Metabolomics study of Danggui Buxue Tang on treatment of type 2 diabetes mice using UHPLC-Q-TOF-MS / 中国中药杂志

In this paper, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS)-based metabolomics approach was used to explore the mechanism of Danggui Buxue Tang(DBT) in treating type 2 diabetes mellitus(T2 DM). T2 DM mice model was induced by high-sugar and high-fat fodder and streptozotocin(STZ). The routine indexes such as body weight, blood glucose, plasma insulin, IL-6 and related organ indexes were determined. The UHPLC-Q-TOF-MS technique was used to analyze the metabolism profile of serum samples between the control group and model group, and multiple statistical analysis methods including principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used to screen and identify biomarkers. Metabolic profiling revealed 16 metabolites as the most potential biomarkers distinguishing mice in model group from those in control group. The metabolomics pathway analysis(MetPA) was used to investigate the underlying metabolic pathways. Seven major metabolic pathways such the valine, leucine and isoleucine biosynthesis, glycerophospholipid metabolism, primary bile acid biosynthesis, taurine and hypotaurine metabolism, phenylalanine metabolism, fatty acid metabolism and biosynthesis of unsaturated fatty acid. Eleven metabolites such as taurocholic acid and palmitic acid were down-regulated in T2 DM mice, and five metabolites such as L-leucine and leukotriene E4 were up-regulated. Moreover, the sixteen biomar-kers of each administration group had a trend of returning to mice in control group. The significantly-altered metabolite levels indicated that DBT can improve the progression of type 2 diabetes by increasing insulin sensitivity, regulating sugar and lipid metabolism disorders, and relieving inflammation.

Effect of Yiqi Yangyin Zhuyu Recipe in Anti-inflammation Activities in Vitro on LPS-induced RAW 264.7 Cells / 中国实验方剂学杂志

Objective: To investigate the effect of Yiqi Yangyin Zhuyu recipe on expressions of inflammatory factor and transformation of classically activated macrophages(M1)/alternatively activated macrophages (M2) inflammatory phenotype in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Method: Methyl-thiazdyl-tetrazolium(MTT) reduction assay was used to detect the effect of different concentrations of Yiqi Yangyin Zhuyu recipe on the proliferation of the cells. The release of nitric oxide was detected by the Griess method. Enzyme linked immunosorbent assay(ELISA) was used to detect the release of M1/M2 inflammatory cytokines in cell supernatant. The expressions of the pro-inflammatory factor genes of M1-macrophages and the anti-inflammatory factor genes of M2-macrophages were detected by Real-time PCR. The protein expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1,nitric oxide synthase(iNOS) were detected by Western blot. Result: Results of MTT showed that Yiqi Yangyin Zhuyu recipe with the concentration of 2.0 g·L-1 and below had no effect on the cell proliferation. Results of Griess indicated that compared with blank group, the release of nitric oxide of LPS-induced group was increased (PPPPPPPα,IL-6,IL-1β,iNOS were up-regulated (Pα,IL-6,IL-1β,iNOS were down-regulated in Yiqi Yangyin Zhuyu recipe group, especially at the concentration at 2.0 g·L-1 (PConclusion: Yiqi Yangyin Zhuyu recipe could effectively inhibit the inflammatory reaction induced by LPS. The anti-inflammatory mechanism of Yiqi Yangyin Zhuyu recipe may be related to inhibition of macrophages to M1 phenotype polarization, so as to play the role of regulating immune and reducing the release of inflammatory cytokines, like NO,TNF-α,IL-6,IL-1β.

Protective Effect of Danggui Buxue Tang on Impaired Functional Activity of Endothelial Cells Exposed to Low Flow Shear Stress / 中国实验方剂学杂志

Objective: To investigate the protective effect of Danggui Buxue Tang(DGBX)with Angelicae Sinensis Radix(AS) and Astragali Radix(AR)at different radios on impaired functional activity of endothelial cells(ECs)exposed to low-fluid shear stress (FSS). Method: Low FSS was loaded by a parallel plate flow chamber,and ECs were divided into normal FSS group,low FSS group(each preincubated with M199 medium for 2 h),simvastatin group(preincubated with 0.1 μmol·L-1 simvastatin for 2 h),and 3 DGBX groups(preincubated with 3 g·L-1 AS and AR at 1:1,1:3,1:5 for 2 h,respectively). Then,the normal group was exposed to 1.2 Pa FSS,while the rest groups were all exposed to low FSS. At time points of 30,60, 360 min,the proliferation was detected by methyl thiazoly tetrazolium(MTT),the secretion of nitric oxide(NO) was detected by nitrase reduction test,and the mRNA and protein expressions of endothelial nitric oxide synthase(eNOS) were detected by Real-time fluorescence guantitative polymerase chain reaction(Real-time PCR) and Western blot,respectively. Result: Compared with the normal group, the secretion of NO and the expression of eNOS in ECs were both increased significantly at 60 min (PConclusion: DGBX could protect the functional activity of ECs exposed to low FSS.

