类风湿关节炎患者外周血单核细胞lncR NA差异表达谱研究
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摘要
目的:观察类风湿关节炎患者外周血单核细胞lncRNA差异表达。方法:选取安徽中医药大学第一附属医院住院类风湿关节炎患者(类风湿关节炎组)及健康体检者(正常对照组),每组3例。抽取2组研究对象的外周血,利用高通量基因测序技术检测外周血单个核细胞lncRNA和mRNA表达,GO分析及KEGG信号通路分析差异表达的lncRNA功能分布,构建lncRNA-mRNA的共表达网络,并在此基础上通过Cis和Trans预测可能与类风湿关节炎凋亡、氧化应激、信号通路相关lnc RNA。利用RT-PCR方法验证lncRNA。结果:通过lncRNA测序分析发现,lncRNA在类风湿关节炎组与正常对照组存在明显差异表达。共有9158条差异表达的lncRNA(差异倍数≥2,且P≤0.05),表达上调的有5682条,下调的有3476条;差异表达的mRNA共有7895条。GO分析发现,差异表达的mRNA主要参与免疫炎症反应、细胞凋亡、氧化应激、细胞代谢、细胞转录调节、骨髓白细胞活化、血小板细胞活化等。KEGG信号通路分析发现,差异表达的mRNA主要参与类风湿关节炎、细胞衰老、信号通路等方面。通过RT-PCR验证发现,与类风湿关节炎相关的细胞凋亡lncRNA为ENST00000619282、ENST00000637683、ENST00000511703,氧化应激为HOTAIRM1、DANCR,Notch通路为MALAT1、CARMN、LINC-PINT。结论:类风湿关节炎患者外周血淋巴细胞中差异表达的lncRNA异常表达,可能与类风湿关节炎细胞凋亡、氧化应激及信号通路失衡有关。
Objective:To observe the differential expression of lncRNA in peripheral blood mononuclear cells of patients with rheumatoid arthritis(RA).Methods:The RA patients(RA group)and the healthy controls(NC group)in the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were selected,3 cases in each group.Peripheral blood of RA group and NC group were used to detect lncRNA and mRNA expression in peripheral blood mononuclear cells by high-throughput gene sequencing technology.GO and KEGG Pathway analyzed the differential expression of lncRNA function,and constructed a coexpression network of lncRNA-mRNA.Based on this,Cis and Trans predicted lncRNAs that may be associated with RA apoptosis,oxidative stress,and signaling pathways.The lncRNA was verified by RT-PCR.Results:lncRNA sequencing analysis revealed that lncRNA was significantly different between RA and NC groups.There were 9158 differentially expressed lncRNAs(differential multiples ≥ 2,and P ≤ 0.05),5682 expressions were up-regulated,3476 were down-regulated,and 7895 differentially expressed mRNAs.GO analysis found that differentially expressed mRNA is mainly involved in immune inflammatory response,apoptosis,oxidative stress,cellular metabolism,cellular transcriptional regulation,bone marrow leukocyte activation,and platelet cell activation.KEGG pathway analysis found that differentially expressed mRNA is mainly involved in rheumatoid arthritis,cellular senescence,signaling pathways and the like.RT-PCR confirmed that the lncRNAs with RA cells were ENST00000619282,ENST00000637683 and ENST00000511703.The oxidative stress was HOTAIRM1,DANCR.The Notch pathway was MALAT1,CARMN,LINC-PINT.Conclusion:Abnormal expression of lncRNA in peripheral blood lymphocytes of RA patients may be related to RA cell apoptosis,oxidative stress and signal pathway imbalance.
