov Key Lab Special Pathogens Prevent & Co, Inst Pathogen Biol, Med Coll, Hengyang, Peoples R China.;[Wu, Yimou; Zuo, Lingling; Yu, Minjun; You, Xiaoxing; Zeng, Yanhua] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang, Peoples R China.;[Sun, Hedong] Univ South China, Dept Neurol, Affiliated Hosp 2, Hengyang, Peoples R China.

通讯机构： [Wu, Yimou] U;[Wu, Yimou] H;Univ South China, Hunan Prov Key Lab Special Pathogens Prevent & Co, Inst Pathogen Biol, Med Coll, Hengyang, Peoples R China.;Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang, Peoples R China.

关键词： Mycoplasma genitalium lipoproteins;LAMPs;tumour necrosis factor alpha (TNF-alpha);apoptosis;HeLa cells

摘要： This aim of this study was to investigate the effects of Mycoplasma genitalium lipoproteins on the tumour necrosis factor α (TNF-α)-induced apoptosis in HeLa cells. We separated the M. genitalium lipid-associated membrane proteins (LAMPs) from M. genitalium and used them to treat HeLa cells. Apoptosis was induced by TNF-α treatment. Cell cycle and apoptosis were detected by flow cytometry. Pro-inflammatory cytokine contents were determined by enzyme-linked immunosorbent assay (ELISA). Mitochondrial membrane potential and caspase-3 protease activity were also measured. There was no significant difference in the viable cell percentage in HeLa cells treated with M. genitalium LAMPs at 0–20 μg/mL. However, when treated with 40 μg/mL LAMPs, cytocidal activity was observed during the first 24 h. M. genitalium LAMPs induced cell cycle arrest at the G 1 phase in HeLa cells. When apoptosis was induced by TNF-α in HeLa cells, M. genitalium LAMPs significantly reduced the percentage of apoptotic cells. In addition, M. genitalium LAMPs significantly reduced the loss of mitochondrial membrane potential and caspase-3 protease activity in TNF-α-treated HeLa cells. However, the blocking of NF-κB significantly increased the mitochondrial membrane potential and caspase-3 protease activity in TNF-α-induced HeLa cells treated with M. genitalium LAMPs. The obtained results suggest that M. genitalium LAMPs could inhibit TNF-α-induced apoptosis in HeLa cells, which would contribute to the understanding of the pathogenesis of M. genitalium. © 2018, © 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

语种： 英文

作者： Deng, Xiangying;Zhu, Youcong;Dai, Pei;Yu, Minjun;Chen, Liesong;...

期刊： Microbial Pathogenesis,2018年120:140-146 ISSN：0882-4010

通讯作者： Zeng, Yanhua

作者机构： [Dai, Pei; Zhu, Youcong; Deng, Xiangying; Chen, Liesong; Zhu, Cuiming; Yu, Minjun; You, Xiaoxing; Zeng, Yanhua; Li, Lingling] Univ South China, Hunan Prov Key Lab Special Pathogens Prevent & Co, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pathogen Biol,Med Coll, Hengyang 421001, Peoples R China.

通讯机构： [Zeng, Yanhua] U;Univ South China, Hunan Prov Key Lab Special Pathogens Prevent & Co, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pathogen Biol,Med Coll, Hengyang 421001, Peoples R China.

关键词： adhesin;aspartyltryptophylserylseryltryptophylvalyltyrosylarginylaspartylprolylglutamylthreonine polypeptide;histidyltyrosylisoleucylaspartylphenylalanylarginyltryptophan polypeptide;polypeptide;protein antibody;recombinant protein;unclassified drug;valylhistidyltryptophylaspartylphenylalanylarginylglutamyltryptophyltrytophylglutamylprolylserine polypeptide;adhesin;bacterial protein;peptide;protein binding;Article;cell adhesion;cell interaction;controlled study;enzyme linked immunosorbent assay;host cell;human;human cell;immunofluorescence microscopy;in vitro study;Mycoplasma genitalium;nonhuman;peptide library;phage display;priority journal;protein binding;protein function;receptor binding;SV-HUC-1 cell line;antibody specificity;bacteriophage;bacterium adherence;cell line;chemistry;metabolism;Mycoplasma genitalium;mycoplasmosis;Adhesins, Bacterial;Antibody Specificity;Bacterial Adhesion;Bacterial Proteins;Bacteriophages;Cell Line;Enzyme-Linked Immunosorbent Assay;Humans;Mycoplasma genitalium;Mycoplasma Infections;Peptide Library;Peptides;Protein Binding;Recombinant Proteins

摘要： Mycoplasma genitalium adhesion protein (MgPa) is a major adhesin of M. genitalium, a human pathogen associated with a series of genitourinary tract diseases. MgPa plays a very important role in M. genitalium adhering to the host cells. However, the exact receptor peptides or proteins of MgPa are still poorly understood so far. Three polypeptides (V-H-W-D-F-R-Q-W-W-Q-P-S), (D-W-S-S-W-V -Y-R-D-P-Q-T) and (H-Y-I-D-F-R-W) were previously screened from a phage display random peptide library using recombinant MgPa (rMgPa) as a target molecule. In this study, three polypeptides were artificially synthesized and investigated as to whether they are potential receptors of MgPa. We found that rMgPa specifically bound to three synthesized polypeptides as determined via an indirect enzyme-linked immunosorbent assay (ELISA). Moreover, three polypeptides were further identified by indirect immunofluorescence microscopy (IFM). We confirmed that rMgPa and M. genitalium can adhere to SV-HUC-1 cells in vitro and that anti-rMgPa antibody and three synthesized polypeptides can partially inhibit the adherence of rMgPa and M. genitalium to SV-HUC-1 cells. In summary, these three polypeptides may be the essential receptor peptides of MgPa, and may aid in enhancing the understanding of biological function of MgPa and the possible pathogenic mechanism of M. genitalium. © 2018 Elsevier Ltd

语种： 英文

作者： 戴佩;邓湘赢;余敏君;李玲玲;罗丹;...

