Bai Qian, Cao Jing, Dong Tieli, Tao Feng
Department of Anesthesiology, The Second Affiliated Hospital of Zhengzhou University, Henan, People's Republic of China.
Department of Anatomy, School of Basic Medical Sciences, Zhengzhou University, Henan, People's Republic of China.
J Pain Res. 2020 Oct 28;13:2709-2720. doi: 10.2147/JPR.S278474. eCollection 2020.
Rheumatoid arthritis (RA) leads to pain through alteration of gene expression. Although gene expression alteration in knee cartilage or peripheral blood from RA patients has been identified using microarray, it remains unclear whether long non-coding RNA (lncRNA)-mediated gene regulation occurs in primary sensory neurons of dorsal root ganglia (DRG) during RA-like joint inflammation. In the present study, we aimed to analyze lncRNA and related mRNA profiles in the DRG in a knee joint inflammation rat model.
Complete Freund's adjuvant (CFA) was injected in the rat knee joint for preparing the joint inflammation model. A lncRNA-mRNA microarray of rat DRG was employed for transcriptome analysis. Functional roles of differentially expressed lncRNAs and their related mRNAs in the injured DRG were delineated by bioinformatic analysis.
We observed that expression levels of 9000 lncRNAs were altered on day 7 post-CFA, of which 45.17% were up-regulated and 54.83% were down-regulated. Specifically, 69 lncRNAs (42 up and 27 down) were significantly regulated. We also observed that expression levels of 13,744 mRNAs were altered on day 7 post-CFA, of which 49.67% were up-regulated and 50.33% were down-regulated. Specifically, 102 mRNAs (51 up and 51 down) were significantly regulated. Using quantitative real-time PCR, we verified the changes in differentially expressed lncRNAs in the injured DRG.
These results suggest that microarray-based RNA sequencing can be used to identify altered lncRNAs and relevant mRNAs in the DRG of rats with knee joint inflammation.
类风湿性关节炎(RA)通过基因表达改变导致疼痛。虽然已使用微阵列鉴定了RA患者膝关节软骨或外周血中的基因表达改变,但在类风湿性关节炎样关节炎症期间,背根神经节(DRG)的初级感觉神经元中是否发生长链非编码RNA(lncRNA)介导的基因调控仍不清楚。在本研究中,我们旨在分析膝关节炎症大鼠模型中DRG中的lncRNA和相关mRNA谱。
将完全弗氏佐剂(CFA)注射到大鼠膝关节中以制备关节炎症模型。采用大鼠DRG的lncRNA-mRNA微阵列进行转录组分析。通过生物信息学分析描绘差异表达的lncRNA及其相关mRNA在受损DRG中的功能作用。
我们观察到,在注射CFA后第7天,9000种lncRNA的表达水平发生了改变,其中45.17%上调,54.83%下调。具体而言,69种lncRNA(42种上调,27种下调)受到显著调控。我们还观察到,在注射CFA后第7天,13744种mRNA的表达水平发生了改变,其中49.67%上调,50.33%下调。具体而言,102种mRNA(51种上调,51种下调)受到显著调控。使用定量实时PCR,我们验证了受损DRG中差异表达的lncRNA的变化。
这些结果表明,基于微阵列的RNA测序可用于鉴定膝关节炎症大鼠DRG中改变的lncRNA和相关mRNA。