支气管哮喘患儿长链非编码RNA PVT1 的表达 及与炎症因子的相关性

Expression of lncRNA PVT1 in children with bronchial asthma and its correlation with inflammatory factors

摘要:

目的 分析支气管哮喘患儿长链非编码RNA PVT1（lncRNA PVT1）的表达及与炎症因子的 相关性。方法 选取2019 年1 月—2019 年12 月张家口市第一医院收治的60 例支气管哮喘患儿作为观察 组，另选同期该院60 例健康体检儿童作为对照组。收集所有受试者血清和气管平滑肌细胞并进行原代培养， 运用RNA 过表达技术对对照组原代培养细胞进行lncRNA PVT1 过表达，采用聚合酶链反应检测lncRNA PVT1 表达水平；酶联免疫吸附试验测定两组原代培养细胞的炎症因子表达水平；用Pearson 法分析不同变 量的相关性；用受试者工作特征（ROC）曲线下面积（AUC）评价血清lncRNA PVT1 对小儿支气管哮喘的 诊断效能。结果 观察组血清及原代培养细胞lncRNA PVT1 表达水平高于对照组（P <0.05）。对照组过表 达lncRNA PVT1 后原代培养细胞中IL-1β、IL-6、IL-10 及TNF-α 表达水平较过表达前升高（P <0.05）。 观察组siRNA 沉默lncRNA PVT1 后原代培养细胞中IL-1β、IL-6、IL-10 及TNF-α 表达水平较沉默前 降低（P <0.05）。对照组过表达前后与观察组沉默前后原代培养细胞中IL-1β、IL-6、IL-10 及TNF-α 表 达水平的差值比较，差异无统计学意义（P >0.05）。经Pearson 法分析，原代培养细胞lncRNA PVT1 表达水 平与IL-1β、IL-6、IL-10 及TNF-α 表达水平均呈正相关（r =0.225、0.587、0.281 和0.312，P <0.05）。经 ROC 曲线分析，血清lncRNA PVT1 诊断小儿支气管哮喘的AUC 为0.917，标准误为0.074，敏感性为75.48% （95% CI ：0.856，0.954），特异性为94.03%（95% CI ：0.934，0.974）。结论 支气管哮喘患儿lncRNA PVT1 表 达水平较高，与炎症因子水平呈正相关，对该病的诊断效能较好，值得临床予以重视。

Abstract:

Objective To analyze the expression of lncRNA PVT1 in children with bronchial asthma and its relationship with inflammatory factors. Methods Sixty children with bronchial asthma treated in our hospital from January to December 2019 were selected as the observation group, and another 60 healthy children were selected as the control group. Serum and tracheal smooth muscle cells were collected from all subjects and primarily cultured. RNA overexpression technology was used to overexpress lncRNA PVT1 in the primarily cultured cells of the control group. Polymerase chain reaction (PCR) was used to detect the expression level of lncRNA PVT1, and the enzyme-linked immunosorbent assay (ELISA) was used to detect the expression level of inflammatory factors in primarily cultured cells before and after siRNA silencing of lncRNA PVT1. The relationship between different variables was analyzed using the Pearson method, while the area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the diagnostic efficacy of serum lncRNA PVT1 in children with bronchial asthma. Results The expression levels of lncRNA PVT1 in serum and primarily cultured cells in the observation group were higher than those in the control group (P < 0.05), while the expression levels of IL-1β, IL-6, IL-10 and TNF-α in primarily cultured cells after overexpression of lncRNA PVT1 in the control group were significantly higher than those before overexpression (P < 0.05). In the observation group, the expression levels of IL-1β, IL-6, IL-10, TNF-α in primarily cultured cells after lncRNA PVT1 silencing by siRNA were significantly lower than those before silencing (P < 0.05). There was no difference between the altered levels of IL-1β, IL-6, IL-10 and TNF-α in the primarily cultured cells before and after overexpression in the control group and those before and after silencing in the observation group (P > 0.05). As indicated by Pearson analysis, the expression level of lncRNA PVT1 in primarily cultured cells was positively correlated with the expression levels of IL-1β, IL-6, IL-10 and TNF-α (r =0.225, 0.587, 0.281, 0.312; P = 0.026, 0.000, 0.021, 0.013). The ROC curve analysis showed that the AUC of serum lncRNA PVT1 in the diagnosis of bronchial asthma in children was 0.917, with the standard error being 0.074, the sensitivity 75.48% (95% CI: 85.63%, 95.43%), and the specificity 94.03% (95% CI: 93.45%, 97.36%). Conclusions lncRNA PVT1 is highly expressed in children with bronchial asthma and has a positive correlation with the level of inflammatory factors. Thus, it is worthy of clinical application with great diagnostic efficacy.