脂多糖调控对大鼠心脏微血管内皮细胞的转录组分析

Transcriptomic characteristics of lipopolysaccharide-induced cardiac microvascular endothelial cells

目的探索脂多糖(LPS)对大鼠心脏微血管内皮细胞(rCMECs)转录组的调节作用。方法对照组(正常培养rCMECs),LPS组(100 ng/mL LPS处理6 h的rCMECs),每组进行3个生物学重复转录组测序。得到差异基因后,使用实时定量PCR对部分差异基因mRNA的表达进行验证。分别对上调和下调基因进行GO和KEGG富集,并对差异基因进行共表达网络分析。采用独立t检验进行统计学分析。结果LPS处理后,265个基因表达上调,118个基因的表达下调。前10个最显著上调基因为:Mt2a、Cyp7b1、Sod2、Icam1、Ccl2、AC128848.1、Mt1、Cebpd、Serpinb2和Tnfrsf11b。前10个最显著下调基因为:Cavin2、Ankrd1、Edn1、Prss35、Lmod1、Dhrs3、Ttc22、Sema6a、Map2k3和Sema7a。定量PCR的结果表明Mt2a、Sod、Ccl2、Cxcl1、Icam1和Vcaml基因的表达得到了上调(P均<0.01);而Cavin2、Ankrd1、Edn1和Prss35基因表达下调(P均<0.05)。GO和KEGG富集的结果表明,上调基因与内皮细胞对炎性免疫细胞的趋化作用和黏附作用密切相关;而下调基因则是与钙离子信号和G蛋白相关通路以及内皮通透性增加有关。此外,差异基因进行共表达网络分析发现Sod2处于核心位置,提示其可能与LPS诱导的rCMECs的各种变化密切相关。结论LPS调控了rCMECs中大量与炎性免疫细胞进入心肌组织相关基因的表达。

Objective To explore the role of lipopolysaccharide(LPS)in regulating the transcriptome of rat cardiac microvascular endothelial cells(rCMECs).Methods Control group(rCMECs under normal culture condition),LPS group(rCMECs treated with 100 ng/mL LPS for 6 h),each group was subjected to 3 repeatly transcriptome sequencing.After identified differentially expressed genes,quantitative real-time PCR(q-PCR)was used to verify mRNA expression for some of differentially expressed genes.GO(Gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)enrichment were carried out in up-regulated and down-regulated genes respectively and co-expression network analysis was conducted fordifferentially expressed genes.Statistical analysis was performed using the unpaired t-test.Results 265 genes were up-regulated and 118 down-regulated after LPS treatment.The 10 most significant up-regulated genes were Mt2a,Cyp7b1,Sod2,Icam1,Ccl2,AC128848.1,Mt1,Cebpd,Serpinb2 and Tnfrsf11b.while the 10 most significant down-regulated genes were Cavin2,Ankrd1,Edn1,Prss35,Lmod1,Dhrs3,Ttc22,Sema6a,Map2k3 and Sema7a.The results of q-PCR showed that the expressions of Mt2a,Sod2,Ccl2,Cxcl1,Icam1 and Vcaml indeed up-regulated(P<0.01),and the expressions of Cavin2、Ankrd1、Edn1 and Prss35 were down regulated(P<0.05).The enrichment of GO and KEGG showed that up-regulated genes and inflammatory immune cellswere related tochemotaxis and adhesion of endothelial lated to calcium ion signal and G proteinsicells.The down-regulated were related to calcium signaling and G proteinrelated pathways as well as increased endothelial permeability.In addition,the co-expression network analysis of differentially expressed genes showed that Sod2 was the key gene,which suggested that it may be related to the changes of rCMECs induced by LPS.Conclusion Lipopolysaccharide regulates the large numbers of genes of inflammatory immune cells entering myocardial tissue in rCMECs.