南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (7): 965-971.doi: 10.12122/j.issn.1673-4254.2021.07.01

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长链非编码RNA Linc00638对类风湿关节炎患者炎症和氧化应激的影响

孙艳秋，刘 健，忻 凌，周 琴，陈晓露，丁 香，张先恒   

  • 出版日期:2021-07-20 发布日期:2021-07-19

The low expression of long non-coding RNA Linc00638 contributes to rheumatoid arthritis progression by regulating inflammation and oxidative stress

  • Online:2021-07-20 Published:2021-07-19

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摘要： 目的 探究类风湿关节炎（RA）患者长链非编码RNA（lncRNA）Linc00638的表达，及其对RA滑膜成纤维细胞（FLS）炎症、氧化应激的影响。方法 收集20例正常人（健康对照组）、35例RA患者（RA组）外周血单个核细胞（PBMCs），RT-qPCR法检测Linc00638表达，并研究其与临床指标的相关性；构建Linc00638过表达质粒和小干扰RNA，转染至RA-FLS中；CCK8检测细胞活力；RT-qPCR法检测Linc00638的过表达和干扰效率。ELISA法检测细胞上清液中白介素-4（IL-4）、白介素-6（IL-6）、活性氧（ROS）、超氧化物歧化 酶（SOD）的表达。结果 与健康对照组相比，RA患者血沉（ESR）、C反应蛋白、类风湿因子（RF）、抗环瓜氨酸肽抗体（Anti-CCP）、IgA、 C4显著升高（P<0.05），Linc00638表达显著降低（P<0.01）；ROC曲线下面积AUC为91.86%，Linc00638的最佳截断值为0.74。Spearman相关性分析表明Linc00638与年龄、病程、DAS28、ESR、C反应蛋白、RF、anti-CCP呈负相关，与IL-4，SOD呈正相关（P<0.05）。关联规则分析表明Linc00638的下降与年龄（>60岁）、病程（>10年）、ESR、RF、anti-CCP的升高，与IL-4、SOD的降低具有强关联。过表达Linc00638能够抑制细胞活力，干扰Linc00638能够提升细胞活力。过表达Linc00638组能够显著升高Linc00638、IL-4、SOD水平（P<0.05），降低IL-6、ROS表达（P<0.05）；si-Linc00638组能显著降低Linc00638、IL-4、SOD表达（P<0.05），升高IL-6、ROS表达（P<0.05）。结论 Linc00638在RA患者中低表达，可能通过调控炎症和氧化应激参与疾病进展。

关键词: 类风湿关节炎；长链非编码RNA；炎症；氧化应激

Abstract: Objective To investigate the expression of long non-coding RNA (lncRNA) Linc00638 in rheumatoid arthritis (RA) and its regulatory role in inflammation and oxidative stress of synovial fibroblasts in RA patients (RA-FLS). Methods Peripheral blood mononuclear cells (PBMCs) were collected from 20 healthy individuals and 35 RA patients for detecting the expression of Linc00638 using RT-qPCR to analyze the correlation of Linc00638 expression with the clinical indicators of RA patients. A Linc00638 overexpression plasmid and siRNA targeting Linc00638 were transfected into RA-FLS, and the changes in cell viability was observed using CCK8 assay; the changes in the expression levels of interleukin-4 (IL-4), IL-6, reactive oxygen species (ROS) and superoxide dismutase (SOD) in the supernatant were detected using ELISA. Results Compared with the healthy control subjects, RA patients had significantly increased ESR, CRP, RF, anti-CCP, IgA, and C4 levels (P<0.05) and significantly decreased Linc00638 expression in the PBMCs (P<0.01). The area under the receiver-operating characteristic (ROC) curve of Linc00638 was 91.86% with the best cut- off value of 0.74 for diagnosis of RA. Spearman correlation analysis showed that Linc00638 expression level was negatively correlated with age, course of disease, DAS28, ESR, CRP, RF and anti-CCP, and positively correlated with IL-4 and SOD levels (P<0.05). Association rule analysis showed that a decreased Linc00638 expression was strongly correlated with an increase of age (>60 years), a longer disease course (>10 years), elevated levels of ESR, RF and anti-CCP, and decreased levels of IL-4 and SOD. In RA-FLS, over-expression of Linc00638 significantly inhibited while Linc00638 interference obviously enhanced the cell viability. Over-expression of Linc00638 also significantly increased the levels of IL-4 and SOD (P<0.05) and decreased the expressions of IL-6 and ROS (P<0.05), while interference of Linc00638 produced the opposite effects in the cells (P<0.05). Conclusion RA patients have low expression levels of Linc00638, which may participate in disease progression by regulating inflammation and oxidative stress.

Key words: rheumatoid arthritis; long non-coding RNA; inflammation; oxidative stress