Sanhuang xiexin decoction synergizes insulin/PI3K-Akt/FoxO signaling pathway to inhibit hepatic glucose production and alleviate T2DM.

ETHNOPHARMACOLOGICAL RELEVANCE: Sanhuang Xiexin Decoction (SHXXD) is a classic prescription for the treatment of diabetes. Excessive hepatic glucose production (HGP) is a major determinant of the occurrence and development of diabetes. Inhibition of HGP can significantly improve type 2 diabetes mellitus (T2DM). AIM OF THE STUDY: To investigate the mechanism by which SHXXD inhibits HGP. MATERIALS AND METHODS: First, a mouse model of T2DM was established through high-fat diet (HFD) feeding combined with streptozotocin (STZ) injection to determine the pharmacodynamic effect of SHXXD in T2DM mice. Then, the possible pathways induced by SHXXD in the treatment of T2DM were predicted by network pharmacology combined with transcriptomics (including target prediction, network analysis and enrichment analysis). Finally, the specific mechanism of SHXXD was elucidated by in vitro experiments. RESULTS: In vivo experiments showed that SHXXD reduced fasting blood glucose and alleviated weight loss in T2DM mice. Improved glucose clearance rates and insulin sensitivity improve dyslipidemia, liver tissue structural abnormalities and inflammatory cell infiltration as well as increase glycogen storage in T2DM mice. The results of network pharmacology and transcriptome analysis showed that SHXXD contained 378 compounds and 2625 targets. In total, 292 intersection targets were identified between the differentially expressed genes (DEGs) of the liver tissue insulin resistance (IR) related dataset GSE23343. KEGG enrichment analysis showed that the insulin/PI3K-Akt/FoxO signaling pathway may be related to SHXXD-mediated improvements in T2DM. In vitro experimental results showed that SHXXD increased glucose consumption by HepG2-IR cells and improved their insulin sensitivity. RT‒qPCR and Western blotting results showed that SHXXD inhibited hepatic gluconeogenesis through the insulin/PI3K-Akt/FoxO signaling pathway by promoting IGFIR, PIK3R1 and AKT2 expression and subsequently inhibiting PEPCK and FBP1 expression via phosphorylation of Foxo1. In addition, PI3K/Akt deactivated p-GSK3ß through phosphorylation, thereby promoting GS expression and increasing glycogen synthesis. CONCLUSIONS: SHXXD can target the liver to cooperate with the insulin/PI3K-Akt/FoxO signaling pathway to inhibit HGP to alleviate T2DM.

Mechanisms of Danggui Buxue Tang on Hematopoiesis via Multiple Targets and Multiple Components: Metabonomics Combined with Database Mining Technology.

This study aimed to explore the mechanism of action of Danggui Buxue Tang (DBT) with its multiple components and targets in the synergistic regulation of hematopoiesis. Mouse models of hematopoiesis were established using antibiotics. Metabolomics was used to detect body metabolites and enriched pathways. The active ingredients, targets, and pathways of DBT were analyzed using system pharmacology. The results of metabolomics and system pharmacology were integrated to identify the key pathways and targets. A total of 515 metabolites were identified using metabolomics. After the action of antibiotics, 49 metabolites were markedly changed: 23 were increased, 26 were decreased, and 11 were significantly reversed after DBT administration. Pathway enrichment analysis showed that these 11 metabolites were related to bile secretion, cofactor biosynthesis, and fatty acid biosynthesis. The results of the pharmacological analysis showed that 616 targets were related to DBT-induced anemia, which were mainly enriched in biological processes, such as bile secretion, biosynthesis of cofactors, and cholesterol metabolism. Combined with the results of metabolomics and system pharmacology, we found that bile acid metabolism and biotin synthesis were the key pathways for DBT. Forty-two targets of DBT were related to these two metabolic pathways. PPI analysis revealed that the top 10 targets were CYP3A4, ABCG2, and UGT1A8. Twenty-one components interacted with these 10 targets. In one case, a target corresponds to multiple components, and a component corresponds to multiple targets. DBT acts on multiple targets of ABCG2, UGT1A8, and CYP3A4 through multiple components, affecting the biosynthesis of cofactors and bile secretion pathways to regulate hematopoiesis.

Gut microbiota affects the efficacy of Danggui Buxue Tang by affecting plasma concentration of active ingredients.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui Buxue Tang (DBT) is a traditional Chinese medicine, which has the function of supporting Qi and enriching blood. Antibiotics can cause Gut microbiota disorder and affect efficacy of DBT. AIM OF THE STUDY: Explore the manner in which Gut microbiota affects the efficacy of Danggui Buxue Tang. MATERIALS AND METHODS: In this study, antibiotics were used to destroy gut microbiota. The changes of DBT efficacy were detected to verify the effect of gut microbiota on DBT efficacy. The changes of gut microbiota was detected using 16S rRNA sequencing, and UPLC-MS/MS was used to analyze the plasma concentration of active ingredients. Correlation analysis was used to establish the relationship between gut microbiota, blood components and drug efficacy, and to explore the role of gut microbiota in the efficacy of DBT. RESULTS: The results showed that the efficacy in the DBT group was significantly improved compared with the control group (p<0.05). Compared with DBT group, the efficacy in antibiotic DBT treatment (ABXDBT) group was significantly reduced, 194 plasma metabolites and 18 DBT blood components were significantly altered in ABXDBT group, and 11 DBT blood components such as caffeic acid and formononetin were significantly decreased. Correlation analysis showed that 6 DBT blood components were related with the decrease of efficacy. Network pharmacology analysis showed that the above 6 DBT blood components participated in the hematopoietic regulation through PI3K-Akt and HIF-1 signaling pathways. Correlation analysis showed that Bacteroides and other intestinal bacteria were related to the absorption of DBT active ingredients. The drug metabolic pathway of gut microbiota was significantly decreased after antibiotic treatment (p = 0.033). CONCLUSIONS: Gut microbiota such as Bacteroides affects the efficacy of DBT by affecting the metabolism and absorption of DBT active ingredients such as caffeic acid and formononetin.