Objective:To observe the differential expression of lncRNA in peripheral blood mononuclear cells of patients with rheumatoid arthritis(RA).Methods:The RA patients(RA group)and the healthy controls(NC group)in the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were selected,3 cases in each group.Peripheral blood of RA group and NC group were used to detect lncRNA and mRNA expression in peripheral blood mononuclear cells by high-throughput gene sequencing technology.GO and KEGG Pathway analyzed the differential expression of lncRNA function,and constructed a coexpression network of lncRNA-mRNA.Based on this,Cis and Trans predicted lncRNAs that may be associated with RA apoptosis,oxidative stress,and signaling pathways.The lncRNA was verified by RT-PCR.Results:lncRNA sequencing analysis revealed that lncRNA was significantly different between RA and NC groups.There were 9158 differentially expressed lncRNAs(differential multiples ≥ 2,and P ≤ 0.05),5682 expressions were up-regulated,3476 were down-regulated,and 7895 differentially expressed mRNAs.GO analysis found that differentially expressed mRNA is mainly involved in immune inflammatory response,apoptosis,oxidative stress,cellular metabolism,cellular transcriptional regulation,bone marrow leukocyte activation,and platelet cell activation.KEGG pathway analysis found that differentially expressed mRNA is mainly involved in rheumatoid arthritis,cellular senescence,signaling pathways and the like.RT-PCR confirmed that the lncRNAs with RA cells were ENST00000619282,ENST00000637683 and ENST00000511703.The oxidative stress was HOTAIRM1,DANCR.The Notch pathway was MALAT1,CARMN,LINC-PINT.Conclusion:Abnormal expression of lncRNA in peripheral blood lymphocytes of RA patients may be related to RA cell apoptosis,oxidative stress and signal pathway imbalance.
引文
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[18]刘健,章平衡,黄旦,等.lncRNA在风湿病中的研究进展[J].风湿病与关节炎,2018,7(12):58-63.
[19]KUNDU S,GHOSH P,DATTA S,et al.Oxidative stress as a potential biomarker for determining disease activity in patients with rheumatoid arthritis[J].Free Radic Res,2012,46(12):1482-1489.
[20]STAVROPOULOS-KALINOGLOU A,DELI C,KITASGD,et al.Muscle wasting in rheumatoid arthritis:The role of oxidative stress[J].World J Rheumatol,2014,4(3):44-53.
[21]ZHANG HJ,WEI QF,WANG SJ,et al.LncRNA HO-TAIR alleviates rheumatoid arthritis by targeting miR-138 and inactivating NF-κB pathway[J].Int Immunopharmacol,2017(50):283-290.
[22]PAN F,ZHU L,LV H,et al.Quercetin promotes the apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by upregulating lncRNA MALAT1[J].Int JMol Med,2016,38(5):1507-1514.
[23]WU J,ZHANG H,ZHENG Y,et al.The Long Noncoding RNA MALAT1 Induces Tolerogenic Dendritic Cells and Regulatory T Cells via miR155/Dendritic Cell-Specific Intercellular Adhesion Molecule-3 Grabbing Nonintegrin/IL10 Axis[J].Front Immunol,2018,13(9):1847.
[2]ZHAO D,JIANG Z,WANG Z,et al.Retinoid interferon-induced mortality19(GRIM9)inhibits proliferation and invasion in rheumatoid arthitis fibroblast-like synoviocytes[J].Biomed Pharmacother,2018(98):719-725.
[3]WAN L,LIU J.Changes of CD4+CD25+Regulatory TCells,FoxP3 in Adjuvant Arthritis Rats with Damage of Pulmonary Function and Effects of Tripterygium Glycosides Tablet[J].Int J Rheumatol,2012(318):348450.
[4]曾小峰,朱松林,谭爱春,等.我国类风湿关节炎疾病负担和生存质量研究的系统评价[J].中国循证医学杂志,2013,13(3):300-307.
[5]夏燕,冯佳,陈安平,等.类风湿关节炎外周血lncRNA差异表达研究[J].中国免疫学杂志,2016,32(1):9-12.
[6]袁敏,徐黎明,郑娟,等.lncRNA在类风湿关节炎患者外周血单个核细胞中的异常表达[J].山东医药,2018,58(10):63-65.
