System pharmacology reveals the mechanism of action of Shengbai Oral Liquid alleviates chemotherapy-induced leukopenia by interfering the pro-platelet basic protein: Integrated network study of multi-omics.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium brevicornu Maxim., when used alone or as the principal component in traditional Chinese medicine (TCM) formulas, has the function of restoring internal "organs" and relieving fatigue. Shengbai Oral Liquid (SBOL), a TCM formula primarily composed of Epimedium brevicornu Maxim., has demonstrated clinical efficacy in managing chemotherapy- or radiotherapy-induced leukocytopenia. Nevertheless, its therapeutic mechanisms remain unclear.

AIM OF THE STUDY: Elucidate the mechanism of SBOL in treating chemotherapy-induced leukopenia.

METHODS

A leukopenia model in mice was established using cyclophosphamide (CTX). Transcriptomic, proteomic, and metabolomic studies were performed to investigate mechanistic effects of SBOL at the genetic, protein, and metabolic levels. A comprehensive approach integrating multi-omics and network pharmacology was then employed to predict potential targets and pathways underlying the effects of SBOL. Finally, experiments were performed to validate the underlying molecular mechanisms and identify potential therapeutic targets.

RESULTS

SBOL (oral treatment for 18 days) significantly increased the white blood cell (WBC) count in the peripheral blood of mice with leukopenia. Additionally, SBOL can significantly increase the quantity and proliferative activity of bone marrow cells (BMCs). Further flow cytometry analysis showed that SBOL can increase the number of hematopoietic stem and progenitor cells (HSPCs) in BMCs. Specifically, it increased the populations of hematopoietic stem cells (HSCs), hematopoietic progenitor cells (HPCs), short term hematopoietic stem cells (ST-HSCs), and multipotent progenitor cells (MPPs) within the HSPCs. What is more, SBOL can also reduce spleen index and reverse the compensatory increase of HSPCs in the spleen. Transcriptomic analysis indicated SBOL is involved in lipid metabolism and immune regulation, while proteomic analysis identified pathways related to leukocyte migration and chemotaxis. Metabolomic analysis highlighted arachidonic acid and amino acid metabolism. Multi-omics and network pharmacology studies have suggested that Pro-Platelet Basic Protein (PPBP), ALOXE3, and CCR1 are key targets. Experiments confirmed that SBOL significantly elevated levels of hematopoietic factors, including CXCL12, TGF-β1, IL-34, IL-5, and GM-CSF. Additionally, SBOL reversed the decreased expression of PPBP in BMCs and restored serum PPBP levels in these mice with leukopenia.

CONCLUSION

SBOL significantly alleviates leukopenia induced by chemotherapy. It increases the bone marrow HSPCs counts and elevates serum hematopoietic factors levels in chemotherapy-treated mice, thereby restoring hematopoietic function and boosting WBCs counts. PPBP is closely related to the treatment of chemotherapy induced leukopenia by SBOL.