Integration of gut microbiota and metabolomics for the hematopoiesis of Siwu paste on anemia rats.

Background: To investigate the regulation mechanism of hematopoiesis of Siwu paste (SWP) in anemia rats, which is a classic Chinese prescription used for nourishing blood or blood deficiency over 1000 years. Methods: Blood cell and biochemical analysis were used to evaluate the hematopoietic function of SWP in anemia rats. The intestinal microbial composition was analyzed with 16S rRNA gene sequencing, and the metabolites were profiled using UPLC-TripleTOF system nontargeting metabolomics. Results: SWP can improve the levels of red blood cells, hemoglobin, platelet, hematocrit value, white blood cells, lymphocyte, EPO, TPO, and GM-CSF in anemia rats, and significantly change the microbial community and its metabolites. The correlation analysis of intestinal microbiota-hematopoietic efficacy shows that 13 kinds of different intestinal flora were related to hematopoietic efficacy, in which Prevotella_1, Prevotella_9, Lactobacillus, and norank_f__Muribaculaceae were significantly positively correlated with hematopoiesis, nine kinds of intestinal flora are negatively correlated with hematopoietic effect. Compared with anemia rats, 218 potential metabolic biomarkers and 36 metabolites with significant differences were identified in the SWP treatment group, and the key metabolites were mainly amino acids and lipids. An in-depth analysis of metabolic pathways showed that SWP mainly affected 7 metabolic pathways, including aminobenzoic acid degradation and tryptophan metabolism. Conclusion: The study provides novel insights into the regulation of hematopoiesis of SWP in anemia rats that were correlated with gut microbiota and the metabolites, which through the restoration of the firmicutes/bacteroidetes ratio.

Integrated Network Pharmacology and Metabolomics Analysis to Reveal the Potential Mechanism of Siwu Paste on Aplastic Anemia Induced by Chemotherapy Drugs.

Purpose: This study aimed to reveal the multicomponent synergy mechanisms of SWP based on network pharmacology and metabolomics for exploring the relationships of active ingredients, biological targets, and crucial metabolic pathways. Materials: Network pharmacology, including TRRUST, GO, and KEGG, enrichment was used to discover the active ingredients and potential regulation mechanisms of SWP. LC-MS and multivariate data analysis method were further applied to analyze serum metabolomics profiling for discovering the potential metabolic mechanisms of SWP on AA induced by Cyclophosphamide (CTX) and 1-Acetyl-2-phenylhydrazine (APH). Results: A total of 27 important bioactive ingredients meeting the ADME (absorption, distribution, metabolism, and excretion) screening criteria from SWP were selected. Interaction networks were constructed and validated based on the 10 associated ingredients with the relevant targets. A total of 125 biomarkers were found by Metabolomics approach, which associated with the development of AA, mainly involved in amino acid metabolism and lipid metabolism. While SWP can reverse the above 12 metabolites changed by AA. Network analysis revealed the synergistic effects of SWP through the 43 crucial pathways, including Sphingolipid signaling pathway, Sphingolipid metabolism, Arginine and proline metabolism, VEGF signaling pathway, Estrogen signaling pathway. Conclusion: The study suggested that SWP is a useful alternative for the treatment of AA induced by CTX + APH. Its potential mechanisms are to improve hematopoietic microenvironment and promote bone marrow hematopoiesis therapies.

Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism / 中国实验方剂学杂志

Objective:To explore the regulatory effect of Siwu paste on the bone marrow hematopoietic function of aplastic anemia （AA） model rats. Method:SD rats were randomly divided into normal control group， model group， positive drug （Fufang E'jiao Jiang 10.8 g·kg^-1） group， high-dose Siwu paste （22.68 g·kg^-1） group and low-dose Siwu paste （5.67 g·kg^-1） group. Acetophenazine （APH） combined with cyclophosphamide （CTX） injection was used to establish the aplastic anemia rat model. The administration groups were given the corresponding drugs （<italic>ig</italic>） for 15 consecutive days. The levels of white blood cells （WBC）， red blood cells （RBC）， hemoglobin （HGB）， hematocrit （HCT） and platelets （PLT） in peripheral blood cells of rats were detected， thymus and spleen indexes were calculated and compared. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-3 （IL-3） and interleukin-6 （IL-6） in rat serum. The pathological changes of bone marrow were observed by hematoxylin eosin （HE） staining. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） methods were used to detect Toll receptor 4 （TLR4） and nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） protein and gene expression in rat femoral bone marrow cells. Result:Compared with the normal control group， the WBC， RBC， HGB， HCT and PLT levels of the model group were significantly reduced， the thymus index was significantly decreased， the spleen index was significantly increased， the serum IL-3 level was significantly decreased， and the IL-6 level was significantly increased. The number of neutrophils and megakaryocytes in the femoral bone marrow was reduced， and the medullary cavity was filled with edema fibrofatty tissue. The expressions of TLR4 and NF-<italic>κ</italic>B protein and mRNA in bone marrow cells were significantly increased （<italic>P</italic><0.01）. Compared with the model group， the levels of WBC， RBC， HCT and PLT in peripheral blood cells of rats in the high-dose Siwu paste group increased， the thymus index increased， the spleen index decreased， the IL-3 level was significantly increased， the IL-6 level was significantly decreased， the pathological morphology of femoral bone marrow was slightly improved， and the expressions of TLR4 and NF-<italic>κ</italic>B protein and mRNA in bone marrow cells decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Siwu paste may improve the bone marrow hematopoietic function of rats with aplastic anemia by regulating the expression of the bone marrow inflammation signal pathway TLR4/NF-<italic>κ</italic>B.