[7]ZHAO Y,LIU Y,LIN L,et al.The lncRNA MACC1-AS1promotes gastric cancer cell metabolic plasticity via AMPK/Lin28 mediated mRNA stability of MACC1[J].Mol Cancer,2018,17(1):69.
[8]LIANG H,PAN Z,ZHAO X,et al.LncRNA PFL contributes to cardiac fibrosis by acting as a competing endogenous RNA of let-7d[J].Theranostics,2018,8(4):1180-1194.
[9]HUANG W,THOMAS B,FLYNN RA,et al.Retraction Note:DDX5 and its associated lncRNA Rmrp modulate TH17 cell effector functions[J].Nature,2018,562(7725):150.
[10]李有强,张云燕,陈茶,等.3-O-C12-HSL阻碍MoDCs成熟过程中lncRNA表达谱的变化[J].重庆医科大学学报,2018,43(1):117-121.
[11]崔桂荣,李焱,贾振薇,等.LncRNA AK093987在弥漫大B细胞淋巴瘤患者中的表达及其临床意义[J].肿瘤学杂志,2018,24(4):360-364.
[12]邓桢,姚芳苡,叶建青,等.长链非编码RNA在脂多糖诱导人巨噬细胞炎症反应中的表达变化[J].中华危重病急救医学,2017,29(4):306-310.
[13]YANG KY,CHEN DL.Shikonin Inhibits Inflammatory Response in Rheumatoid Arthritis Synovial Fibroblasts via lncRNA-NR024118[J].Evid Based Complement Alternat Med,2015,2015:631737.
[14]ZHANG HJ,WEI QF,WANG SJ,et al.LncRNA HO-TAIR alleviates rheumatoid arthritis by targeting miR-138 and inactivating NF-κB pathway[J].Int Immunopharmacol,2017(50):283-290.
[15]LI N,MA T,HAN J,et al.Increased apoptosis induction in CD4+CD25+Foxp3+T cells contributes to enhanced disease activity in patients with rheumatoid arthritis through IL-10 regulation[J].Eur Rev Med Pharmacol Sci,2014,18(1):78-85.
[16]TOUBI E,KESSEL A,MAHMUDOV Z,et al.Increased Spontaneous Apoptosis of CD4+CD25+T Cells in Patients with Active Rheumatoid Arthritis Is Reduced by Infliximab[J].Ann N Y Acad Sci,2010,1051(1):506-514.
[17]曹永贺,刘健.类风湿关节炎患者细胞凋亡与临床指标相关性分析[J].中国中西医结合杂志,2016,36(1):35-39.
[18]刘健,章平衡,黄旦,等.lncRNA在风湿病中的研究进展[J].风湿病与关节炎,2018,7(12):58-63.
[19]KUNDU S,GHOSH P,DATTA S,et al.Oxidative stress as a potential biomarker for determining disease activity in patients with rheumatoid arthritis[J].Free Radic Res,2012,46(12):1482-1489.
[20]STAVROPOULOS-KALINOGLOU A,DELI C,KITASGD,et al.Muscle wasting in rheumatoid arthritis:The role of oxidative stress[J].World J Rheumatol,2014,4(3):44-53.
[21]ZHANG HJ,WEI QF,WANG SJ,et al.LncRNA HO-TAIR alleviates rheumatoid arthritis by targeting miR-138 and inactivating NF-κB pathway[J].Int Immunopharmacol,2017(50):283-290.
[22]PAN F,ZHU L,LV H,et al.Quercetin promotes the apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by upregulating lncRNA MALAT1[J].Int JMol Med,2016,38(5):1507-1514.
[23]WU J,ZHANG H,ZHENG Y,et al.The Long Noncoding RNA MALAT1 Induces Tolerogenic Dendritic Cells and Regulatory T Cells via miR155/Dendritic Cell-Specific Intercellular Adhesion Molecule-3 Grabbing Nonintegrin/IL10 Axis[J].Front Immunol,2018,13(9):1847.