期刊： 中国免疫学杂志,2018年34(5):653-657 ISSN：1000-484X

作者机构： 南华大学病原生物学研究所, 特殊病原体防控湖南省重点实验室;[戴佩; 邓湘赢; 余敏君; 李玲玲; 罗丹; 曾焱华] 湖南省分子靶标新药研究协同创新中心, 衡阳, 421001;南华大学船山学院, 衡阳, 421001;[戴佩; 邓湘赢; 余敏君; 李玲玲; 罗丹; 曾焱华] 南华大学病原生物学研究所, 特殊病原体防控湖南省重点实验室;[龙娴] 南华大学船山学院, 衡阳, 421001

关键词： T7噬菌体;生殖支原体;互作蛋白

摘要： 目的:从人尿道上皮细胞( SV-HUC-1) T7噬菌体展示cDNA文库筛选生殖支原体黏附蛋白( MgPa)的互作蛋白.方法:以原核表达、纯化的重组生殖支原体黏附蛋白( rMgPa)为靶分子,对人尿道上皮细胞T7噬菌体展示cDNA文库进行4轮生物淘选,采用T7特异性引物扩增阳性克隆的插入片段,PCR产物测序后进行BLAST分析,间接ELISA、斑点免疫印迹和Far-western blot检测阳性噬菌体与rMgPa的特异结合.结果:经过4轮生物淘选,噬菌体克隆富集明显; DNA测序后经BLAST比对分析,结果表明随机挑取的32个阳性克隆包括7种不同序列,其中以RPL35重复次数最多;间接ELISA、斑点免疫印迹和Far-western blot结果表明7个代表性噬菌体均能与rMgPa特异结合.结论:60S核糖体蛋白L35( RPL35)可能是MgPa的互作蛋白,为深入了解MgPa的生物学功能以及生殖支原体的致病机制奠定了实验基础.

语种： 中文

作者： 李玲玲;余敏君;邓湘赢;戴佩;朱翠明;...

期刊： 中南医学科学杂志,2018年46(5):478-481 ISSN：2095-1116

作者机构： 南华大学病原生物学研究所,特殊病原体防控湖南省重点实验室,湖南省子靶标新药研究协同创新中心,湖南 衡阳421001;[廖雅婷; 罗丹; 朱翠明; 戴佩; 余敏君; 李玲玲; 曾焱华; 邓湘赢] 南华大学

关键词： 膜蛋白酵母双杂交;人尿道上皮细胞;cDNA文库

摘要： 利用基于分离的泛素介导的膜蛋白酵母双杂交技术,构建人永生尿道上皮细胞（SV-HUC-1） cDNA文库。Trizol法提取人尿道上皮细胞总RNA,并分离纯化其mRNA;以mRNA为模板,合成cDNA第一链、第二链;通过T4 DNA聚合酶将DNA链两端加上5’接头;并将其大于1 kbp的片段与膜蛋白酵母双杂交载体p PR3-N连接,经电转化大肠杆菌感受态细胞,以构建基于分离的泛素介导的膜蛋白酵母双杂交cDNA文库,并检测文库的库容量和随机性。本研究成功构建了人尿道上皮细胞的基于分离的泛素介导的膜蛋白酵母双杂交cDNA文库,为进一步研究泌尿生殖道感染病原体与宿主尿道上皮细胞的相互作用奠定了实验基础。

语种： 中文

作者： Deng, Xiangying;Dai, Pei;Yu, Minjun;Chen, Liesong;Zhu, Cuiming;...

期刊： International Journal of Medical Microbiology,2018年308(3):405-412 ISSN：1438-4221

通讯作者： Zeng, Yanhua

作者机构： [Zeng, Yanhua] Univ South China, Inst Pathogen Biol, Coll Med, Hengyang 421001, Peoples R China.;Hunan Prov Key Lab Special Pathogens Prevent & Co, Hengyang 421001, Peoples R China.;Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.

通讯机构： [Zeng, Yanhua] U;Univ South China, Inst Pathogen Biol, Coll Med, Hengyang 421001, Peoples R China.

关键词： cyclophilin A;Mycoplasma genitalium adhesion protein;outer membrane protein;recombinant protein;unclassified drug;adhesin;bacterial protein;cyclophilin A;Article;binding affinity;binding assay;controlled study;enzyme linked immunosorbent assay;Far Western blotting;human;human cell;immunofluorescence test;liquid chromatography-mass spectrometry;molecular weight;Mycoplasma genitalium;nonhuman;protein binding;protein localization;protein protein interaction;SV-HUC-1 cell line;Western blotting;bacterium adherence;chemistry;genetics;metabolism;microbiology;Mycoplasma genitalium;mycoplasmosis;pathogenicity;physiology;Adhesins, Bacterial;Bacterial Adhesion;Bacterial Proteins;Blotting, Western;Cyclophilin A;Enzyme-Linked Immunosorbent Assay;Humans;Mycoplasma genitalium;Mycoplasma Infections;Recombinant Proteins

摘要： The Mycoplasma genitalium adhesion protein (MgPa), the most important outer membrane protein of M. genitalium, plays a vital role in the adhesion to and invasion of host cells by M. genitalium. Identification of MgPa receptors will help elucidate the pathogenic mechanism of M. genitalium. However, the receptor protein of MgPa has not been reported to date. In this study, an MgPa-binding protein with a molecular weight of approximately 17 kDa was screened from SV-HUC-1 cell membrane proteins by a modified virus overlay protein binding assay (VOPBA). Liquid chromatography-mass spectrometry (LC–MS) was used to analyze the protein components of the 17-kDa protein. The results demonstrated that the MgPa-binding protein was most likely Cyclophilin A (CyPA). The binding activity and distribution of CyPA in SV-HUC-1 cells were detected using indirect ELISA, western blotting, far-western blotting and indirect immunofluorescence. We found that recombinant MgPa (rMgPa) could bind with CyPA from SV-HUC-1 cell membrane proteins and to recombinant CyPA, which indicated that CyPA was predominant component of the 17-kDa protein band and can interact with rMgPa. In addition, an indirect immunofluorescence assay showed that CyPA was partially distributed on the membrane surfaces of SV-HUC-1 cells and could partially inhibit the adhesion of rMgPa and M. genitalium to SV-HUC-1 cells. Co-localization assays further indicated that rMgPa and M. genitalium can interact with CyPA. These results suggested that the CyPA located on SV-HUC-1 cell membranes may be the potential receptor of MgPa, which could provide an experimental basis for elucidating the function of MgPa and the possible pathogenic mechanism of M. genitalium. © 2018 Elsevier GmbH

语种： 英文

作者： Chen, Zhixi;Chen, Lili;Wang, Chuan;Yu, Jian;Bai, Qinqin;...

