Multi-omics reveals the mechanism of Rehmannia glutinosa Libosch. Processed with Amomum villosum Lour. in treating blood deficiency syndrome.

Amomi Fructus (Amomum villosum Lour.)-processed Rehmanniae Radix (Rehmannia glutinosa Libosch.) is a traditional Chinese herbal preparation used to tonify blood and improve digestive tolerance. It is considered a rational processing method to enhance the efficacy of Rehmanniae Radix (RR) in blood deficiency syndrome (BDS). This study investigated the therapeutic effects of Amomi Fructus -processed Rehmanniae Radix (AR) on BDS and explored its underlying mechanisms using a multi-omics approach. A rat model of BDS was established using cyclophosphamide and acetylphenylhydrazine. Rats were treated with AR or RR. Hematological parameters, spleen histopathology, TUNEL staining, bone marrow apoptosis by flow cytometry, transcriptomics, spleen metabolomics, and gut microbiota profiling were performed. AR significantly improved RBC, WBC, HGB, and HCT levels and alleviated spleen pathological injury in BDS rats. Transcriptomic analysis showed that the PI3K/Akt, and MAPK signaling pathway was significantly enriched among differentially expressed genes. AR reduced phosphorylation of PI3K, Akt, ERK, JNK, and p38, and markedly suppressed apoptosis in the spleen and bone marrow. Metabolomic profiling indicated that AR restored BDS-induced metabolic disturbances, while gut microbiota analysis showed that AR modulated microbial composition. Overall, AR produced more pronounced effects than RR. AR alleviated BDS in rats, potentially by inhibiting PI3K/Akt/MAPK-mediated apoptosis and restoring metabolic and microbial homeostasis. These findings support the traditional use of AR as a blood-tonifying herbal preparation.