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

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

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。

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,andpositively 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,overexpression 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.