Danggui Buxue Tang restores antibiotic-induced metabolic disorders by remodeling the gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui Buxue Tang (DBT) has been used to promote hematopoiesis and relieve myelosuppression in China. Antibiotics can cause myelosuppression through gut microbiota disorders. AIM OF THE STUDY: This study aims to explore the way of DBT to alleviate the metabolic disorder caused by antibiotics. MATERIALS AND METHODS: In this study, 16S rRNA sequencing was used to detect the change of gut microbiota, metabolomics to analyze the change of metabolites. Correlation analysis was used to establishment the correlation between gut microbiota and metabolites. PICRUST 2 was used to predict the function of gut microbiota. RESULTS: Results showed that eighty-two genera of gut microbiota were affected by antibiotic, while twelve were significantly restored after DBT. Seventy-four potential metabolites were significantly different from the antibiotics and DBT. We found significant recovery by the Bacteroides and Rikenellaceae RC9 after DBT. The metabolic pathways influenced by the antibiotic treatment included primary and secondary bile biosynthesis, etc. The metabolic pathways that could be restored after DBT included the primary and secondary bile acid biosynthesis pathway, etc. Through correlation analysis, we found a correlation between the Bacteroides, Rikenellaceae_RC9_gut_group and other potential differential metabolisms such as those of taurodeoxycholic acid, N-phenylacetyl glycine, etc. The functional prediction showed that the biosynthesis of primary bile acid, secondary bile acid was significantly affected. CONCLUSIONS: DBT can restore the gut and reverse the metabolic disorder caused by antibiotics through Bacteroides, and it provides a new medical idea regarding the gut microbiota balance.

Metabolomic analysis of serum reveals the potential effective ingredients and pathways of Danggui Buxue Tang in promoting erythropoiesis.

Danggui Buxue Tang has been used for menopausal women in China for more than 800 years. However, the potential effective ingredients and pathways require further investigation. The main objective of this work was to explore the potential effective ingredients and pathways. The optimal administration time was optimized by detecting the changes of reticulocytes in peripheral blood. Drug-containing serum (DCS) was taken every 30 min after last administration. Because of the different concentration of effective ingredients absorbed into blood at different time, the pharmacodynamic effect is different. Therefore, bone marrow stromal cells as a member of hematopoietic microenvironment were used to evaluate the pharmacodynamics of DCS. Metabolomics was used to detect changes of metabolites (DBT and endogenous metabolites). The correlation of the metabolites and pharmacodynamics was used to identify the metabolites associated with erythropoiesis. After 14 days, the number of reticulocytes in peripheral blood, erythroid-related cells and erythroid progenitor cells in bone marrow in the DBT group were significantly increased. In vitro experiments showed that DCS at different time had different proliferation effects on BMSCs. Metabolomic analysis showed that the concentration of metabolites in DCS at different time was significantly different. The correlation analysis identified 7 DBT metabolites and 15 endogenous metabolites related to erythropoiesis. 15 endogenous metabolites were finally connected to different pathways. Glutamate is a node molecule. 7 potential effective ingredients of DBT were found. DBT promoted erythropoiesis via promoting the metabolism of glutamate and further affect other pathways.

Danggui Buxue Tang promotes the adhesion and migration of bone marrow stromal cells via the focal adhesion pathway in vitro.

ETHNOPHARMACOLOGY RELEVANCE: Danggui Buxue Tang has been used in China to treat clinical anemia for more than 800 years. However, there is no scientific report on its effect on bone marrow stromal cells. AIM OF THE STUDY: Here, we aimed to explore the effect of Danggui Buxue Tang on bone marrow stromal cell adhesion and migration. MATERIALS AND METHODS: Bone marrow stromal cells were used as a model to evaluate the effect of Danggui Buxue Tang on the adhesion and migration of bone marrow stromal cells. RNA-sequencing, quantitative polymerase chain reaction, and western blotting were used to detect and confirm the expression of genes related to the focal adhesion pathway before and after drug delivery. RESULTS: Danggui Buxue Tang significantly increased the number of bone marrow stromal cells. After 12 days of 16 mg/mL Danggui Buxue Tang treatment, bone marrow stromal cells were significantly increased (by 0.527 ±â€¯0.008 fold; p < 0.001) as compared to the control group (0.180 ±â€¯0.019). The effect was not due to enhanced cell proliferation, as there was no difference in the cell cycle (p > 0.05). The adhesion area of a single cell was doubled by Danggui Buxue Tang treatment (p < 0.001), and the time required for cell adhesion to a Petri dish was shortened. Thus, Danggui Buxue Tang increases the number of bone marrow stromal cells by promoting adhesion. Danggui Buxue Tang also significantly promoted bone marrow stromal cell migration (p <  0.001). Transcript analysis revealed that the focal adhesion and PI3K-Akt signaling pathways were activated. Expression analysis confirmed that the gene and protein expression of focal adhesion-related factors were upregulated. CONCLUSION: Danggui Buxue Tangaffects bone marrow stromal cell adhesion and migration by enhancing the focal adhesion pathway in vitro, and bone marrow stromal cells are a target of DBT-regulated hematopoiesis, and the active ingredients of DBT involved in the effects require further investigation.