长链非编码RNA H19在肺炎支原体肺炎病儿血清中的表达及其意义

Expression and significance of long noncoding RNA H19 in the serum of children with mycoplasma pneumoniae pneumonia

目的探讨长链非编码RNA H19(LncRNA H19)在肺炎支原体肺炎(MPP)病儿血清中的表达及其对肺炎支原体脂质相关膜蛋白(LAMPS)诱导的肺泡巨噬细胞凋亡和炎性因子表达的影响。方法收集2017年10月至2018年12月南阳市中心医院收治的MPP病儿60例为观察组,根据病儿病情程度分为恢复期(30例)与急性期(30例);选取同期到我院体检的健康儿童60例作为对照组。实时荧光定量聚合酶链反应(qRT-PCR)检测LncRNA H19的表达水平;采用受试者工作特征(ROC)曲线分析LncRNA H19对MPP的诊断价值。体外培养小鼠肺泡巨噬细胞MH-S,LAMPS处理细胞建立细胞损伤模型。分别将si-NC、si-H19转染至MH-S细胞,使用LAMPS处理24 h。流式细胞术检测细胞凋亡率;ELISA法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素6(IL-6)水平。结果与对照组相比,观察组血清LncRNA H19的表达水平显著升高[(1.01±0.13)比(2.58±0.25),P<0.05];与MPP恢复期相比,MPP急性期血清LncRNA H19的表达水平显著升高[(1.63±0.20)比(3.24±0.39),P<0.05];ROC分析显示LncRNA H19在MPP诊断中敏感度为86.67%,特异度为86.67%,AUC面积为0.869;与对照组相比,LAMPS组细胞凋亡率、TNF-α、IL-1β、IL-6均显著升高[(8.24±1.35)%比(26.57±3.24)%、(7.23±2.12)pg/mL比(26.32±6.47)pg/mL、(11.62±3.21)pg/mL比(30.24±6.23)pg/mL、(11.20±3.21)pg/mL比(45.32±12.16)pg/mL,均P<0.05];与si-NC组相比,si-H19组细胞凋亡率、TNF-α、IL-1β、IL-6水平显著降低(P<0.05)。结论LncRNA H19在MPP病儿血清中高表达,并可抑制LAMPS诱导的肺泡巨噬细胞凋亡及炎症反应。

Objective To investigate the expression of long noncoding RNA H19(lncRNA H19)in the serum of children with My⁃coplasma pneumoniae pneumonia(MPP)and its effect on mycoplasma pneumoniae lipid-associated membrane protein(LAMPS)-in⁃duced alveolar macrophage apoptosis and inflammatory factor expression.Methods A total of 60 children with MPP who were admit⁃ted to Nanyang Central Hospital from October 2017 to December 2018 were collected as the observation group.According to the de⁃gree of the illness of the children,they were divided into convalescent stage(30 cases)and acute stage(30 cases).Sixty healthy chil⁃dren who underwent physical examination in our hospital during the same period were selected as the control group.Real-time quanti⁃tative polymerase chain reaction(qRT-PCR)was used to detect the expression level of LncRNA H19.A receiver operating characteris⁃tic(ROC)curve was used to analyze the diagnostic value of LncRNA H19 for MPP.The MH-S mouse alveolar macrophages were cul⁃tured in vitro,and the cells were treated with LAMPS to establish a cell injury model.si-NC and si-H19 were transfected into MH-S cells and treated with LAMPS for 24 h.Flow cytometry was used to detect the apoptosis rate.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6).Results Compared with the control group,the expression level of serum LncRNA H19 in the observation group was significantly increased[(1.01±0.13)vs.(2.58±0.25),P<0.05].Compared with the MPP recovery period,the expression level of serum LncRNA H19 in the acute phase of MPP was significantly increased[(1.63±0.20)vs.(3.24±0.39),P<0.05].ROC analysis showed that the sensitivity of ln⁃cRNA H19 in the diagnosis of MPP was 86.67%,the specificity was 86.67%,and the AUC was 0.869.Compared with the control group,the LAMPS group had a significantly increased apoptosis rate and TNF-α,IL-1βand IL-6 levels[(8.24±1.35)%vs.(26.57±3.24)%,(7.23±2.12)pg/mL vs.(26.32±6.47)pg/mL,(11.62±3.21)pg/mL vs.(30.24±6.23)pg/mL,(11.20±3.21)vs.(45.32±12.16)pg/mL,P<0.05].Compared with the si-NC group,the apoptosis rate and TNF-α,IL-1βand IL-6 levels of the si-H19 group were signifi⁃cantly reduced(P<0.05).Conclusion LncRNA H19 is highly expressed in the serum of children with MPP,and can inhibit LAMPS-induced apoptosis and the inflammatory response of alveolar macrophages.