期刊： MOLECULAR MEDICINE REPORTS,2017年16(4):4835-4842 ISSN：1791-2997

通讯作者： Wu, Yimou

作者机构： [Yu, Jian; Wu, Yimou; Bai, Qinqin; Hu, Yanqun; Chen, Lili; Song, Yin; Yu, Minjun; Wang, Chuan; Chen, Zhixi] Univ South China, Med Coll, Inst Pathogen Biol, Hunan Prov Key Lab Special Pathogens Prevent & Co, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Chen, Zhixi] Univ South China, Affiliated Hosp 1, Blood Transfus Dept, Hengyang 421001, Hunan, Peoples R China.;[Wu, Yimou; Chen, Lili; Wang, Chuan; Chen, Zhixi] Univ South China, Hunan Cooperat Innovat Ctr Mol Target New Drug St, Hengyang 421001, Hunan, Peoples R China.

通讯机构： [Wu, Yimou] U;Univ South China, Med Coll, Inst Pathogen Biol, Hunan Prov Key Lab Special Pathogens Prevent & Co, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

关键词： cystathionine gamma lyase;gamma interferon;outer membrane protein;periplasmic protein;RNA 16S;tryptophan;gamma interferon;Article;bacterium culture;cell inclusion;controlled study;down regulation;gene expression;gene targeting;genetic transcription;HeLa cell line;host pathogen interaction;nonhuman;ornithosis;persistent infection;reverse transcription polymerase chain reaction;supplementation;transmission electron microscopy;upregulation;cell culture;Chlamydia psittaci;drug effect;fluorescent antibody technique;gene expression regulation;genetics;host pathogen interaction;human;microbiology;ornithosis;ultrastructure;Cells, Cultured;Chlamydophila psittaci;Fluorescent Antibody Technique;Gene Expression Regulation, Bacterial;HeLa Cells;Host-Pathogen Interactions;Humans;Interferon-gamma;Psittacosis;Transcription, Genetic

摘要： The obligate intracellular bacterium Chlamydia psittaci is the causative agent of psittacosis in birds and humans. The capability of this zoonotic pathogen to develop a persistent phase may serve a role in the chronicity of infections, in addition to the failure of antibiotic therapy or immunoprophylaxis. In the present study, a C. psittaci strain 6BC persistent infection cell model was induced using interferon (IFN)-γ, alterations in the infectivity and morphology of the pathogen were analyzed, and the transcript profile of seven selected genes was analyzed. Following treatment with IFN-γ, the infectivity of C. psittaci 6BC was decreased, the inclusion bodies appeared to be smaller, reticulate bodies were larger and the number of infectious elementary bodies was decreased compared with acute infection. In IFN-γ-induced persistently infected cells, the relative mRNA expression levels of the genes CPSIT-0208, CPSIT-0310, CPSIT-0846, CPSIT-0844 and CPSIT-0594 were upregulated at 2-48 h post-infection (p.i.). The genes CPSIT-0959 and CPSIT-0057 were downregulated at 2-36 h p.i. The results of the present study advanced the understanding of C. psittaci persistent infection and demonstrated a number of previously unknown alterations in chlamydial gene expression, which may provide novel targets to further analyze this particular host-pathogen interaction.

语种： 英文

作者： Hu, Jihong;Chen, Chunyan;Ou, Guangli;You, Xiaoxing;Tan, Tianping;...

期刊： Pathogens and Disease,2017年75(4) ISSN：0928-8244

通讯作者： Zhu, Cuiming

作者机构： [Tan, Tianping; Chen, Chunyan; Ou, Guangli; Hu, Jihong; Zhu, Cuiming; Zeng, Yihua; Hu, Xinnian; You, Xiaoxing; Yu, Minjun] Univ South China, Inst Pathogen Biol, Med Coll, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Tan, Tianping; Chen, Chunyan; Ou, Guangli; Hu, Jihong; Zhu, Cuiming; Zeng, Yihua; Hu, Xinnian; You, Xiaoxing; Yu, Minjun] Hunan Prov Key Lab Special Pathogens Prevent & Co, Hengyang 421001, Peoples R China.;[Hu, Jihong; Zhu, Cuiming] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.

通讯机构： [Zhu, Cuiming] U;Univ South China, Inst Pathogen Biol, Med Coll, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

关键词： heme oxygenase 1;immunologic factor;lipid linked protein;NFE2L2 protein, human;nitric oxide;reactive oxygen metabolite;transcription factor Nrf2;biosynthesis;human;immunological tolerance;immunology;inflammation;metabolism;monocyte;Mycoplasma pneumoniae;THP-1 cell line;Heme Oxygenase-1;Humans;Immune Tolerance;Immunologic Factors;Inflammation;Lipid-Linked Proteins;Monocytes;Mycoplasma pneumoniae;NF-E2-Related Factor 2;Nitric Oxide;Reactive Oxygen Species;THP-1 Cells

摘要： A series of inflammatory responses caused by Mycoplasma pneumoniae largely depend on the lipid-associated membrane proteins (LAMPs). Nuclear factor E2-related factor 2 (Nrf2), a transcription factor, is considered to be a critical modulator of inflammatory responses and cellular redox homeostasis. Monocytes play an important role in the invasion and immunity to resist pathogens. Here, we investigated the role of Nrf2 in the anti-inflammatory response stimulated by LAMPs using the human monocyte cell line THP-1. LAMPs were shown to affect the localization of Nrf2, and the levels of reactive oxygen species and inflammatory reactants, including nitric oxide (NO), prostaglandin E2 (PGE2) and cytokines (IL-6, IL-8), were highly elevated in LAMP-stimulated Nrf2-silenced THP-1 cells. Moreover, LAMPs induced the levels of mRNA and the expression of heme oxygenase-1 (HO-1). In summary, our results demonstrated that LAMPs cause nuclear translocation of Nrf2, which further suppresses the expression of inflammatory reactants in THP-1 cells. © Crown copyright 2017.

