Ningxue shengban decoction regulates T-cell immune balance in immune thrombocytopenia via the bone marrow hematopoietic microenvironment.

BACKGROUND

Immune thrombocytopenia (ITP) is characterized by increased platelet clearance and decreased platelet production. Hematopoietic stem and progenitor cell (HSPCs) abnormalities are a key mechanism of ITP, contributing to megakaryocyte defects and aberrant lymphocyte differentiation. Ningxue Shengban Decoction (NXSBD) has proven to be an effective therapeutic option for ITP.

AIM: This study aimed to explore the therapeutic mechanism of NXSBD in ITP.

METHOD: First, we evaluated the therapeutic effect of NXSBD on thrombocytopenia. T-lymphocyte subsets were analyzed by flow cytometry, megakaryocyte features were examined by HE staining, serum cytokines were quantified by ELISA, and hepatic/renal safety indices were measured by MS. Following the identification of core targets and pathways via network pharmacology, their expression in bone marrow cells was confirmed by single-cell RNA sequencing analysis. Pseudo-temporal analysis then tracked the dynamics of these targets during HSC lineage commitment, and molecular docking finally confirmed strong binding affinities with NXSBD's constituents.

RESULTS

NXSBD significantly ameliorated thrombocytopenia in ITP mice by rebalancing T-cell subsets and modulating key inflammatory cytokines, while also promoting the generation of thrombocytogenic megakaryocytes in the bone marrow. The core targets of NXSBD (RELA, IKBKB, AKT1, TP53, MAPK1, JUN, and FOS) were found to be present in hematopoietic stem cells (HSCs) and were involved in HSC differentiation into megakaryocytes and T lymphocytes. Molecular docking confirmed strong binding affinities between NXSBD constituents and these core targets. In conclusion, our findings demonstrate that NXSBD alleviates ITP through a multi-target mechanism that may participate in megakaryocyte production from HSCs and contribute to the restoration of peripheral T-cell homeostasis.