Hepatic metabolomics combined with network pharmacology to reveal the correlation between the anti-depression effect and nourishing blood effect of Angelicae Sinensis Radix / 中国天然药物

Angelicae Sinensis Radix (AS) is reproted to exert anti-depression effect (ADE) and nourishing blood effect (NBE) in a rat model of depression. The correlation between the two therapeutic effects and its underlying mechanisms deserves further study. The current study is designed to explore the underlying mechanisms of correlation between the ADE and NBE of AS based on hepatic metabonomics, network pharmacology and molecular docking. According to metabolomics analysis, 30 metabolites involved in 11 metabolic pathways were identified as the potential metabolites for depression. Furthermore, principal component analysis and correlation analysis showed that glutathione, sphinganine, and ornithine were related to pharmacodynamics indicators including behavioral indicators and hematological indicators, indicating that metabolic pathways such as sphingolipid metabolism were involved in the ADE and NBE of AS. Then, a target-pathway network of depression and blood deficiency syndrome was constructed by network pharmacology analysis, where a total of 107 pathways were collected. Moreover, 37 active components obtained from Ultra Performance Liquid Chromatography-Triple-Time of Flight Mass Spectrometer (UPLC-Triple-TOF/MS) in AS extract that passed the filtering criteria were used for network pharmacology, where 46 targets were associated with the ADE and NBE of AS. Pathway enrichment analysis further indicated the involvement of sphingolipid metabolism in the ADE and NBE of AS. Molecular docking analysis indciated that E-ligustilide in AS extract exhibited strong binding activity with target proteins (PIK3CA and PIK3CD) in sphingolipid metabolism. Further analysis by Western blot verified that AS regulated the expression of PIK3CA and PIK3CD on sphingolipid metabolism. Our results demonstrated that sphingolipid metabolic pathway was the core mechanism of the correlation between the ADE and NBE of AS.

The mechanism of methotrexate in the treatment of rheumatoid arthritis using untargeted metabolomics / 中华风湿病学杂志

Objective:To investigate the mechanism of methotrexate in the treatment of rheumatoid arthritis (RA) by constructing a rat model of collagen-induced arthritis (CIA) and using non-targeted metabolomics.Methods:Enzyme-linked immunosorbent assay (ELISA) was used to determine the contents of TNF-α, IL-1β, IL-6, IL-4 and IL-10 in serum. HE staining and Masson staining were used to observe the histological changes of joints in each group. Non-targeted gas chromatography-mass spectrometry metabolomics technique was used to screen the expression profiles of differential metabolites in serum and cluster analysis and KEGG enrichment analysis were performed to screen the differential metabolic pathways, and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the key enzymes in the differential metabolic pathways. All experimental data conforming to the normal distribution were compared between groups using one-way ANOVA, and P<0.05 was considered statistically significant. Results:MTX significantly improved the joint inflammatory response and arthritis score and increased the body weight of CIA rats. The results of HE and Masson staining showed that MTX could ameliorate the erosion of articular cartilage by synovial tissue in CIA rats. ELISA results showed that MTX significantly decreased the contents of TNF-α [(191.2±17.4)pg/ml, F=40.31, P<0.001], IL-1β[(28.4±1.2)pg/ml, F=10.11, P=0.012] and IL-6[(118.7±1.4)pg/ml, F=829.40, P<0.001] in the serum and increased the contents of IL-4 [(49.3±3.3)pg/ml, F=33.44, P<0.001] and IL-10 [(30.2±0.7)pg/ml, F=33.44, P<0.001] in the serum of CIA rats. Non-targeted metabolomics technique showed MTX had an effect on metabolites such as phosphocholine, palmitic acid, oleic acid, and choline in the serum of CIA rats. KEGG pathway enrichment analysis showed that MTX had an effect on glycerophospholipid metabolism( P<0.01)and sphingolipid metabolism( P<0.05)in CIA rats. qRT-PCR results showed that MTX could down-regulate the expression of the key enzymes such as Plb1 [(1.00±0.49), F=8.23, P=0.019], Gpcpd1[(1.10±0.09), F=8.19, P=0.019], Chka [(1.33±0.19), F=33.00, P<0.001], Chkb [(2.07±1.21), F=8.20, P=0.019]and Phospho1 [(1.07±0.14), F=13.58, P=0.006]in the glycerophospholipid metabolic pathway in the synovial membrane of CIA rats, and can also down-regulate the expression of the key enzymes Kdsr [(1.24±0.32), F=13.85, P=0.006], Plpp1 [(1.61±0.32), F=11.95, P=0.003) and Degs1 [(1.21±0.15, F=46.55, P<0.001]in the sphingolipid metabolic pathway. Conclusion:The biological mechanism of MTX in the treatment of rheumatoid arthritis may be related to the down-regulation of glycerophospholipid metabolism and sphingolipid metabolism pathway metabolic levels in the body.

