目的 探讨呼吸窘迫综合征(Respiratory distress syndrome,RDS)新生患儿血浆miR-NAs的差异表达及意义.方法 选取68例RDS新生患儿、15例健康新生儿为研究对象,随机各选取6例进行血浆测序,用反转录聚合酶链反应(Reverse transcription-polymerase chain reaction,RT-PCR)检测血浆 miRNAs表达.鉴定出差异表达最显著的两个 miRNAs进行后续研究,用RT-PCR检测RDS 发作期、出院时、出院后1个月时的miRNAs水平.用受试者工作特征曲线(Receiver operating characteristic curve,ROC)分析发作期时血浆miRNAs表达水平诊断RDS严重程度的价值.分析发作期时miRNAs表达水平与动脉血氧分压(Partial pressure of oxygen,PaO2)、二氧化碳分压(Partial pressure of carbon dioxide,PaCO2)、吸氧分数(Fraction of oxygen uptake,FiO2)、氧合指数(PaO2/FiO2)、肌酸激酶同工酶(Creatine kinase isoenzyme,CK-MB)、乳酸脱氢酶(Lactic dehydrogenase,LDH)、超氧化物歧化酶(Superoxide dismutase,SOD)、丙二醛(Malondialdehyde,MDA)水平的相关性.结果 测序结果显示,RDS新生患儿与健康新生儿血浆中共1 230种差异表达miRNAs,其中693种表达上调,537种表达下调.miRNA-200b及miR-NA-449差异表达倍数>20,是差异性表达最显著的miRNAs.出院后1个月时RDS新生患儿血浆miRNA-200b及miRNA-449的表达水平均低于发作期和出院时(P<0.05),且出院时水平低于发作期(P<0.05).血浆 miRNA-200b、miRNA-449诊断RDS的曲线下面积(Area under curve,AUC)分别为0.702、0.763,灵敏度分别为73.6%、76.9%,特异度分别为69.8%、72.8%.血浆miRNA-200b与miRNA-449相对表达水平分别与PaO2、PaO2/FiO2 呈负相关(P<0.05),与PaCO2、FiO2 呈正相关(P<0.05).miRNA-200b、miRNA-449分别与CK-MB、LDH、MDA呈正相关(P<0.05),与SOD呈负相关(P<0.05).结论 RDS新生患儿血浆miRNAs有明显异常表达,其中miRNA-200b、miRNA-449差异较为显著,二者在RDS新生患儿血浆中表达水平升高,且与呼吸功能损伤、心肌损伤和氧化应激损伤相关.
Differential expression and miRNAs in plasma of newborns with respiratory distress syndrome(RDS)and its clinical significance
Objective To explore the differential expression and significance of plasma miRNAs in neonatal children with respiratory distress syndrome(RDS).Methods 68 newborn infants with RDS and 15 healthy newborns were selected as study subjects.6 of them were randomly selected for plasma sequen-cing,and then reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expres-sion of miRNAs in the plasma.Identify the 2 miRNAs with the most significant differential expression for subsequent research,and use RT-PCR to detect the levels of miRNAs during the onset of RDS,at dis-charge,and 1 month after discharge.The value of using Receiver Operating Characteristic Curve(ROC)to analyze plasma miRNAs expression levels during the seizure period in diagnosing the severity of RDS.Analyze the expression levels of miRNAs during the onset of RDS,as well as their correlation with the levels of partial pressure of oxygen(PaO2),partial pressure of carbon dioxide(PaCO2),fraction of oxy-gen uptake(FiO2),oxygenation index(PaO2/FiO2),serum creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA).Results The sequen-cing results showed that there were 1 230 differentially expressed miRNAs in the plasma of RDS newborns and healthy newborns,of which 693 were upregulated and 537 were downregulated.miRNA-200b and miRNA-449 have differential expression multiples greater than 20,making them the most significantly dif-ferentially expressed miRNAs.At 1 month after discharge,the expression levels of plasma miRNA-200b and miRNA-449 in newly born RDS patients were lower than those during the onset and discharge periods(P<0.05),and the levels at discharge were lower than those during the onset period(P<0.05).The area under curve(AUC)of miRNA-200b and miRNA-449 for diagnosing RDS were 0.702 and 0.763,re-spectively,with sensitivity of 73.6%and 76.9%,specificity of 69.8%and 72.8%.The relative expres-sion levels of plasma miRNA-200b and miRNA-449 were negatively correlated with PaO2 and PaO2/FiO2,respectively(P<0.05),while it positively correlated with PaCO2 and FiO2,respectively(P<0.05).MiRNA-200b and miRNA-449 were positively correlated with CK-MB,LDH and MDA(P<0.05),while it negatively correlated with SOD(P<0.05).Conclusion There is a significant abnormal expression of miRNAs in the plasma of newborn children with RDS,with significant differences in miRNA-200b and miRNA-449.The expression levels of the 2 are elevated in the plasma of newborn children with RDS and are associated with respiratory function damage,myocardial damage and oxidative stress damage.