语种： 英文

作者： Chen, Guodong;Tang, Na;Wang, Chao;Xiao, Linqiao;Yu, Minjun;...

期刊： Biochemical and Biophysical Research Communications,2017年484(2):311-317 ISSN：0006-291X

通讯作者： Zhang, Yan

作者机构： [Han, Liang; Wang, Chao; Chen, Guodong; Tang, Na; Zhao, Lanhua; Cai, Hengling; Xie, Chengyuan; Yu, Minjun; Zhang, Yan; Xiao, Linqiao] Univ South China, Coll Med, Inst Pathogen Biol, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

通讯机构： [Zhang, Yan] U;Univ South China, Coll Med, Inst Pathogen Biol, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

关键词： Helicobacter pylori;TNF-alpha inducing protein (Tip alpha);IL-6/STAT3 pathway;Gastric cancer;Epithelial-mesenchymal transition (EMT)

摘要： Tumor necrosis factor (TNF)-alpha-inducing protein (Tip alpha) is a newly identified carcinogenic factor secreted by Helicobacter pylori (H.pylori). Although it has been proved that Tipa is a strong inducer of epithelialmesenchymal transition (EMT), a crucial process of migration, the exact molecular mechanism is unknown. Current evidence indicates that the oncogenic transcription factor signal transducers and activators of transcription 3 (STAT3) is inappropriately activated in multiple malignancies, including gastric cancer. In this study, we showed that Tipa significantly down-regulated the expression of EMT-related markers E-cadherin as well as up-regulated N-cadherin and vimentin in SGC7901 cells, with typical morphological changes of EMT. Tipa also promoted proliferation and migration of SGC7901 cells. Furthermore, Tipa activated interleukin-6 (IL-6)/STAT3 signaling pathway in SGC7901 cells. The effects of Tipa treatment observed was abolished when we block IL-6/STAT3 signaling pathway. Altogether, our data demonstrated that Tipa may accelerate tumor aggressiveness in gastric cancer by promoting EMT through activation of IL-6/STAT3 pathway. (C) 2017 Elsevier Inc. All rights reserved.

语种： 英文

作者： Ran, Ou;Liang, Mingxing;Yu, Jian;Yu, Minjun;Song, Ying;...

期刊： Pathogens and Disease,2017年75(3) ISSN：0928-8244

通讯作者： Wu Yimou

作者机构： [Wu Yimou; Yu, Minjun; Ran, Ou] Univ South China, Coll Med, Pathogen Biol Inst, Hunan Prov Key Lab Special Pathogens Prevent & Co, Hengyang 421001, Peoples R China.;[Wu Yimou; Ran, Ou] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.;[Ran, Ou] Cent Hosp Xiangtan, Dept Clin Lab, Xiangtan 411100, Peoples R China.;[Liang, Mingxing] First Peoples Hosp Huaihua, Dept Clin Lab, Huaihua 418000, Peoples R China.;[Yu, Jian] Univ South China, Coll Med, Dept Expt Zool, Hengyang 421001, Peoples R China.

通讯机构： [Wu Yimou] U;Univ South China, Coll Med, Pathogen Biol Inst, Hunan Prov Key Lab Special Pathogens Prevent & Co, Hengyang 421001, Peoples R China.

关键词： bacterial DNA;gamma interferon;genomic DNA;immunoglobulin G antibody;protective agent;recombinant protein;recombinant protein cpsit 0846;unclassified drug;bacterial protein;bacterium antibody;cytokine;recombinant protein;animal experiment;animal model;animal tissue;Article;Bagg albino mouse;controlled study;drug efficacy;female;HeLa cell line;human;human cell;humoral immunity;immunization;immunogenicity;in vitro study;in vivo study;lung parenchyma;mouse;nonhuman;ornithosis;priority journal;T lymphocyte;animal;blood;Chlamydia psittaci;disease model;genetics;immunology;lung;metabolism;microbiology;pathology;Psittacosis;Th1 cell;Animals;Antibodies, Bacterial;Bacterial Proteins;Chlamydophila psittaci;Cytokines;Disease Models, Animal;Female;HeLa Cells;Humans;Immunization;Lung;Mice;Mice, Inbred BALB C;Psittacosis;Recombinant Proteins;Th1 Cells

摘要： Chlamydia psittaci is an obligate intracellular bacteria that causes respiratory disease in poultry and humans. Currently, there are no licensed vaccines against chlamydial infection in humans. The transmembrane head protein CPSIT_0846 of C. psittaci is a putative member of the larger Inc protein family. In this study, we investigated immunogenicity and protective efficacy of the recombinant CPSIT_0846 protein in BALB/c mice. Mice immunized with CPSIT_0846 developed strong T-lymphocyte responses that were recalled by the immunogen CPSIT_0846 in an in vitro restimulation assay. These T cells displayed a strong Th1-biased cytokine profile with high levels of IFN-γ. At the same time, a strong humoral immune response was also detected in the immunized mice with high titers of Chlamydia psittaci-specific serum IgG antibodies. More importantly, the robust immune responses correlated well with significantly reduced chlamydial burden and inflammatory pathology in the mouse lungs upon an airway challenge infection. The above results together suggest that the CPSIT_0846 protein may be a potential vaccine candidate antigen for inducing protection against C. psittaci infection and disease in the airway. CPSIT_0846 protein is a potential vaccine candidate against C. psittaci infection for further study. © FEMS 2017. All rights reserved.

