文献类型：期刊文献

英文题名：Integrating network pharmacology and proteomics to elucidate the mechanism of Allium macrostemon total saponins in alleviating high-altitude acute lung injury

作者：Liang, Keke[1,2];Yang, Jun[1,5];Wang, Renjie[4];Luo, Kai[1];Ma, Shuhe[1,2];Dou, Xia[1];Wang, Yihao[2];Gao, Yue[2,3];Zhang, Xianxie[1,2];Li, Maoxing[1,2,3]

第一作者：Liang, Keke

通信作者：Zhang, XX[1];Li, MX[1];Zhang, XX[2];Li, MX[2]

机构：[1]Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China;[2]Beijing Inst Radiat Med, Beijing 100850, Peoples R China;[3]Natl Key Lab Kidney Dis, Beijing 100850, Peoples R China;[4]Tianjin Univ Tradit Chinese Med, Tianjin 300193, Peoples R China;[5]Drug Safety Tech Inspect Ctr Ningxia Hui Autonomou, Yinchuan 750000, Peoples R China

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

通信机构：[1]corresponding author), Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China;[2]corresponding author), Beijing Inst Radiat Med, Beijing 100850, Peoples R China.|[1073501e14fb35863569f]甘肃中医药大学药学院（西北中藏药协同创新中心办公室）;[10735]甘肃中医药大学;

年份：2026

卷号：162

外文期刊名：PHYTOMEDICINE

收录：;Scopus(收录号：2-s2.0-105050529325);WOS:【SCI-EXPANDED(收录号:WOS:001882069800001)】；

基金：This study was supported by the "Innovation Star" Project for Outstanding Postgraduates of the Department of Education of Gansu Province (Grant number: 2026CXZX-922), the Outstanding Doctoral Student Project of Gansu Province (Grant number: 25JRRA258), the National Natural Science Foundation of China (Grant number: 82675327), and the Key Research and Development and Transformation Key Special Project of the Tibet Autonomous Region Science and Technology Plan (Grant number: XZ202601ZY0218).

语种：英文

外文关键词：Allium macrostemon Bunge total saponins (AMBS); High-altitude lung injury; JAK2/STAT3 signaling pathway; Malate dehydrogenase 2

摘要：Background High-altitude acute lung injury (HALI) is a severe respiratory disorder occurring after exposure to high-altitude hypoxic environments. Its pathological features primarily include pulmonary edema, inflammatory response, oxidative stress, and apoptosis, which may progress to high-altitude pulmonary edema in severe cases, posing a life-threatening risk. Allium macrostemon Bunge total saponins (AMBS), one of the main active components of A. macrostemon, have been investigated; however, their specific role and molecular mechanisms in HALI remain unclear. Purpose This study employed network pharmacology and proteomics approaches to investigate the protective effects and molecular mechanisms of AMBS against HALI. Methods Hypoxia models in mice were established to evaluate the anti-hypoxic activity and dose-response relationship of AMBS. HALI rat and mouse models were constructed using a simulated high-altitude hypobaric hypoxia chamber at 6500 m altitude. Parameters measured included lung index, lung wet/dry weight ratio, hematological parameters, inflammatory factors, oxidative stress markers, and apoptosis levels. Pathological changes in lung tissue were observed via hematoxylin and eosin (HE) staining. In vitro, a cobalt chloride (CoCl2)-induced hypoxic injury model in human pulmonary microvascular endothelial cells (HPMECs) and a 1% O-2 hypoxia model were established to assess cell viability, lactate (LACT) and lactate dehydrogenase (LDH) release, ROS levels, and mitochondrial membrane potential. Meanwhile, the contents of malate and oxaloacetate, as well as the activity and expression level of malate dehydrogenase (MDH), were measured in both in vivo and in vitro models. The mechanisms underlying AMBS-mediated improvement of HALI were elucidated using integrated network pharmacology and proteomics analysis, combined with surface plasmon resonance (SPR), RT-qPCR, Western blot, immunofluorescence, immunohistochemistry, and co-immunoprecipitation (Co-IP). Results AMBS significantly prolonged the survival time of hypoxic mice, reduced the lung index and lung wet/dry weight ratio in HALI rats, improved blood oxygen saturation and partial pressure of oxygen, corrected acid-base balance disturbances, and alleviated lung tissue pathological injury. Furthermore, AMBS decreased the levels of interleukin-1 beta (IL-1 beta), IL-6, tumor necrosis factor-alpha (TNF-alpha), cysteine-aspartic acid protease 3 (Caspase-3), and Bcl-2-associated X protein (Bax), while increasing glutathione peroxidase (GSH-Px), catalase (CAT), B-cell lymphoma 2 (Bcl-2), and superoxide dismutase (SOD) activities. In vitro experiments demonstrated that AMBS attenuated CoCl2-and 1% O-2-induced hypoxic injury in HPMECs by inhibiting LACT and LDH release, reducing ROS accumulation, and restoring mitochondrial membrane potential. In both HALI rats and HPMECs hypoxia models, hypoxia significantly decreased malate content while markedly increasing MDH activity and oxaloacetate content, and AMBS intervention reversed these metabolic abnormalities. Integrated network pharmacology and proteomics analysis revealed that AMBS alleviates HALI primarily through inhibition of the Janus kinase 2/signal transducer and activator of transcription 3/malate dehydrogenase 2 (JAK2/STAT3/MDH2) pathway. SPR confirmed that Macrostemonoside I, an active component of AMBS, directly binds to JAK2 and STAT3 proteins. WP1066 (a JAK2 inhibitor) produced protective effects consistent with AMBS, whereas STAT3 overexpression enhanced the protein-protein interaction between STAT3 and MDH2, antagonized the regulatory effects of AMBS on MDH2 expression and malate-oxaloacetate metabolic balance, and subsequently abolished the cytoprotective effects of AMBS. Conclusion AMBS likely exerts its protective effects against HALI by downregulating MDH2 expression through inhibition of the JAK2/STAT3 signaling pathway.