LPS诱导的急性肺损伤中早期标志物表达研究

Expression of LncRNA in Endotoxin-induced Acute Lung Injury in Early Stage and Its Relationship with Lung Injury

目的探讨长链非编码RNA在内毒素诱导的急性肺损伤早期表达。方法应用脂多糖(Lipopolysacharide,LPS)在健康昆明小鼠气管内滴注建立急性肺损伤动物模型;查找文献,筛选小鼠肺组织在炎症后差异表达的lnc RNAs,选取部分lnc RNAs(LncRNA IL-7R、LncRNA H19、LncRNAHotair、LncRNA Maltal1、LncRNA acta-1、LncRNA Cot2)进行实时荧光定量PCR验证和生物信息学分析,验证可能与LPS诱导的急性肺损伤发生相关的lnc RNAs表达的变化,及其与LPS诱导急性肺损伤主要信号通路TLR4-MYD88-NF-kb P65通路的相关成员的变化水平,为其在急性肺损伤发生中的机制研究提供前期基础。结果气道内直接滴入LPS 24h后,TLR4、MYD88、LncRNAH19在LPS诱导的急性肺损伤小鼠模型的肺组织中,实验组较对照组的基因表达水平增高(P<0.05);LncRNA IL-7R、LncRNAHotair、LncRNA Maltal1、LncRNA acta-1、LncRNA Cox2基因水平的表达,两组比较无明显差异性(P>0.05)。经地塞米松干预后,在急性肺损伤炎症小鼠肺组织中,干预组TLR4、DYD88、LncRNA H19的基因表达水平均较实验组出现不同程度降低(P<0.05)。结论 LncRNA H19、LncRNA IL-7R可能通过信号通路TLR4-MYD88-NF-kb P65参与内毒素诱导的急性肺损伤中早期表达,但其与肺损伤的关系有待进一步研究。

Objective To investigate the early expression of long-chain non-coding RNA in endotoxin-induced acute lung injury. Methods Lipopolysacharide （LPS） was used to establish animal model of acute lung injury in Kunming mice by tracheal instillation.After searching the literature,lnc RNAs differentially expressed in the lung tissues of mice were screened,and some lnc RNAs （LncRNA IL-7R,LncRNA H19,LncRNA Hotair,LncRNA Maltal1,LncRNA acta-1,LncRNA Cox2） were detected by real-time fluorescence quantitative PCR and bioinformatics analysis to verify the changes of lnc RNAs expression possibly related to LPS-induced acute lung injury,LPS-induced changes in the level of TLR4-MYD88-NF-kb P65 pathway in acute lung injury and provide a preliminary basis for its mechanism in the pathogenesis of acute lung injury. Results After direct intratracheal instillation of LPS for 24 hours,TLR4,MYD88,and LncRNA H19 in the LPS-induced acute lung injury mouse model,the expression level of the experimental group was higher than that of the control group （ P 〈0.05）;LncRNA IL-7R Expression levels of LncRNA Hotair,LncRNA Maltal1,LncRNA acta-1,and LncRNA Cox2 genes were not significantly different between the experimental group and the control group （ P 〉0.05）.Dexamethasone intervention was used to inhibit the inflammatory process of acute lung injury.In mouse lung tissue,the expression levels of TLR4,DYD88,and LncRNA H19 genes in the intervention group were lower than the experimental group.There were different degrees of reduction （ P 〈0.05）. Conclusion LncRNA H19 and LncRNA IL-7R may be involved in the early expression of LPS-induced acute lung injury through the signaling pathway TLR4-MYD88-NF-kb P65,and its relationship with lung injury needs further study.