目的 探讨人巨细胞病毒(human cytomegalovirus,HCMV)编码的hcmv-miR-UL22a-3p在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者血浆及外周血细胞中的水平变化及临床价值。方法 选取2019年10月至2020年1月于东部战区总医院确诊的SLE患者(SLE组)及同期年龄、性别匹配体检健康者(对照组)各49例,收集SLE组及对照组血浆和外周血细胞标本。根据SLE疾病活动指数将SLE组分为活动期与非活动期；根据是否合并肾炎将SLE组分为单纯SLE组和狼疮性肾炎组。采用实时荧光定量PCR(qRT-PCR)技术检测血浆和外周血细胞标本中hcmv-miR-UL22a-3p表达水平。采用受试者工作特征曲线(ROC曲线)、相关性分析、逻辑回归分析血浆和血细胞hcmv-miR-UL22a-3p测定对于SLE的临床价值。结果 qRT-PCR 结果显示,hcmv-miR-UL22a-3p在SLE组血浆中水平低于对照组 [ 0.00067(0.00026,0.0016) vs 0.0021(0.0015,0.0043),P＜0.001 ] ；在患者血细胞中水平高于对照组 [ 0.0031(0.0016,0.0045) vs 0.0016(0.0011,0.0023),P＜0.001 ] ,且狼疮性肾炎患者血细胞中hcmv-miR-UL22a-3p水平高于单纯SLE组 [ 0.0042(0.0024,0.015) vs 0.0024(0.0014,0.0042),P＜0.05 ] 。相关性分析显示,血浆hcmv-miR-UL22a-3p水平与白细胞数目呈显著正相关(r = 0.309,P＜0.05 )；血细胞hcmv-miR-UL22a-3p水平与患者血沉呈显著正相关,与补体C3、血白蛋白水平、血白细胞数目呈显著负相关(r分别为0.321,-0.352,-0.295和-0.327,P均＜0.05)。血浆hcmv-miR-UL22a-3p水平区分SLE与对照的ROC曲线下面积(AUC)为 0.792(95% CI：0.701 ~ 0.883),敏感性为75.5%,特异性为 79.6%；血细胞hcmv-miR-UL22a-3p水平区分SLE与对照的AUC为0.744(95% CI：0.647 ~ 0.841),敏感性为 69.4%,特异性为 63.3%；血细胞和血浆hcmv-miR-UL22a-3p水平比值联合区分SLE与对照的 AUC 为0.830(95% CI：0.751 ~ 0.910),敏感性为 71.4%,特异性为 85.7%。多因素Logistic回归分析显示,在校正年龄、性别影响后,血浆hcmv-miR-UL22a-3p水平的降低( OR = 0.106,95% CI = 0.033~0.343,P＜0.001)以及血细胞hcmv-miR-UL22a-3p水平的升高( OR = 10.852,95% CI = 1.830~64.350,P＜0.05)与SLE的发生密切相关。结论 SLE患者血浆hcmv-miR-UL22a-3p水平下调,血细胞hcmv-miR-UL22a-3p水平上调,可作为SLE的潜在辅助诊断生物标志物。
Abstract: Objective To investigate the levels and clinical value of human cytomegalovirus ( HCMV ) -encoded miR-UL22a-3p( hemv-miR-UL22a-3p) in plasma and peripheral blood cells of patients with systemic lupus erythematosus ( SLE). Methods The plasma and peripheral blood cell samples were collected from 49 SLE patients diagnosed in Eastern Theater General Hospital of PLA during October 2019 and January 2020, and 49 age and gender-matched healthy controls, respectively. SLE patents were div ided into active stage and non-active stage according to disease activity index of SLE, and into simple SLE and lupus nephritis according to whether nephritis was combined. The expression levels of hemv-miR-UL22a-3p in plasma and peripheral blood cells were detected by real-time fluorescence quantitative polymerase chain reaction ( qRT-PCR). The clinical value of hemv-miR-UL22a-3p in the diagnosisof SLE was evaluated by the receiver operating characteristic ( ROC) curve, correlation analysis and logistic regression analysis. Results The expression levels of hemv-miR-UL22a-3p in plasma of SLE patients were significantly lower than that in healthy controls (0.000 67 [0.000 26, 0.001 6] vs 0.002 1 [0.001 5, 0.004 3], P<0.001), while those in peripheral blood cells were inverse (0.003 1 [0.001 6, 0.0045] vs 0.001 6[0.001 1, 0.002 3], P<0.001). Moreover, the expression levels of hemv-miR -UL22a-3p in peripheral blood cells of patients with lupus nephritis were significantly higher than that in patients with simple SLE (0.004 2 [0.002 4,0.015] vs 0.0024 [0.001 4,0.004 2], P<0.05). The correlation analysis showed that the levels of plasma hemv-miR-UL22a-3p were positively correlated with the number of white blood cells ( WBC) (r=0.309,P<0.05), and that the levels of hemv -miR-UL22a-3p in peripheral blood cells were positively correlated with erythrocyte sedimentation rate ( ESR)，and negatively with serum complement C3 and albumin ( ALB) levels and the number of WBC (r=0.321, -0.352, -0.295 and -0.327, respectively, P<0.05 ). The area under the ROC curve ( AUCHOC )，sensitivity and specifceity of plasma hemv-miR-UL22a-3p in the diagnosis of SLE were 0.792 (95% CI:0.701-0.883), 75.5% and 79.6% , respectively. The AUCROC，sensitivity and specificity of peripheral blood cell hemv-miR-UL22a-3p in the diagnosis of SLE were 0.744 (95%CI: 0.647-0.841), 69.4% and 63.3%, respectively. The AUCO， sensitivity and specificity of the ratio of peripheral blood cell and plasma hemv-miR-UL22a-3p in the diagnosis of SLE were 0.830 (95%CI: 0.751-0.910),71. 4% and 85.7% , respectively. Multivariate logistic regression analysis showed that the decreased plasma hemv-miR-UL22a-3p level ( 0R=0.083, 95%CI: 0.027-0.250, P<0.001), increased peripheral blood cell hemv -miR-UL22a-3p level (OR= 17.537 , 95% CI:3.748-82.047, P<0.001) and inereased ratio of hemv-rmiR-UL.22a-3p in blood cells and plasma (OR= 1.676, 95%CI: 1.223-2.297,P<0.001) were closely related to the occurrence of SLE after adjusting age and gender. Conclusion The levels of plasma hemv -miR-UL22a 3p are down- regulated and the levels of blood cell hemv -miR-UL22a-3p are up-regulated in patients with SLE. The ratio of hemv-miR- UL22a-3p in blood cells and plasma may serve as a potential biomarker for the diagnosis of SLE.