As is known to all, the immune system is the most complex biological system. It is the foundation of the human defense system and has the ability to recognize and eliminate foreign pathogens (Li and Wu, 2021). Besides, it can harmonize with other systems in vivo to maintain the stability of the physical environment and physiological balance. When immune system is disrupted or suppressed, our bodies can develop severe inflammatory and immune disorders that seriously affect quality of life (Dorshkind et al., 2009).

Cyclophosphamide (CTX) is an extensively used chemotherapeutic drug in clinical practice. It is an important drug in the therapy of neoplastic diseases by exerting its anti-neoplastic effects through alkylation. Since it is a non-specific drug that acts on the cell cycle, it could also inhibit normal immune cells, finally resulting severe inflammation and immunosuppression (Emadi et al., 2009). However, there is still no good way to relieve immunosuppression caused by CTX. Therefore, it is necessary to seek out an appropriate drug to reduce the toxic of CTX.

Astragalus mongholicus Bunge (AM), a famous Chinese tonic medicinal herb which has been used for thousands in treating immune-mediated diseases and has similarly shown significant improvement in CTX-mediated immunosuppression in our previous studies (Liu et al., 2021). Astragaloside III (AS-III), an active triterpenoid saponin derived from AM, has been shown to have significant pharmacological effects in immune system activation such as enhancing anti-tumor reaction of NK cells (Chen et al., 2019). Nevertheless, the underlying protective mechanism of AS-III on CTX-induced immunosuppression is still unclear.

Network pharmacology is an emerging methodology based on systems biology theories that can be used to study therapeutic targets and biomarkers for complicated diseases (Nogales et al., 2022; Xu, J. et al., 2023). By integrating the multi-level data of “component-target-pathway-disease”, the complex prevention and treatment mechanisms of traditional Chinese medicine (TCM) can be elucidated (Wang et al., 2021). Molecular docking technology is an important part of the drug discovery and it theoretically provides the possible modes of action between drugs and their related proteins (Stanzione et al., 2021).

The experiments in this article aim to research the regulation mechanism of AS-III in antagonizing effect of immunosuppression on the basis of the comprehensive utilization of network pharmacology, molecular docking, together with in vitro/vivo experiments verification. Finally, all the experimental data of biological indicators, relative cytokine secretion, mRNA and protein expression have confirmed our theoretical analysis. These findings provide an experimental foundation for the use of AS-III as a potential drug candidate for the management of immunosuppression.