长链非编码RNA NONHSAT247814.1在18例儿童心肌炎中的表达及体外细胞实验观察

doi:10.6040/j.issn.1671-7554.0.2022.0152

摘要/Abstract

摘要： 目的探讨儿童心肌炎潜在的诊断标志物并明确其在人心肌细胞炎症中的生物学功能。方法筛选儿童心肌炎基因芯片结果中差异表达的长链非编码RNA(LncRNA),继而扩大样本量进行验证后,确立目标分子。收集心肌炎患儿18例(儿童心肌炎组)及健康查体儿童18例(正常对照组)的基本资料及临床信息,采用实时定量聚合酶链反应(qRT-PCR)法检测目标分子的表达水平,绘制受试者操作曲线(ROC)判断目标分子用于诊断儿童心肌炎的价值。利用脂多糖刺激人心肌细胞(AC16)构建心肌细胞炎症模型,将小干扰RNA转染至AC16中抑制目标分子的表达。采用酶联免疫吸附(ELISA)法检测细胞上清中炎症因子及心肌损伤标志物的蛋白浓度,采用CCK8法检测细胞的增殖活性。结果确定NONHSAT247814.1为本研究的目标分子。NONHSAT247814.1在儿童心肌炎组的相对表达量高于正常对照组(P<0.01),其在急性期的相对表达量高于恢复期。与外周循环中的N端脑钠肽前体呈正相关(R2=0.40, P=0.005),与左心室射血分数呈负相关(R2=-0.53, P=0.000 6); NONHSAT247814.1诊断儿童心肌炎的ROC曲线(AUC=0.89, P<0.001,灵敏度为83.33%,特异度为88.89%);在细胞水平,降低NONHSAT247814.1的表达,可以减少白介素-1β(IL-1β)、白介素-6(IL-6)、肌钙蛋白(cTnT)及B型利钠肽(BNP)的释放,增加细胞的增殖活性。结论 NONHSAT247814.1具有儿童心肌炎诊断标志物潜力,抑制NONHSAT247814.1的表达,可以减轻LPS诱导的心肌细胞炎症损伤。

关键词: 长链非编码RNA, NONHSAT247814.1, 心肌炎, 诊断标志物, 儿童

Abstract: Objective To explore the potential diagnostic marker of child myocarditis and to clarify the biological functions in human cardiomyocyte inflammation. Methods Long non-coding RNAs(LncRNAs)differentially expressed in the results of pediatric myocarditis gene chips were screened, the sample size was expanded for validation, and the target molecule was identified. The basic data and clinical information of 18 children with myocarditis(myocarditis group)and 18 healthy children(normal control group)were collected, real-time quantitative polymerase chain reaction(qRT-PCR)was used to test the expression of the target molecule, and then the receiver operating characteristic(ROC)curve was plotted to evaluate the value of the target molecule in the diagnosis of pediatric myocarditis. Lipopolysaccharide was utilized to stimulate AC16 cells and then extoblish the model of inflammatory injury, and the small interfering RNA was transfected into AC16 to inhibit the expression of the target molecule. Enzyme-linked immunosorbent assay(ELIAS)was used to test the protein concentration of inflammatory factors and myocardial injury markers in the cell supernatants, and CCK8 was used to detect cellular proliferation. Results NONHSAT247814.1 was identified as the target molecule. The relative expression of NONHSAT247814.1 was higher in the myocarditis group than in normal control group(P<0.001). The relative expression was higher in acute phase than in recovery phase, and positively correlated with NT-pro brain natriuretic peptide in the peripheral circulation(R2=0.40, P=0.005), but negatively correlated with left ventricular ejection fraction(R2=-0.53, P=0.0006). ROC for NONHSAT247814.1 in diagnosing myocarditis was(AUC=0.89, P<0.001, sensitivity=83.33% and specificity=88.89%). At the cellular level, reducing the expression of NONHSAT247814.1 attenuated the release of interleukin-1β, interleukin-6, troponin T and brain natriuretic peptide, while increased the cellular proliferation. Conclusion NONHSAT247814.1 has potential as a diognostic marker for myocarditis in children, inhibiting of NONHSAT247814.1 expression attenuates LPS-induced inflommation iujury in cardiomyocytes effects at the cellular level.

Key words: Long non-coding RNA, NONHSAT247814.1, Myocarditis, Diagnostic marker, Children

中图分类号:

R725.4

冯鑫鑫,韩波,张丽,马孟洁,陈思宇. 长链非编码RNA NONHSAT247814.1在18例儿童心肌炎中的表达及体外细胞实验观察[J]. 山东大学学报 (医学版), 2022, 60(10): 27-32.

FENG Xinxin, HAN Bo, ZHANG Li, MA Mengjie, CHEN Siyu. Long non-coding RNA NONHSAT247814.1 expression in 18 childhood with myocarditis and in vitro cellular experimental observation[J]. Journal of Shandong University (Health Sciences), 2022, 60(10): 27-32.

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