语种： 英文

作者： 唐双阳;李乐;李忠玉;曾铁兵;余敏君;...

期刊： 山西医科大学学报,2016年18(11):905-907 ISSN：1007-6611

作者机构： 南华大学医学院病原学实验中心,衡阳,421001;南华大学公共卫生学院放射医学系;[李乐; 唐双阳; 曾铁兵; 李忠玉; 赵兰华; 余敏君; 申海燕] 南华大学

关键词： 医学微生物学;实验教学;教学改革

摘要： 医学微生物学实验课程是验证、巩固和补充课堂理论教学的必要环节,是培养医学生操作技能和创新能力的重要实践环节。文章在充分调研基础上提出了CBL＋TBL双轨制医学微生物实验教学理念,构建了新型实验教学课程体系,对师生角色进行了深入探讨,分层次开放教学科研实验室,完善优化实验教学保障体系,为创新科研型医学人才培养进行了有益的探索。

语种： 中文

作者： Liu, Sheng;Luo, Jingjing;Liu, Yapu;Tang, Shuangyang;Chen, Chaoqun;...

期刊： APMIS,2015年123(7):571-579 ISSN：0903-4641

通讯作者： Zhang, Yan

作者机构： [Liu, Sheng; Chen, Chaoqun; Cai, Hengling; Tang, Shuangyang; Yu, Minjun; Zhang, Yan; Luo, Jingjing; Liu, Yapu] Univ South China, Inst Pathogen Biol, Hengyang 421001, Hunan, Peoples R China.

通讯机构： [Zhang, Yan] U;Univ South China, Inst Pathogen Biol, Hengyang 421001, Hunan, Peoples R China.

关键词： Helicobacter pylori;regulatory T cells;CD4;Foxp3

摘要： <jats:p><jats:italic>Helicobacter pylori</jats:italic> (<jats:italic>H.pylori</jats:italic>), one of the most common infections, is associated with various clinical outcomes. In addition to inducing inflammation, immunological clearance of the pathogen is often incomplete. Regulatory T cells (Treg cells) have been recently demonstrated to play an important role in <jats:italic>H.pylori</jats:italic> infection and the final clinical outcome. The aim of this study was to investigate the number and localization of <jats:styled-content style="fixed-case">CD</jats:styled-content>4<jats:sup>+</jats:sup>Foxp3<jats:sup>+</jats:sup> Treg cells in stomachs and spleens of <jats:italic>H.pylori</jats:italic>‐infected mice. The expression levels of Foxp3 as well as anti‐ and pro‐inflammatory cytokines before and after <jats:italic>H.pylori</jats:italic> triple eradication therapy were examined. We found that the percentages of <jats:styled-content style="fixed-case">CD</jats:styled-content>4<jats:sup>+</jats:sup>Foxp3<jats:sup>+</jats:sup> Treg cells out of the lamina propria lymphocytes (<jats:styled-content style="fixed-case">LPL</jats:styled-content>s) and spleen lymphocytes in the infection group were higher than the <jats:styled-content style="fixed-case">PBS</jats:styled-content> negative control group and the treatment group. <jats:italic>H.pylori</jats:italic> antigen stimulation was associated with an increased number of Treg cells <jats:italic>invitro</jats:italic>. Furthermore, compared with the <jats:styled-content style="fixed-case">PBS</jats:styled-content> and treatment groups, a higher <jats:styled-content style="fixed-case">mRNA</jats:styled-content> expression level of Foxp3 in the gastric tissue was detected in the infection group. <jats:styled-content style="fixed-case">IL</jats:styled-content>‐10 and <jats:styled-content style="fixed-case">TGF</jats:styled-content>‐β1 contents were increased significantly in the culture supernatant of spleen lymphocyte stimulated with <jats:italic>H.pylori</jats:italic> antigen. A marked elevation in serum <jats:styled-content style="fixed-case">IFN</jats:styled-content>‐γ level was observed in <jats:italic>H.pylori</jats:italic>‐infected mice. In addition, gastric tissues of the infection group contained more Foxp3<jats:sup>+</jats:sup> cells. These results indicate that the percentage of <jats:styled-content style="fixed-case">CD</jats:styled-content>4<jats:sup>+</jats:sup>Foxp3<jats:sup>+</jats:sup> Treg cells are increased in <jats:italic>H.pylori‐</jats:italic>infected mice, suggesting a role of Treg cells in <jats:italic>H.pylori‐</jats:italic>induced pathologies, even at the early stages of chronic gastritis and gastric tumorigenesis.</jats:p>

语种： 英文

作者： 唐双阳;李乐;李忠玉;余敏君;刘良专;...

期刊： 当代医学,2015年(22):163-164 ISSN：1009-4393

作者机构： 湖南 421001 南华大学船山学院医学系;南华大学医学院;南华大学公共卫生学院;[李乐] 南华大学船山学院;[唐双阳; 李忠玉; 王可耕; 刘良专; 余敏君] 南华大学

关键词： 独立学院;医学生;示范实验室;实践;创新

摘要： 独立学院医学人才培养模式应注重学生实践技能与创新能力的系统培养,这也是我国医学高等教育改革的重要发展趋势。通过分析我国独立学院医学生人才培养现状,进一步拓展省级示范实验室教学职能,分层次开放实验室,优化实验教学内容,改革传统实验教学方法,调动学生自主学习积极性,切实提升独立学院医学生实践创新能力。

语种： 中文

作者： 何璐;游晓星;李国华;曾焱华;李冉辉;...

