基于非靶向代谢组学的鸟嘌呤核苷干预OVA诱导过敏性哮喘小鼠的作用及机制研究

Study on the effects and mechanisms of guanosine intervention on OVA-induced allergic asthma in mice based on untargeted metabolomics

关键词:  鸟嘌呤核苷  过敏性哮喘  代谢组学  免疫调节  抗炎作用

Key words:  guanosine  allergic asthma  metabolomics  immunomodulation  anti-inflammatory effect

摘要:

目的 本研究旨在通过代谢组学分析，探讨鸟嘌呤核苷对过敏性哮喘小鼠的干预作用及其潜在机制，特别关注其在宿主免疫反应和代谢调节中的作用，为哮喘的临床治疗提供理论依据。方法 研究使用40只SPF级雄性Balb/c小鼠，随机分为正常对照组（NC）、过敏性哮喘模型组（M）、阳性药组（孟鲁司特钠，Y, 1.3 mg/kg）、鸟嘌呤核苷-低组（Guanosine-L, 15 mg/kg）和鸟嘌呤核苷-高组（Guanosine-H, 30 mg/kg），具体范围基于前期预实验结果。通过卵清蛋白（OVA）诱导建立过敏性哮喘小鼠模型。连续给药7天后，检测小鼠咳喘指标、肺功能、肺系数，苏木精-伊红（HE）染色观察小鼠肺组织病理变化，PAS染色观察粘液分泌，马松（MASSON）染色观察其胶原纤维沉积水平，酶联免疫吸附测定（ELISA）小鼠血清和肺泡灌洗液中炎症因子水平，组织免疫荧光检测免疫细胞水平，并采用液质联用技术（LC-MS）对小分子代谢产物进行分析，探究特征代谢物所参与的生物代谢途径。结果 与模型组相比，Guanosine-H组小鼠的肺组织病理损伤显著改善，尤其在气道炎症和肺泡结构修复方面改善幅度相对更大，差异具有统计学意义（P＜0.05）。免疫细胞分析表明，鸟嘌呤核苷显著降低了肺组织中嗜酸性粒细胞、Th2细胞和巨噬细胞的浸润，并抑制了相关细胞因子的分泌（P＜0.05）。代谢组学分析显示，鸟嘌呤核苷通过调节氨基酸代谢、脂质代谢等关键通路，显著改善了由OVA引起的代谢物差异。尤其在Guanosine-H组（高剂量组），在这些代谢通路的调节上表现出更为显著的效果（P＜0.05）。结论 鸟嘌呤核苷通过调节免疫细胞浸润、改善肺组织病理结构以及调控关键代谢通路（如氨基酸代谢、脂质代谢等），在过敏性哮喘小鼠模型中，在本实验条件下显示出抗哮喘样效应（P＜0.05）。本研究为鸟嘌呤核苷作为抗过敏性哮喘的新疗法提供了实验理论依据，并为其未来的临床应用奠定了实验基础。

Abstract:

OBJECTIVE The purpose of this study is to explore the anti asthma effects and potential mechanisms of guanosine on allergic asthma mouse models through metabolomics analysis, with a particular focus on its role in host immune response and metabolic regulation. METHODS The study utilized 40 SPF-grade male Balb/c mice, randomly assigned to a normal control group (NC), an allergic asthma model group (M), 阳性药组 drug group (Montelukast sodium, Y, 1.3 mg/kg), low-dose guanosine nucleoside group (Guanosine-L, 15 mg/kg), and high-dose guanosine nucleoside group (Guanosine-H, 30 mg/kg). Dosage ranges were selected based on preliminary pre-experimental results. An ovalbumin (OVA)-induced allergic asthma mouse model was established. After 7 consecutive days of drug administration, coughing and wheezing indices, lung function, and lung coefficients were measured. Hematoxylin-eosin (HE) staining assessed pulmonary histopathology, PAS staining evaluated mucus secretion, Masson's trichrome staining examined collagen fiber deposition levels, Enzyme-linked immunosorbent assay (ELISA) was used to measure inflammatory cytokine levels in serum and bronchoalveolar lavage fluid. Tissue immunofluorescence assessed immune cell levels, while liquid chromatography-mass spectrometry (LC-MS) analyzed small-molecule metabolites to investigate the biological pathways involving characteristic metabolites. RESULTS Compared with the model group, mice in the Guanosine-H group showed significantly improved pulmonary tissue pathology, with particularly greater and statistically significant improvements in airway inflammation and alveolar structural repair (P < 0.05). Immune cell analysis revealed that guanosine nucleoside significantly reduced the infiltration of eosinophils, Th2 cells, and macrophages in lung tissue while suppressing the secretion of related cytokines. Metabolomic analysis revealed that guanosine nucleoside significantly improved OVA-induced metabolic dysregulation by modulating key pathways including amino acid metabolism and lipid metabolism. Notably, the Guanosine-H group (high-dose group) exhibited more pronounced effects in regulating these metabolic pathways（P＜0.05）.CONCLUSION In the mouse model of allergic asthma under the experimental conditions, guanine nucleoside demonstrated anti-asthmatic effects (P < 0.05) by regulating immune cell infiltration, improving pulmonary tissue pathology, and modulating key metabolic pathways such as amino acid and lipid metabolism. This study provides experimental theoretical support for guanine nucleoside as a novel anti-allergic asthma therapy and lays an experimental foundation for its future clinical application. KEY WORDS: guanosine; allergic asthma; metabolomics; immunomodulation; anti-inflammatory effect.