血清外泌体LncRNA作为强直性脊柱炎早期诊断分子标记物的筛选研究

Screening Study on Serum Exosomal LncRNA as Early Diagnostic Molecular Markers for Ankylosing Spondylitis

目的:筛选血清外泌体LncRNA作为强直性脊柱炎(AS)早期诊断分子标记物。方法:收集了本院2021年1月至2022年12月医院的AS患者的全血样本,通过超速离心从AS患者(n=50)和健康个体(n=50)的血浆中分离外泌体,使用TRIzol试剂提取总外泌体RNA,采用微阵列分析法对两组患者进行LncRNA分析,实时定量逆转录-聚合酶链反应(qRT-PCR)测定LncRNA在所有患者和健康受试者中的表达水平。结果:本研究结果表明通过qRT-PCR发现6种lncRNA(LNCV6_116109、LNCV6_98390、LNCV6_38772、LNCV6_108266、LNCV6_84003和LNCV6_98602)在AS患者中的表达显著上调,而不是健康个体。采用受试者工作特性曲线(ROC)验证其诊断准确性。lncRNA的曲线下面积值分别为0.8052(LNCV6_116109)、0.7088(LNCV6_98390)、0.7460(LNCV6_38772)、0.7292(LNCV6_108266)、0.7356(LNCV6_84003)和0.6800(LNCV6_98602)。结论:本研究表明外泌体LncRNA(LNCV6_84003、LNCV6_98602、LNCV6_108266、LNCV6_116109、LNCV6_98390和LNCV6_38772)的表达在AS患者的血浆中显着上调,并且它们可能作为早期诊断AS的潜在非侵入性生物标志物。

Objective:To screen serum exosomal long non-coding RNA(LncRNA)as potential molecular markers for the early diagnosis of ankylosing spondylitis(AS).Methods:Whole blood samples of AS patients in our hospital from January 2021 to December 2022 were collected.Exosomes were isolated from the plasma of AS patients(n=50)and healthy individuals(n=50)by ultrafast centrifuge.Total exosomal RNA was extracted using TRIzol reagent.LncRNAs were analyzed by microarray analysis in the two groups of patients,and the expression levels of LncRNAs in all patients and healthy subjects were measured by real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Results:The results of this study showed that six lncrnas(LNCV6_116109、LNCV6_98390、LNCV6_38772、LNCV6_108266、LNCV6_84003和LNCV6_98602)were significantly up-regulated in AS patients by qRT-PCR.Not a healthy individual.Receiver operating characteristic curve(ROC)was used to verify its diagnostic accuracy.The area values under the curve of lncRNA were 0.8052(LNCV6_116109)、0.7088(LNCV6_98390)、0.7460(LNCV6_38772)、0.7292(LNCV6_108266)、0.7356(LNCV6_84003)and 0.6800(LNCV6_98602).Conclusion:This study showed that the expression of exosome LncRNAs(LNCV6_84003,LNCV6_98602,LNCV6_108266,LNCV6_116109,LNCV6_98390 and LNCV6_38772)was significantly up-regulated in plasma of patients with AS.They may serve AS potential non-invasive biomarkers for early diagnosis of AS.