Altered lncRNAs transcriptomic profiles in atherosclerosis-induced ischemic stroke. Altered lncRNAs transcriptomic profiles in atherosclerosis-induced ischemic stroke

Long non-coding RNAs (lncRNAs) can not only regulate gene transcription and translation, but also participate in the development of central nervous system diseases as epigenetic modification factors. However, their functional significance in atherosclerosis-induced ischemic stroke (AIIS) is unclear. The study aimed to screen out differentially expressed lncRNAs (delncRNAs), and to elucidate their potential regulatory mechanisms in the pathophysiology of AIIS. We screened out 10 patients with AIIS and recruited 10 healthy volunteers. We used microarray to detect the whole blood RNA of subjects, and explored the biological functions of delncRNAs by GO and KEGG analysis. After further analyzing the delncRNAs of THP-1 stimulated by ox-LDL, selective lncRNAs were screened. Through co-expression analysis, a corresponding lncRNA-mRNA interaction network was constructed. We yielded 180 delncRNAs (44 up-regulated and 136 down-regulated) and 218 demRNAs (45 up-regulated and 173 down-regulated). Lnc-SCARNA8 and lnc-SNRPN-2 are the highest elevated and decreased lncRNA in AIIS, respectively. The delncRNAs may play a significant role in ubiquitination-mediated protein degradation signaling pathways. In THP-1 cells models, the expression levels of lnc-SLC22A16-3, lnc-MTRNR2L1-10 and lnc-ACAT1-4 were consistent to the microarray results. According to lncRNA-mRNA network, the expression of vacuolar protein sorting 13 homolog B (VPS13B) and biliverdin reductase B (BLVRB) were significantly regulated. This study identified selective delncRNAs through microarray detection and validated by in vitro cell experiment. Our findings suggest that the ubiquitinated proteasome pathway, VPS13B and BLVRB may play a fundamental role in the pathological process of AIIS. Overall design: Based on the clinicopathological features and clinical images, we screened out 10 patients with AIIS and recruited 10 healthy volunteers. We used microarray to detect the whole blood RNA of subjects, and explored the biological functions of delncRNAs by GO and KEGG analysis. After further analyzing the delncRNAs of THP-1 stimulated by ox-LDL, selective lncRNAs were screened. Through co-expression analysis, a corresponding lncRNA-mRNA interaction network was constructed.

长链非编码RNA（Long non-coding RNAs，lncRNAs）不仅可调控基因的转录与翻译过程，还可作为表观遗传修饰因子参与中枢神经系统疾病的发生发展。然而，其在动脉粥样硬化性缺血性脑卒中（atherosclerosis-induced ischemic stroke，AIIS）中的功能意义尚不清楚。本研究旨在筛选差异表达长链非编码RNA（differentially expressed lncRNAs，delncRNAs），并阐明其在AIIS病理生理过程中的潜在调控机制。本研究纳入10例AIIS患者与10名健康志愿者，采用基因芯片技术检测受试者的全血RNA，并通过基因本体论（Gene Ontology，GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）富集分析探究delncRNAs的生物学功能。进一步分析氧化低密度脂蛋白（oxidized low-density lipoprotein，ox-LDL）刺激的THP-1细胞的delncRNAs后，筛选出目标lncRNAs，并通过共表达分析构建了相应的lncRNA-mRNA互作网络。本研究共获得180个delncRNAs（其中44个上调、136个下调）与218个差异表达mRNA（differentially expressed mRNAs，demRNAs，45个上调、173个下调）。Lnc-SCARNA8与Lnc-SNRPN-2分别为AIIS中表达上调最显著与下调最显著的lncRNAs。delncRNAs可能在泛素化介导的蛋白质降解信号通路中发挥重要作用。在THP-1细胞模型中，lnc-SLC22A16-3、lnc-MTRNR2L1-10与lnc-ACAT1-4的表达水平与基因芯片检测结果一致。基于lncRNA-mRNA互作网络，囊泡蛋白分选13同源物B（vacuolar protein sorting 13 homolog B，VPS13B）与胆绿素还原酶B（biliverdin reductase B，BLVRB）的表达均受到显著调控。本研究通过基因芯片检测筛选出目标delncRNAs，并通过体外细胞实验进行了验证。本研究结果表明，泛素化蛋白酶体通路、VPS13B与BLVRB可能在AIIS的病理进程中发挥关键作用。

实验整体设计：基于临床病理特征与临床影像资料，本研究纳入10例AIIS患者与10名健康志愿者，采用基因芯片技术检测受试者的全血RNA，并通过GO与KEGG分析探究delncRNAs的生物学功能。进一步分析ox-LDL刺激的THP-1细胞的delncRNAs后，筛选出目标lncRNAs，并通过共表达分析构建了相应的lncRNA-mRNA互作网络。