期刊： 中南医学科学杂志,2015年(4):364-368 ISSN：2095-1116

作者机构： 南华大学附属第一医院妇产科，湖南 衡阳421001;南华大学医学院病原生物学研究所，特殊病原体防控湖南省重点实验室;南华大学心血管疾病研究所;南华大学附属第一医院妇产科,湖南 衡阳,421001;[李冉辉; 李国华; 吴移谋; 余敏君; 朱翠明; 游晓星; 曾焱华] 南华大学

关键词： 生殖支原体;脂质相关膜蛋白;血红素氧合酶-1;滋养层细胞

摘要： 目的：探讨生殖支原体脂质相关膜蛋白(LAMPs)诱导胎盘滋养层细胞表达血红素氧合酶-1(HO-1)的分子机制。方法用0.5~5μg/mL生殖支原体LAMPs处理体外培养的胎盘滋养层细胞4~12 h,采用实时定量PCR和Western blot法分别检测HO-1 mRNA和蛋白的表达以及核因子相关因子-2(Nrf2)的核转位；比色法观察HO-1的酶活性；2ˊ,7ˊ-二氯二氢荧光黄二乙酸酯( H2 DCFDA)检测活性氧产生。分别采用酪氨酸激酶c-Src抑制剂PP1、活性氧抑制剂N-乙酰半胱氨酸( NAC)和Nrf2 siRNA干预,观察HO-1的表达情况。结果生殖支原体LAMPs能诱导滋养层细胞HO-1 mRNA和蛋白的表达,上调其酶活性。同时,LAMPs也能诱导其产生活性氧,并促使Nrf2核转位。 PP1和NAC预处理后,可明显降低HO-1的表达水平以及细胞核内Nrf2含量。采用Nrf2 siRNA转染后,HO-1的表达显著减少。结论生殖支原体LAMPs通过c-Src/ROS/Nrf2途径诱导滋养层细胞表达HO-1。

语种： 中文

作者： 谭田平;欧广利;刘艳;何军;游晓星;...

期刊： 中华微生物学和免疫学杂志,2015年35(2):112-116 ISSN：0254-5101

通讯作者： Zhu, C.

作者机构： [谭田平; 欧广利; 游晓星; 曾焱华; 余敏君; 朱翠明] 南华大学病原生物学研究所;[何军] 南华大学附属南华医院检验科;[刘艳] 邵阳医学高等专科学校

通讯机构： [Zhu, C.] I;Institute of Pathogen Biology, China

关键词： 肺炎支原体;脂质相关膜蛋白;血红素氧合酶-1

摘要： 目的：研究肺炎支原体（ Mycoplasma pneumonia， Mp）脂质相关膜蛋白（ lipid-associated membrane proteins ， LAMPs）对人单核细胞系THP-1表达血红素氧合酶-1（heme oxygenase 1，HO-1）的影响。方法体外培养THP-1细胞，根据实验目的加入不同浓度LAMPs作用不同时间，并设阳性和阴性对照。收集各处理组细胞，LDH漏出实验检测LAMPs对THP-1细胞的毒性，实时荧光反转录PCR检测 HO-1 mRNA 表达， Western blot 检测 HO-1蛋白表达，比色法检测 HO-1酶活性。结果LAMPs浓度为10μg／ml时，LDH漏出率明显增高。 LAMPs刺激THP-1细胞9 h后HO-1 mRNA表达水平最高；HO-1蛋白表达水平与LAMPs刺激呈剂量依赖性和时间依赖性，其中5．0μg／ml LAMPs刺激表达的HO-1蛋白水平最高；LAMPs刺激12 h 后的HO-1蛋白浓度最高。 LAMPs刺激THP-1细胞后，随着HO-1表达水平的增高，其酶活性亦显著增强。结论 Mp LAMPs可诱导THP-1细胞表达HO-1，并可上调其酶活性。

语种： 中文

作者： Li, Xiang;Liu, Sheng;Luo, Jingjing;Liu, Anyuan;Tang, Shuangyang;...

期刊： Pathogens and Disease,2015年73(4) ISSN：0928-8244

通讯作者： Zhang, Yan

作者机构： [Liu, Sheng; Li, Xiang; Liu, Anyuan; Tang, Shuangyang; Yu, Minjun; Zhang, Yan; Luo, Jingjing; Liu, Shuo] Univ South China, Inst Pathogen Biol, Hengyang 421001, Hunan, Peoples R China.;[Zhang, Yan] Univ South China, Inst Pathogen Biol, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

通讯机构： [Zhang, Yan] U;Univ South China, Inst Pathogen Biol, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.

关键词： carrier protein;inflammasome;interleukin 18;interleukin 18 protein, human;interleukin 18 protein, human;interleukin 1beta;interleukin 1beta converting enzyme;NLRP3 protein, human;NLRP3 protein, human;reactive oxygen metabolite;cell line;enzyme linked immunosorbent assay;flow cytometry;gene expression profiling;Helicobacter pylori;human;immunology;metabolism;microbiology;monocyte;real time polymerase chain reaction;secretion (process);signal transduction;Western blotting;Blotting, Western;Blotting, Western;Carrier Proteins;Carrier Proteins;Caspase 1;Caspase 1;Cell Line;Cell Line;Enzyme-Linked Immunosorbent Assay;Enzyme-Linked Immunosorbent Assay;Flow Cytometry;Flow Cytometry;Gene Expression Profiling;Gene Expression Profiling;Helicobacter pylori;Helicobacter pylori;Humans;Humans;Inflammasomes;Inflammasomes;Interleukin-18;Interleukin-18;Interleukin-1beta;Interleukin-1beta;Monocytes;Monocytes;Reactive Oxygen Species;Reactive Oxygen Species;Real-Time Polymerase Chain Reaction;Real-Time Polymerase Chain Reaction;Signal Transduction;Signal Transduction

