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.
作者
刘健
万磊
孙玥
章平衡
忻凌
姜辉
方妍妍
董文哲
文建庭
LIU Jian;WAN Lei;SUN Yue;ZHANG Ping-heng;XIN Ling;JIANG Hui;FANG Yan-yan;DONG Wen-zhe;WEN Jian-ting
出处
《风湿病与关节炎》
2019年第2期10-15,共6页
Rheumatism and Arthritis
基金
中华人民共和国科技部中医药现代化研究(2018YFC1705200)
安徽高校自然科学研究重点项目(KJ2017A281)
安徽省教育厅高校自然科学研究重点项目(KJ2018A0279)
2016年中央引导地方科技发展专项(财教[2016]1188)
安徽省科技攻关项目(1604a0802085)
现代中医内科应用基础与开发研究安徽省重点实验室(财教[2016]518)
2015年技术创新服务体系专项(财教[2015]1551)
