脓毒症并发急性肾损伤患者外周血单个核细胞中微小RNA-16-5p、干扰素诱导跨膜蛋白3表达及临床意义

Expression and clinical significance of miR-16-5p and IFITM3 in peripheral blood mononuclear cells of patient with sepsis-associated acute kidney injury

目的检测脓毒症及脓毒症并发急性肾损伤(sepsis-associated acute kidney injury,S-AKI)患者外周血单个核细胞中微小RNA-16-5p(microRNA-16-5p,miR-16-5p)、干扰素诱导跨膜蛋白3(interferon-induced transmembrane protein,IFITM3)表达情况,分析两者对S-AKI患者的临床意义。方法本研究以2019年2月至2021年2月武汉市蔡甸区人民医院确诊的脓毒症患者为研究对象,共纳入106例,将合并急性肾损伤(acute kidney injury,AKI)的脓毒症患者作为AKI组(52例),未合并AKI的患者为非AKI组(54例)。收集所有患者的血液样本3 mL并分离单核细胞,利用实时荧光定量技术检测外周血单个核细胞中miR-16-5p、IFITM3 mRNA的表达水平,酶联免疫吸附法测定外周血中白细胞介素1β(interleukin-1β,IL-1β)水平,全自动生化分析仪检测外周血中胱抑素(cysteine,Cys)和尿素氮(blood urea nitrogen,BUN)水平;比较两组间的临床指标及miR-16-5p、IFITM3 mRNA表达水平,Pearson法分析miR-16-5p、IFITM3 mRNA分别与BUN、Cys水平的相关性,以及miR-16-5p与IFITM3 mRNA表达水平的相关性,采用多元线性回归分析S-AKI患者miR-16-5p、IFITM3 mRNA表达水平的影响因素,受试者工作特性曲线(receiver operator characteristic curves,ROC)评估miR-16-5p和IFITM3 mRNA对脓毒症患者发生S-AKI的诊断价值。结果AKI组患者急性生理学与慢性健康状况评分系统Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分(15.67±4.39比13.31±3.56)、序贯器官衰竭评分(sequential organ failure assessment,SOFA)评分(5.62±1.28比4.89±1.41)、IL-1β[(22.03±5.78)mg/L比(10.44±3.05)mg/L]、Cys[(2.72±0.45)mg/L比(1.98±0.31)mg/L]和BUN[(7.93±2.24)μmol/L比(4.49±2.07)μmol/L]水平均高于非AKI组患者(P<0.05),AKI组患者的miR-16-5p(1.64±0.30比1.17±0.28)表达水平显著上调,IFITM3 mRNA(2.87±0.62比4.06±0.92)及蛋白(1.63±0.41比3.15±0.85)(r=0.560,P<0.05)水平呈正相关,IFITM3 mRNA表达水平与IL-1β(r=-0.754,P<0.05)、Cys(r=-0.554,P<0.05)、BUN(r=-0.695,P<0.05)水平呈负相关,miR-16-5p与IFITM3 mRNA的表达水平呈负相关(r=-0.496,P<0.05);多元线性回归分析结果显示,IL-1β升高、Cys升高是miR-16-5p、IFITM3 mRNA表达水平的影响因素(P<0.05);miR-16-5p、IFITM3 mRNA单一检测诊断脓毒症患者发生S-AKI的曲线下面积(area under the curve,AUC)分别为0.876(95%CI:0.809~0.944)、0.874(95%CI:0.811~0.938),miR-16-5p和IFITM3 mRNA联合检测的AUC为0.956(95%CI:0.929~0.992)。结论S-AKI患者中miR-16-5p表达上调,IFITM3表达下调,与S-AKI的病理过程密切相关,可能通过促进炎症反应加重肾损伤,有助于早期诊断S-AKI,为临床治疗S-AKI提供了有希望的靶点。

Objective To detect the expression of microRNA-16-5p(miR-16-5p)and interferon-induced transmembrane protein 3(IFITM3)in peripheral blood mononuclear cells of septic patients with sepsis-associated acute kidney injury(S-AKI)and examine the clinical significance of the two in S-AKI patients.Methods During February 2019 to February 2021,106 septic patients were selected as research subjects.Septic patients with acute kidney injury(AKI)were included in AKI group(n=52)and those without AKI in non-AKI group(n=54).The 3 ml of blood samples from all patients was harvested for separating monocytes,real-time fluorescent quantitative technology for detecting the expression levels of miR-16-5p and IFITM3 mRNA in peripheral blood mononuclear cells,enzyme-linked immunosorbent assay for determining the level of interleukin-1β(IL-1β)in peripheral blood,automatic biochemical analyzer for detecting the levels of cysteine(Cys)and blood urea nitrogen(BUN)in peripheral blood;clinical parameters and the expression levels of miR-16-5p and IFITM3 mRNA were compared between two groups,Pearson’s method was performed to analyze the correlation between miR-16-5p,IFITM3 mRNA and BUN/Cys level and the correlation between miR-16-5p and IFITM3 mRNA expression levels,multiple linear regression was utilized for examining the influencing factors of miR-16-5p and IFITM3 mRNA expression,receiver operator characteristic(ROC)curve was plotted for evaluating the diagnostic value of miR-16-5p and IFITM3 mRNA for S-AKI in septic patients.Results The acute physiology and chronic health evaluationⅡ(APACHEⅡ)score(15.67±4.39 vs 13.31±3.56),sequential organ failure assessment(SOFA)score(5.62±1.28 vs 4.89±1.41),levels of IL-1β[(22.03±5.78)mg/L vs(10.44±3.05)mg/L],Cys[(2.72±0.45)mg/L vs(1.98±0.31)mg/L]and BUN[(7.93±2.24)μmol/L vs(4.49±2.07)μmol/L]were higher in AKI group than those in non-AKI group(P<0.05),the expression level of miR-16-5p(1.64±0.30 vs 1.17±0.28)in AKI group was significantly up-regulated,the expression level of IFITM3 mRNA(2.87±0.62 vs 4.06±0.92)and protein(1.63±0.41 vs 3.15±0.85)were significantly down-regulated(P<0.05);the expression level of miR-16-5p in patient with S-AKI was positively correlated with the levels of IL-1β(r=0.531,P<0.05),Cys(r=0.482,P<0.05)and BUN(r=0.560,P<0.05),the expression level of IFITM3 mRNA was negatively correlated with the levels of IL-1β(r=-0.754,P<0.05),Cys(r=-0.554,P<0.05)and BUN(r=-0.695,P<0.05),the expression levels of miR-16-5p and IFITM3 mRNA were negatively correlated(r=-0.496,P<0.05);multiple linear regression analysis indicated that elevations of IL-1βand Cys were the influencing factors of miR-16-5p and IFITM3 mRNA expression levels(P<0.05);the area under the curve(AUC)s of miR-16-5p and IFITM3 mRNA single test for diagnosing S-AKI in septic patients were 0.876(95%CI:0.809~0.944)and 0.874(95%CI:0.811~0.938),the AUC of miR-16-5p and IFITM3 mRNA combined detection was 0.956(95%CI:0.929~0.992).Conclusion In S-AKI patients,the expression of miR-16-5p is up-regulated while the expression of IFITM3 down-regulated.Both are closely correlated with the pathological process of S-AKI.It may aggravate kidney injury by promoting inflammation,aid in an early diagnosis of S-AKI and provide a promising therapeutic target for S-AKI.