摘要： This study investigated whether Helicobacter pylori could activate the nucleotide-binding oligomerization domain-like receptor (NLR) family, pyrin domain-containing 3 (NLRP3) inflammasome in human macrophages and the involvement of reactive oxygen species (ROS) in inflammasome activation. Phorbol-12-myristate-13-acetate (PMA)-differentiated human acute monocytic leukemia cell line THP-1 was infected with H. pylori. The levels of pro-inflammatory cytokines interleukin (IL)-1β and IL-18 in supernatant were measured by ELISA. Intracellular ROS level was analyzed by flow cytometry. Quantitative real-time PCR and western blot analysis were employed to determine the mRNA and protein expression levels of NLRP3 and caspase-1 in THP-1 cells, respectively. Our results showed that H. pylori infection could induce IL-1β and IL-18 production in PMA-differentiated THP-1 cells in a dose- and time-dependent manner. Moreover, secretion of IL-1β and IL-18 in THP-1 cells following H. pylori infection was remarkably reduced by NLRP3-specific small interfering RNA treatment. In addition, the intracellular ROS level was elevated by H. pylori infection, which could be eliminated by the ROS scavenger N-acetylcysteine (NAC). Furthermore, NAC treatment could inhibit NLRP3 inflammasome formation and caspase-1 activation and suppress the release of IL-1β and IL-18 from H. pylori-infected THP-1 cells. These findings provide novel insights into the innate immune response against H. pylori infection, which could potentially be used for the prevention and treatment of H. pylori-related diseases. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

语种： 英文

作者： 何璐;游晓星;李国华;曾焱华;李冉辉;...

期刊： 细胞与分子免疫学杂志,2015年31(2):194-198 ISSN：1007-8738

作者机构： [何璐] Departments of Gynecology and Obstetrics, First Affiliated Hospital, Institute of Pathogenic Biology, Medical College, Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Hengyang 421001, China;[吴移谋; 朱翠明; 游晓星; 余敏君; Zeng, Yanhua; 李冉辉] Institute of Pathogenic Biology, Medical College, Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, Hengyang 421001, China;[李国华] Institute of Cardiovascular Disease, University of South China, Hengyang 421001, China

关键词： 生殖支原体;脂质相关膜蛋白;血红素氧合酶-1;滋养层细胞

摘要： 目的观察生殖支原体脂质相关膜蛋白(LAMP)能否诱导胎盘滋养层细胞表达血红素氧合酶1(HO-1), 从而影响细胞因子的产生。方法体外培养胎盘滋养层细胞, 用0.5~5 μg/mL LAMP作用4~12 h。实时定量PCR和Western blot法分别检测HO-1 mRNA和蛋白的表达以及核因子相关因子2(Nrf2)的核转位; 2′, 7′-二氯二氢荧光黄二乙酸酯(H_2 DCFDA)检测活性氧(ROS)产生; 采用N-乙酰半胱氨酸(NAC)或Nrf2 siRNA处理滋养层细胞, 观察其对HO-1表达的影响。采用HO-1的激动剂钴原卟啉(CoPP), 抑制剂锌原卟啉(ZnPP)或HO-1 siRNA处理细胞, ELISA检测处理前后LAMP对诱导肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)分泌的影响。结果LAMP能诱导滋养层细胞HO-1 mRNA和蛋白表达, 并能诱导其产生ROS以及促进Nrf2核转位。ROS抑制剂NAC预处理后, 可明显降低HO-1的表达水平以及细胞核内Nrf2含量。同时, Nrf2 siRNA转染后, HO-1表达显著减少。采用ZnPP处理滋养层细胞, 或RNA干扰HO-1表达后, 可促进LAMP诱导滋养层细胞分泌TNF-α和IL-1β, 而CoPP处理能进一步降低TNF-α和IL-1β水平。结论LAMP能通过ROS/Nrf2诱导滋养层细胞表达HO-1, 从而抑制细胞因子的过度分泌。

语种： 中文

作者： 谢婧;游晓星;曾焱华;李冉辉;陈列松;...

期刊： 细胞与分子免疫学杂志,2014年30(12):1266-1270 ISSN：1007-8738

作者机构： [谢婧] Departments of Obstetrics, Center Hospital of Hengyang, Hengyang 421001, China;[吴移谋; 陈列松; 朱翠明; 游晓星; 余敏君; Zeng, Yanhua; 李冉辉] Institution of Pathogenic Biology, Medical College, Hengyang 421001, China;[何军] Institution of Pathogenic Biology, Medical College, Department of Clinical Laboratory, Affiliated Nanhua Hospital, University of South China, Hengyang 421001, China

关键词： 生殖支原体;脂质相关膜蛋白;β-防御素2;宫颈上皮细胞

摘要： 目的观察生殖支原体脂质相关膜蛋白（LAMP）能否诱导宫颈上皮细胞表达人β防御素2（hBD-2）,并探讨其可能的调控机制。方法体外培养End1/E6E7人宫颈上皮细胞,用不同浓度的LAMP作用细胞48 h,反转录PCR检测hBD-2 mRNA的表达,Western blot法检测hBD-2蛋白的表达;用Toll样受体2（TLR2）和TLR6中和抗体孵育End1/E6E7细胞,并用TLR2、TLR6和MyD88负显性突变体转染细胞,以明确TLR2、TLR6和MyD88在介导hBD-2表达中的作用;Western blot法检测核因子κB（NF-κB）p65亚基核转位情况,电泳迁移率实验（EMSA）分析LAMP处理前后NF-κB的DNA结合活性;采用NF-κB抑制剂二硫代氨基甲酸吡咯烷（PDTC）处理,观察对hBD-2表达的影响。结果生殖支原体LAMP可诱导End1/E6E7细胞表达hBD-2mRNA和蛋白。TLR2和TLR6中和抗体以及其负显性突变体转染后,能降低hBD-2表达;MyD88负显性突变体也能显著抑制LAMP诱导的hBD-2表达。Western blot结果显示,LAMP处理后可诱导p65核转位,并能增强NF-κB的DNA结合活性。而NF-κB抑制剂PDTC处理后,hBD-2表达降低。结论生殖支原体LAMP可诱导End1/E6E7细胞表达hBD-2,可能受TLR2、TLR6/MyD88/NF-κB调控。

语种： 中文

作者： You, Xiaoxing;Liu, Liangzhuan;Zeng, Yanhua;Li, Ranhui;He, Jun;...

