Analysis of Differentially Expressed Genes in the Peripheral Blood Transcriptome of Patients with High-Altitude Pulmonary Hypertension

HUANG Yixuan; YANG Feng; JIA Pengcheng; WANG Rui; LI Wu

doi:10.3639/j.issn.2095-3097.2025.12.010

Abstract：Objective This study employed RNA sequencing technology to analyze the peripheral blood of patients with high-altitude pulmonary hypertension(HAPH),aiming to identify core genes associated with HAPH's pathogenesis and to validate the expression changes of these genes using reverse transcription quantitative polymerase chain reaction(RT-qPCR).The study provides a theoretical basis for early diagnosis and the development of therapeutic targets for HAPH.Methods A total of 42 healthy males who underwent physical examinations at the 948th Hospital of the PLA(affiliated with the General Hospital of Xinjiang Military Region)from April to September 2023 and showed no abnormalities in cardiac ultrasound examinations were selected as study subjects.Following 5 months of high-altitude hypoxic exposure at elevations above 5 000 meters,they were divided into a healthy control group(24 cases)and an HAPH group(18 cases)based on Doppler ultrasound examinations.Blood samples were collected from both groups,and total RNA was extracted using TRIzol reagent.RNA sequencing was performed on the Illumina Novaseq 6000 platform.Differentially expressed genes(DEG)were identified using the limma package with thresholds of|log2FC|>1 and P<0.05.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was further conducted to identify pathways associated with the DEG.Protein-protein interaction(PPI)networks were constructed using the STRING database,and core genes were screened using the Maximum Neighborhood Component,Edge Percolated Component,and BottleNeck algorithms.The Xsum algorithm combined with the Cmap database was used to screen for potential compounds for HAPH treatment.Finally,RT-qPCR was performed to validate the expression changes of the core genes.Results Transcriptome sequencing identified 617 differentially expressed genes,including 326 upregulated and 291 downregulated genes.Pathway enrichment analysis revealed that the heme biosynthesis pathway was closely associated with HAPH.Based on PPI analysis,five core genes were identified:ferrochelatase(FECH),synuclein alpha(SNCA),actin beta,AKT serine/threonine kinase 1,and ubiquitin C.Screening via the Cmap database suggested arachidonyltrifluoromethane(ATFMK)as a potential compound that may inhibit the core genes.RT-qPCR validation results showed that FECH and SNCA were significantly upregulated in the HAPH group,indicating that these two genes may play important roles in the pathological process of HAPH.Conclusion This study identified differentially expressed core genes associated with HAPH through transcriptomic analysis.Among them,FECH and SNCA were further validated by RT-qPCR and may serve as potential biomarkers and therapeutic targets for HAPH.The compound ATFMK may have potential as a therapeutic agent.These findings provide new molecular insights for early diagnosis and targeted therapy of HAPH.