Treatment with Biaoxu Ganmao Granules ameliorates cyclophosphamide-induced immunosuppression in mice: Paeoniflorin-associated modulation of the TLR4/NF-κB pathway.

Objectives

Biaoxu Ganmao Granules (BXGG) is used clinically for wind-cold syndrome; however, its effects on cyclophosphamide (CTX)-induced immunosuppression and the associated pharmacological basis remain incompletely characterized.

Methods

Two mouse cohorts were used to evaluate BXGG and paeoniflorin in a CTX-induced immunosuppression model. Body weight, thymus and spleen indices, splenic histopathology, serum cytokines, and peripheral blood cell counts were assessed. Ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS), network pharmacology, and molecular docking were used to characterize BXGG constituents, blood-absorbed constituents, and putative targets. Paeoniflorin was selected as a representative blood-absorbed constituent for preliminary in vivo validation, and proteins related to the toll-like receptor 4/nuclear factor kappa B (TLR4/NF-κB) pathway were examined by Western blotting.

Results

BXGG partially reversed CTX-associated reductions in body weight, thymus and spleen indices. It also partially restored serum interleukin-2 (IL-2), interleukin-4 (IL-4), and interferon-gamma (IFN-γ) levels, and white blood cell (WBC), lymphocyte (LYMPH), platelet (PLT), and reticulocyte (RET) counts. Network analysis and docking prioritized paeoniflorin as a candidate immune-regulatory constituent. Paeoniflorin improved several CTX-impaired immune-related outcomes and decreased TLR4 expression and the phosphorylated inhibitor of κB alpha/inhibitor of κB alpha (p-IκBα/IκBα) and phosphorylated NF-κB p65/NF-κB p65 (p-p65/p65) ratios in spleen tissue.

Conclusions

BXGG ameliorated several CTX-induced immunosuppressive changes in mice. Paeoniflorin, selected as a representative absorbed constituent, partially reproduced the immunoprotective phenotype and was associated with attenuation of TLR4/NF-κB activation. The contributions and potential synergy of other absorbed compounds require further investigation.