背景:肺动脉高压(PAH)是一种进行性血管重塑疾病,其特征是持续的肺动脉压升高,会导致右心衰竭和过早死亡.鉴于PAH的发病机制复杂,预后差,生物标志物的鉴定和研究对于促进对该疾病的进一步了解变得越来越重要.
方法:与PAH相关的数据集,GSE49114、GSE180169和GSE154959是从公开的GEO数据库下载的。通过对GSE49114数据集执行WGCNA,共筛选出906个PAH相关关键模块基因。通过对GSE180169数据集进行差异分析,共鉴定出576个差异表达基因。此外,GSE154959单细胞测序数据集也进行了差异分析,导致内皮细胞内34个DEG的鉴定。通过将上述三组DEG相交,筛选出5个与PAH相关的hub基因,即Plvap,Cyp4b1、Foxf1、H2-Ab1和H2-Eb1,其中选择H2-Ab1用于随后的实验。
结果:使用SU5416/缺氧方法制备SuHx小鼠模型,并通过苏木精-伊红染色评价模型的成功构建,血液动力学检测,富尔顿指数,和西方印迹(WB)。WB和qRT-PCR的结果表明SuHx小鼠中H2-Ab1表达显著上调。与生物信息学分析的结果一致,在缺氧处理的小鼠肺动脉内皮细胞(PAECs)中,H2-Ab1表达呈时间依赖性增加.为了研究H2-Ab1是否影响PAH的发生和发展,我们下调了H2-Ab1在PAECs中的表达,发现它的击倒抑制了生存能力,附着力,迁移,和血管生成,同时促进PAECs的凋亡。
结论:H2-Ab1可以调节细胞增殖,凋亡,附着力,迁移,和PAECs的血管生成。
BACKGROUND: Pulmonary arterial hypertension (PAH) is a progressive disease of vascular remodeling characterized by persistent pulmonary arterial pressure elevation, which can lead to right heart failure and premature death. Given the complex pathogenesis and poor prognosis of PAH, the identification and investigation of biomarkers become increasingly critical for advancing further understanding of the disease.
METHODS: PAH-related datasets, GSE49114, GSE180169 and GSE154959, were downloaded from the publicly available GEO database. By performing WGCNA on the GSE49114 dataset, a total of 906 PAH-related key module genes were screened out. By carrying out differential analysis on the GSE180169 dataset, a total of 576 differentially expressed genes were identified. Additionally, the GSE154959 single-cell sequencing dataset was also subjected to differential analysis, leading to the identification of 34 DEGs within endothelial cells. By taking intersection of the above three groups of DEGs, five PAH-related hub genes were screened out, namely Plvap, Cyp4b1, Foxf1, H2-Ab1, and H2-Eb1, among which H2-Ab1 was selected for subsequent experiments.
RESULTS: A SuHx mouse model was prepared using the SU5416/hypoxia method, and the successful construction of the model was evaluated through Hematoxylin-Eosin staining, hemodynamic detection, fulton index, and Western Blot (WB). The results of WB and qRT-PCR demonstrated a significant upregulation of H2-Ab1 expression in SuHx mice. Consistent with the results of bioinformatics analysis, a time-dependent increase was observed in H2-Ab1 expression in hypoxia-treated mouse pulmonary artery endothelial cells (PAECs). To investigate whether H2-Ab1 affects the development and progression of PAH, we knocked down H2-Ab1 expression in PAECs, and found that its knockdown inhibited the viability, adhesion, migration, and angiogenesis, while concurrently promoted the apoptosis of PAECs.
CONCLUSIONS: H2-Ab1 could regulate the proliferation, apoptosis, adhesion, migration, and angiogenesis of PAECs.