初发系统性红斑狼疮患者外周血单个核细胞中circ-0007385表达变化及其意义

The Expression of circ-0007385 in Peripheral Blood Mononuclear Cells of Patients with Primary Systemic Lupus Erythematosus and Its Significance

摘要: 背景：系统性红斑狼疮(Systemic Lupus Erythematosus, SLE)是一种复杂的多器官多系统受累的自身免疫性疾病，其病因尚未阐明。已有研究表明环状RNA (circRNA)在人体组织和器官中广泛表达，作为内源竞争RNA (ceRNA)与微小RNA (miRNA)结合，在SLE中发挥重要作用。课题组前期通过高通量测序分析筛查SLE患者血清circRNA提示circ-0007385表达异常，可能参与SLE发病。目的：本研究旨在探究SLE患者外周血单个核细胞(PBMC)中circ-0007385的表达水平并分析其意义，为SLE的早期诊断寻找新型生物标志物。方法：选取初发SLE患者60例(活动期34例、稳定期26例)作为SLE组，健康人30例作为对照组。通过高通量测序分析受试者外周血PBMC中的circRNA表达水平，采用RT-qPCR法检测所有受试者血清circ-0007385、miR-181b的表达水平，生物信息技术构建circRNA-miRNA相互作用的网络，采用受试者工作特征(Receiver Operating Characteristic, ROC)曲线分析血清circ-0007385、miR18-1b对于初发SLE诊断的特异性、敏感度和曲线下面积(Area Under the Curve, AUC)。结果：1) SLE患者外周血PBMC中circ-0007385较对照组显著升高(p < 0.01)，且与疾病活动度指标dsDNA抗体正相关(r = 0.6839 p < 0.001)和SLE疾病活动指数(SLEDAI-2K)评分正相关(r = 0.5933 p < 0.01)，与补体C3负相关(r = −0.6617 p < 0.05)。2) 生物信息技术预测circ-0007385与miR-181b特异结合位点，RT-qPCR结果显示，SLE患者外周血PBMC中miR-181b的表达量与对照组相比显著降低(p < 0.01)，并与circ-0007385表达量成负相关(r = −0.5839 p < 0.01)。3) ROC曲线分析表明，外周血PBMC中circ-0007385和miR-181b表达水平可以区分初发SLE患者与对照组，二元逻辑回归模型表明，circ-0007385联合miR-181b可以提供更佳的诊断准确性，曲线下面积(AUC)为0.9078 (敏感性为0.85，特异性0.87)。结论：本研究表明，外周血PBMC中circ-0007385联合miR-181b在预测SLE发病中具有一定的诊断价值，并且与初发SLE患者疾病活动度显著相关。

Abstract: Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease involving multiple organs and multiple systems, and its etiology has not been clarified. Previous studies have shown that circle RNA (circRNA) is widely expressed in human tissues and organs, and plays an important role in SLE as a competitive endogenous RNA (ceRNA) binding to microRNAs (miRNA). In the early stage, the research group screened the serum circRNA of SLE patients through high-throughput sequencing analysis, suggesting that circ-0007385 expression was abnormal, which may be involved in the pathogenesis of SLE. Objective: This study aims to explore the circ-0007385 in peripheral blood mononuclear cells (PBMC) of patients with SLE. The expression level of circ-0007385 and its significance were analyzed to find new biomarkers for early diagnosis of SLE. Methods: 60 patients with primary SLE (34 patients in active phase, 26 patients in stable phase) were selected as SLE group, and 30 healthy people as control group. The circRNA expression level in peripheral blood PBMC of the subjects was analyzed by high-throughput sequencing. The expression level of circ-0007385 and miR-181b in serum of all subjects was detected by RTqPCR. The circRNA-miRNA interaction network was constructed by bioinformatics technology. The specificity of circ-0007385 and miR-181b in serum for the diagnosis of primary SLE was analyzed by receiver operating characteristic (ROC) curve results of sensitivity and area under the curve (AUC). Results: 1) Circ-0007385 in peripheral blood PBMC of SLE patients was significantly higher than that of the control group (p < 0.01), and was positively correlated with dsDNA antibody (r = 0.6839 p < 0.001) and SLEDAI-2K score (r = 0.5933 p < 0.01), negative correlation with complement C3 (r = −0.6617 p < 0.05). 2) Bioinformatics predicted the specific binding sites of circ-0007385 and miR-181b. RT-qPCR results showed that the expression of miR-181b in peripheral blood PBMC of SLE patients was significantly lower than that of control group (p < 0.01), and compared with that of circ-0007385 expression was negatively correlated (r = −0.5839 p < 0.01). 3) ROC curve analysis showed that circ in peripheral blood PBMC circ-0007385 and miR-181b expression levels can distinguish patients with primary SLE from the control group. The binary logistic regression model shows that circ-0007385 combined with miR-181b can provide better diagnostic accuracy, and the area under the curve (AUC) is 0.9078 (sensitivity is 0.85, specificity is 0.87). Conclusion: This study shows that circ-0007385 combined with miR-181b in peripheral blood PBMC has certain diagnostic value in predicting the incidence of SLE, and is significantly related to the disease activity of patients with primary SLE.