Network-based pharmacological and experimentally validated study on the therapeutic effects and mechanisms of Polygonatum Rhizoma Ginseng Formula in immunocompromised mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Polygonatum Rhizoma Ginseng Formula (HJRS) is a traditional Chinese medicine preparation commonly used to enhance immunity. Traditionally, it is believed to "moisten dryness and nourish yin, tonify the spleen, and benefit qi." However, a systematic evaluation of its efficacy and mechanism of action is still lacking.

AIM OF THE STUDY: To explore the pharmacodynamic effects and underlying mechanisms of HJRS in cyclophosphamide (CTX)-induced immunocompromised model mice.

METHOD: To explore the immunomodulatory effects of HJRS, we used cyberpharmacology combined with in vivo experiments. An immunocompromised mouse model was established using CTX, and different doses of HJRS were administered by gavage during the modeling period. The efficacy of HJRS was evaluated by assessing immune-related parameters, including body weight, heat pain threshold, and behavioral activity. Biochemical markers (acid phosphatase, total nitric oxide synthase, and lactate dehydrogenase), immune function indicators (C3, C4, secretory immunoglobulin A [sIgA]), and immune organ indices (spleen and thymus coefficients) were measured. Additionally, intestinal mucosal immunity (Peyer's patches, sIgA, and IgM), cellular immunity (delayed-type hypersensitivity [DTH]), and humoral immunity (serum hemolysin, hemolysin, and IgM) were analyzed in the model mice. To further investigate the underlying mechanisms, we used 16S RNA sequencing, western blotting, gas chromatography-mass spectrometry, and other techniques to examine the intestinal microbiota, the short-chain fatty acids-G protein-coupled receptor 41 (SCFAs-GPR41) pathway, and the downstream mitogen-activated protein kinase (MAPK) signaling pathway.

RESULT: Network pharmacology analysis identified 34 active compounds and predicted 154 potential targets in HJRS that may regulate immune function by interacting with multiple targets (tumor-necrosis factor [TNF], PIK3CG, JUN, AKT1, and MAPK) and immune-related signaling pathways (MAPK and TNF-signaling pathway). Experimental validation demonstrated that HJRS alleviated symptoms of weakness and reduced behavioral activity in the model mice. It significantly enhanced the DTH response, elevated serum hemolysin levels, and restored the balance of white blood cells, lymphocytes, red blood cells, and hemoglobin percentages. HJRS also rebalanced the serum CD3+CD4+/CD3+CD8+ ratio and increased C3 and C4 levels. Furthermore, it improved immune organ function by raising the thymus and spleen coefficients and increasing serum IgA, IgA, and IgM levels. Histopathological examination revealed that HJRS ameliorated CTX-induced pathological damage in the thymus, spleen, and intestines. HJRS regulated gut microbiota composition by enhancing the richness of Bacteroidetes and increasing the Firmicutes/Bacteroidetes ratio, while reducing the abundance of Firmicutes. It decreased the relative abundance of harmful genera, such as Bacteroides and Enterorhabdus, and promoted the growth of beneficial genera, including Muribaculaceae, Clostridia, and Alloprevotella. Additionally, HJRS increased the fecal acetic acid, propionic acid, hexanoic acid, and total SCFA levels in the model mice. It activated the SCFAs-GPR41 pathway and downstream MAPK signaling, thereby enhancing intestinal mucosal immune function.

CONCLUSION: HJRS effectively improved immune function in CTX-induced immunocompromised mice. Its mechanism of action may be associated with the regulation of intestinal microbiota dysbiosis, activation of the intestinal "SCFA-GPR41" pathway, and modulation of the downstream MAPK signaling pathway.