Regulatory effect of Flos Abelmoschus manihot in mice with inflammatory bowel disease based on gut microbiota sequencing and untargeted lipidomics / 药学学报

In this study, the ameliorative effects of Flos Abelmoschus manihot on mice with chronic inflammatory bowel disease (IBD) were investigated and its effects on the structure of the intestinal flora as well as the lipid profile in feces of IBD mice were analyzed. All animal welfare and experimental procedures followed the regulations of the Animal Ethics Committee of Nanjing University of Chinese medicine. A mouse model with chronic IBD induced by dextran sulfate sodium (DSS) was used to evaluate changes in body weight, disease activity index (DAI), colonic histopathological damage as well as gene expression levels of inflammatory factors in the colon. Fecal samples from mice in each group were collected and subjected to Illumina high-throughput sequencing to detect the abundance of intestinal flora; samples were analyzed by UHPLC-Q-Exactive® HF Quadrupole-Orbitrap® of untargeted lipidomics, which detects lipid content in feces. Administration of Flos Abelmoschus manihot could significantly restore the body weight and ameliorate colonic histopathological damage in IBD mice. Sequencing of the gut microbiota revealed that the species diversity and richness of the gut microbiota in IBD mice were decreased, with a significant increase in the abundance of Verrucomicrobia and a significant decrease in the abundance of Bacteroidetes; Flos Abelmoschus manihot significantly increased the richness and diversity of intestinal microbiota in IBD mice, increased the number of taxa species at each level, and restored the abundance of bacteria in the phylum Bacteroidetes. Analysis of fecal lipid profiles identified the most significant changes in sphingolipid and glycerophospholipid metabolic pathways in IBD mice, with Flos Abelmoschus manihot inhibiting ceramide and sphingomyelin synthesis in sphingolipid metabolism. In summary, Flos Abelmoschus manihot can effectively improve the disease condition of mice with chronic IBD, and it has the effect of regulating intestinal flora homeostasis and lipid metabolism, but the related mechanism between the two still needs to be deeply explored.

Mechanism of emodin in relieving neuropathic pain by regulating serum metabolism / 中国中药杂志

The present study investigated the effect of emodin on the serum metabolite profiles in the chronic constriction injury(CCI) model by non-target metabolomics and explored its analgesic mechanism. Twenty-four Sprague Dawley(SD) rats were randomly divided into a sham group(S), a CCI group(C), and an emodin group(E). The rats in the emodin group were taken emodin via gavage once a day for fifteen days(50 mg·kg~(-1)) on the first day after the CCI surgery. Mechanical withdrawal threshold(MWT) and thermal withdrawal threshold(TWL) in each group were performed before the CCI surgery and 3,7, 11, and 15 days after surgery. After 15 days, blood samples were collected from the abdominal aorta. The differential metabolites were screened out by non-target metabolomics and analyzed with Kyoto Encyclopedia of Genes and Genomes(KEGG) and ingenuity pathway analysis(IPA). From the third day after CCI surgery, the MWT and TWL values were reduced significantly in both CCI group and emodin group, compared with the sham group(P<0.01). At 15 days post-surgery, the MWT and TWL values in emodin group increased significantly compared with the CCI group(P<0.05). As revealed by non-target metabolomics, 72 differential serum metabolites were screened out from the C-S comparison, including 41 up-regulated and 31 down-regulated ones, while 26 differential serum metabolites from E-C comparison, including 10 up-regulated and 16 down-regulated ones. KEGG analysis showed that the differential metabolites in E-C comparison were enriched in the signaling pathways, such as sphingolipid metabolism, arginine biosynthesis, glycerophospholipid metabolism, and tryptophan metabolism. IPA showed that the differential metabolites were mainly involved in the lipid metabolism-molecular transport-small molecule biochemistry network. In conclusion, emodin can exert an analgesic role via regulating sphingolipid metabolism and arginine biosynthesis.