Multi-omics analysis reveals the protective effects of Tibetan Medicine dalijuzhou powder on high-altitude polycythemia.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Da Li Ju Zhou (DLJZ) Powder is a traditional Tibetan medicinal compound preparation listed in the in the standards of the Chinese Ministry of Health. In recent years, DLJZ has been frequently used to treat high-altitude polycythemia (HAPC), though its mechanism of action and active ingredients remain unclear. AIM OF THE STUDY: This study aims to perform a comprehensive analysis of the chemical constituents of DLJZ, evaluate its efficacy against HAPC through multiple indicators, and investigate its action pathways and molecular mechanisms. MATERIALS AND METHODS: Chemical profiling of DLJZ was conducted using UPLC-MS and GC-MS. A rat model of HAPC was established through combined administration of CoCland exposure to hypobaric hypoxia. Multiple parameters, including complete blood count, blood gas analysis, coagulation tests, and hemorheology, were systematically evaluated to assess the therapeutic efficacy of DLJZ against HAPC. Untargeted metabolomics and Data-Independent Acquisition (DIA) proteomics were employed to identify key signaling pathways, which were subsequently validated by qPCR, ELISA, and IHC. RESULTS: DLJZ contained 191 bioactive compounds, comprising 132 non-volatile and 59 volatile components. In the HAPC rat model, DLJZ significantly reduced red blood cell count (p&#xa0;<&#xa0;0.01), hemoglobin concentration (p&#xa0;<&#xa0;0.01) and blood viscosity (p&#xa0;<&#xa0;0.01), improved alveolar ventilation, and normalized coagulation function. Mechanistically, DLJZ reduced erythropoiesis by downregulating the expression of EPO, HIF-1&#x3b1;, HIF-2&#x3b1;, PHD2, VHL, TFRC, and HMOX1 and upregulating FTH expression at both gene and protein levels. Additionally, DLJZ enhanced antioxidant defense by suppressing ALOX15 and promoting GPX4 expression, thereby inhibiting ferroptosis. CONCLUSION: DLJZ prevents the development of HAPC by modulating the hypoxia response and iron homeostasis through multiple molecular targets, thereby reducing excessive erythropoiesis. This study provides the first experimental evidence confirming the scientific basis of the traditional application of DLJZ, offering critical theoretical support for its clinical application and providing directions for future in-depth exploration. A limitation of this study is that only a single batch of DLJZ was used; therefore, the stability and batch-to-batch reproducibility of its effects require confirmation in future research.