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本研究旨在通过代谢组学筛选1,25-二羟基维生素D3[1,25-(OH)_2D3]预防低氧性肺损伤的生物标志物。将30只雄性SD大鼠(Rattus norvegicus)随机分为每组10只的对照(CA)组、低氧(HA)组和1,25-(OH)_2D3(MA)组, HA组大鼠在模拟海拔6 000 m低压氧舱内放置48 h,以构建低氧性肺损伤模型; MA组大鼠应用骨化三醇[1,25-(OH)_2D3]软胶囊溶于花生油中,每日按0.252μg/kg体重腹腔注射1次,连续注射5 d,随后同HA组大鼠一起在模拟海拔6 000 m低压氧舱内放置48 h; CA组大鼠饲养于当地海拔环境。造模结束后分别取3组大鼠的动脉血和肺组织。通过血气分析、肺组织干湿质量比方法,检测大鼠肺组织生理学和病理学变化。结果显示,与CA组相比, HA组动脉血氧饱和度(oxygen saturation, SaO2)和动脉血氧分压(oxygen partial pressure, PaO2)显著降低,肺组织含水量显著增加。与HA组相比, MA组SaO2、 PaO2显著升高,肺组织含水量显著降低。动脉血经离心收集上清液后,采用液相色谱-串联质谱技术(liquid chromatography-tandem mass spectrometry, LC-MS/MS)系统进行分析。以变量投影重要度(variable importance in the projection, VIP)大于1,学生t检验(student′s t test)P<0.05且倍数变化(fold change, FC)大于或等于1.2为条件初步筛选差异代谢物(differential metabolites, DEMs),使用京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)数据库查看DEMs通路富集情况。利用随机森林(random forest, RF)和支持向量机(support vector machine, SVM)对交集差异代谢物进行生物标志物筛选。经代谢组学分析筛选出4种潜在生物标志物,分别为D-果糖-1,6-二磷酸、 L-岩藻糖、尿刊酸和海藻糖。本研究初步筛选出1,25-(OH)_2D3预防低氧性肺损伤的潜在生物标志物,为其应用于低氧性肺损伤的临床治疗提供了参考依据。
Abstract：This study aims to screen the biomarker of 1,25-dihdroxyvitamin D3[1,25-(OH)_2D3] for the prevention of hypoxic lung injury based on metabolomics.Thirty male SD rats(Rattus norvegicus) were randomly assigned to three groups,including the control(CA) group,the hypoxia(HA) group,and the 1,25-(OH)_2D3(MA) group,with 10 animals in each group.To create a high-altitude hypoxia model,rats in the HA group were placed in a hypobaric chamber designed to simulate an altitude of 6 000 meters for 48 h.Rats in the MA group received calcitriol [1,25-(OH)_2D3] soft capsules dissolved in peanut oil,administered once daily via intraperitoneal injection at a dose of 0.252 μg/kg weight for 5 consecutive days.Following this,the rats were placed in the chamber with the HA group and exposed to 48 h of hypoxic stress.In contrast,the rats in the CA group were kept at the same altitude as their local environment.After model establishment,arterial blood and lung tissues from all three groups of animals were collected under normal altitude conditions.Physiological and pathological changes in lung tissues were assessed through blood gas analy sis,determination of lung tissue water content.The results showed that compared with the CA group,rats in the HA group had significantly increased lung tissue water content,along with significantly decreased oxygen saturation(SaO2) and partial pressure of oxygen partial pressure(PaO2).In contrast,the MA group showed significantly reduced lung tissue water content,and significantly improved SaO2 and PaO2 levels compared to the HA group.Arterial blood samples were collected from each group of rats for analysis.After centrifugation,the supernatant was introduced into the liquid chromatography-tandem mass spectrometry(LC-MS/MS) system for metabolite profiling of rats in each group.Differential metabolites(DEMs) were identified using the following criteria:variable importance in the projection(VIP)>1,Student's t test P<0.05,and fold change(FC)≥1.2.Pathway enrichment analysis of these DEMs was performed from kyoto encyclopedia of genes and genomes(KEGG) database.Biomarker screening for intersecting differential metabolism was conducted using two machine algorithms:random forest(RF) and support vector machine(SVM).Four potential biomarkers were screened out through metabolomics analysis,namely D-fructose 1,6-bisphosphate,L-fucose,urocanic acid,and trehalose.This study initially screened out potential biomarkers of 1,25-(OH)_2D3 for the prevention of hypoxic lung injury in rats,providing a reference basis for the clinical application of 1,25-(OH)_2D3 in the treatment of hypoxic lung injury.
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基本信息:
DOI：10.13417/j.gab.045.000481
中图分类号:R563
引用信息:
[1]程骞,苏红,戴重阳,等.基于代谢组学筛选1,25-(OH)_2D_3预防低氧性肺损伤的生物标志物[J].基因组学与应用生物学,2026,45(02):481-493.DOI:10.13417/j.gab.045.000481.
基金信息:
青海省中央引导地方科技发展资金项目(2025-ZY-044)资助
2026-01-14
2026-01-14
2026-01-14