期刊： PLOS ONE,2014年9(7):e103433 ISSN：1932-6203

通讯作者： Wu, Yimou

作者机构： [Jiang, Chuanhao; Wu, Yimou; Liu, Liangzhuan; He, Jun; Ou, Guangli; Zhu, Cuiming; Chen, Liesong; You, Xiaoxing; Yu, Minjun; Zeng, Yanhua; Li, Ranhui; Ma, Xiaohua] Univ South China, Coll Med, Inst Pathogen Biol, Hengyang City, Peoples R China.;[Ma, Xiaohua] Changsha Cent Hosp, Dept Clin Lab, Chagnsha City, Peoples R China.;[He, Jun] Univ South China, Affiliated Nanhua Hosp, Dept Clin Lab, Hengyang City, Peoples R China.

通讯机构： [Wu, Yimou] U;Univ South China, Coll Med, Inst Pathogen Biol, Hengyang City, Peoples R China.

关键词： Transfection;Small interfering RNA;Immune receptor signaling;Inflammation;Phosphorylation;Mycoplasma;Inflammatory diseases;Luciferase

摘要： AIMS: This study is to investigate the mechanisms by which macrophage-activating lipopeptide-2 (MALP-2) induces heme oxygenase (HO)-1, a cytoprotective enzyme that catalyzes the degradation of heme, in human monocytes. METHODS: Human monocytic THP-1 cells were cultured for transient transfection with plasmids and stimulation with MALP-2 for indicative time intervals. After incubation with MALP-2, cells were collected and disrupted, before being tested for promoter activity using luciferase assay. For analysis of proteins, immunoreactive bands were detected using an enhanced chemiluminescence Western blotting system, and the band intensity was measured by densitometryic analysis. For the detection of co-immunoprecipitation, SDS-PAGE was performed and the membranes were probed using respective antibodies. To investigate the cellular localization of NF-E2-related factor 2 (Nrf2), cells underwent immunofluorescence staining and confocal microscopy, and were analyzed using electrophoretic mobility shift assay. RESULTS: MALP-2-induced HO-1 expression and promoter activity were abrogated by transfection with dominant negative (DN) plasmids of TLR2 and TLR6, or their neutralizing antibodies. However, inhibition of MyD88 or transfection with the DN-MyD88 was insufficient to attenuate HO-1 expression. In contrast, mutation or silencing of MyD88 adapter-like (Mal) by DN-Mal or siRNA almost completely blocked HO-1 induction. Btk, c-Src and PI3K were also involved in MALP-2-induced HO-1 expression, as revealed by specific inhibitors LFM-A13, PP1 and LY294002, or by transfection with siRNA of c-Src. MALP-2-induced activation of PI3K was attenuated by transfection with DN mutant of Mal, and by pretreatment with LFM-A13 or PP1. Furthermore, MALP-2 stimulated the translocation of Nrf2 from the cytosol to the nucleus and Nrf2 binding to the ARE site in the HO-1 promoter, which could also be inhibited by pretreatment with a PI3K inhibitor, LY294002. CONCLUSIONS: These results indicated that MALP-2 required TLR2/6, Btk, Mal and c-Src to activate PI3K, which in turn initiated the activation of Nrf2 for efficient HO-1 induction.

语种： 英文

作者： 游晓星;马小华;刘良专;曾焱华;余敏君;...

期刊： 中华微生物学和免疫学杂志,2014年34(6):453-458 ISSN：0254-5101

通讯作者： Wu, Y.

作者机构： 南华大学病原生物学研究所, 衡阳,421001;421001 衡阳，南华大学病原生物学研究所;长沙市中心医院;[何军] 南华大学附属南华医院;[马小华; 蒋传好; 吴移谋; 刘良专; 朱翠明; 余敏君; 游晓星; 曾焱华] 南华大学

通讯机构： [Wu, Y.] I;Institute of Pathogenic Biology, Medical College, University of South China, Hengyang 421001, China

关键词： 支原体巨噬细胞活化脂肽2;血红素氧合酶-1;单核细胞

摘要： 目的：观察支原体巨噬细胞活化脂肽2（macrophage-activating lipopeptide-2， MALP-2）能否诱导人单核细胞系THP-1表达血红素氧合酶-1（hemeoxygenase，HO-1），并探讨其相应的调控机制，以明确机体抵抗支原体感染所致炎性损伤的自我防御机制。方法体外培养THP-1细胞，用不同浓度的MALP-2作用12 h， Western blot检测HO-1的表达；THP-1细胞经TLR2和TLR6中和抗体孵育，或构建TLR2和TLR6负显性突变体转染细胞，以明确TLR2和TLR6在介导HO-1表达中的作用；Western blot检测Akt磷酸化情况，并采用PI3K抑制剂LY294002处理细胞，以证实PI3K参与HO-1表达。同时，分别采用EMSA和免疫荧光观察核转录因子Nrf2的DNA结合活性和核转位，并采用Nrf2 siRNA干扰其表达后，Western blot检测HO-1表达。最后，采用siRNA干扰HO-1表达，或采用HO-1的激动剂钴原卟啉（ cobalt protoporphyrin ，CoPP）处理THP-1细胞，ELISA检测细胞因子TNF-α和IL-1β的分泌。结果 MALP-2能诱导THP-1细胞表达HO-1。且TLR2和TLR6中和抗体以及其负显性突变体转染后，HO-1表达水平显著降低；此外，MALP-2能激活PI3K，PI3K抑制剂能抑制HO-1表达；MALP-2能增强Nrf2的DNA结合活性及核转位，PI3K抑制剂处理后，Nrf2的DNA结合活性以及核转位水平进一步降低。 RNA干扰Nrf2后，HO-1表达显著降低。沉默HO-1表达后，TNF-α和IL-1β分泌增高，而CoPP处理能进一步降低TNF-α和IL-1β水平。结论 MALP-2能诱导THP-1细胞表达HO-1，其机制可能受TLR2、6/PI3K/Nrf2通路调控。 HO-1的表达可在一定程度上负调控细胞因子过度分泌。

语种： 中文