Research Progress on Pharmacodynamic Material Basis and Quality Control of Danggui Buxue Tang / 中国实验方剂学杂志

Danggui Buxue Tang(DBT) is a famous classical formula of traditional Chinese medicine(TCM) for invigorating Qi and activating blood circulation.It is composed of Angelicae Sinensis Radix and Astragali Radix.It has the effect of enriching blood,invigorating Qi and regulating immunity,and has a high application value in clinical treatment.Because of its definite curative effect and simple compatibility,it has become a hot spot in the research field of TCM.This paper summarized and analyzed the research status of DBT from the pharmacological action,pharmacodynamic material basis,pharmacokinetics,quality control,compatibility and decoction process,so as to provide reference for the development of DBT and formulating its quality standard.

A novel approach based on metabolomics coupled with network pharmacology to explain the effect mechanisms of Danggui Buxue Tang in anaemia / 中国天然药物

Danggui Buxue Tang (DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague-Dawley(SD) rats were randomly divided into 3 groups including control (NC, Saline), the DBT (at a dose of 8.10 g), and blood deficiency(BD) (Cyclophosphamide (APH)-andCyclophosphamide(CTX)-induced anaemia). A metabolomics approach using Liquid Chromatography-Quadrupole-Time-of-Flight/Mass Spectrometry (LC/Q-TOFMS) was developed to perform the plasma metabolic profiling analysis and differential metaboliteswerescreened according to the multivariate statistical analysiscomparing the NC and BD groups, andthe hub metabolites were outliers with high scores of the centrality indices. Anaemia disease-related protein target and compound of DBT databases were constructed. The TCMSP, ChemMapper and STITCH databases were used to predict the protein targets of DBT. Using the Cytoscape 3.2.1 to establish a phytochemical component-target protein interaction network and establish a component, protein and hub metabolite protein-protein interaction (PPI) network and merging the three PPI networks basing on BisoGenet. The gene enrichment analysis was used to analyse the relationship between proteins based on the relevant genetic similarity by ClueGO. The results shown DBT effectively treated anaemia in vivo. 11 metabolic pathways are involved in the therapeutic effect of DBT in vivo; S-adenosyl-l-methionine, glycine, l-cysteine, arachidonic acid (AA) and phosphatidylcholine(PC) were screened as hub metabolites in APH-and CTX-induced anaemia. A total of 288 targets were identified as major candidates for anaemia progression. The gene-set enrichment analysis revealed that the targets are involved in iron ion binding, haemopoiesis, reactive oxygen species production, inflammation and apoptosis. The results also showed that these targets were associated with iron ion binding, haemopoiesis, ROS production, apoptosis, inflammation and related signalling pathways. DBT can promote iron ion binding and haemopoiesis activities, restrain inflammation, production of reactive oxygen, block apoptosis, and contribute significantly to the DBT treat anaemia.

Effect of Danggui Buxue Tang on immunological reconstitution of mice following bone marrow transplantation / 中华微生物学和免疫学杂志

Objective To study the effect of Danggui Buxue Tang (DBT) on immunological reconstitution of bone marrow transplantation (BMT) in mice. Methods BALB/c mice were irradiated by 137Cs γ once 8.5 Gy, then the mice were engrafted with bone marrow cells (107 cells/mouse) within 4 hours lethal irradiation. And the mice were fed by DBT every day for 15 days. Flow cytometry technique combined with immunological methods were performed to evaluate immunological reconstitution of BMT mice in 30 and 60 days pest-transplantation. Peripheral blood RBC and WBC were counted, and nucleated cells were assayed in recipient bone marrow. Lymphocyte numbers in thymus and periphery were counted and subpopulations of the two origins were observed respectively. Lymphocyte proliferation induced by Con A and LPS, plaque-forming cell (PFC), delayed type hypersensitivity (DTH) were also examined in 30,60 days in post-transplantation respectively. Results Compared with BMT mice, BMT mice treated with a certain dose of DBT could increase the number of peripheral blood RBC and WBC in the recipients, and also could increase that of nucleated cells significantly. BMT mice treated with DBT could increase lymphocyte numbers in thymus and periphery, and improve thymocyte subpopulations, resulting in enhancement in immune function. Conclusion DBT can enhance the immunological reconstitution of BMT mice.

Influence of Blood Replenishing Angelica Decoction on Enhanced Endothelial Permeability Induced by Histamine / 中草药

Blood Replenishing Angelica Decoction [Danggui Buxue Tang (DGBXT)]is a wellknown TCM prescription composed of Radix Angelicae Sinensis and Radix Astragali with the actions ofinvigorating "Qi" and enriching blood. Its action to curtail endothelial permeability induced by histaminewas studied. Endothelial cells isolated from the aorta of neonatal calf were cultured on polycarbonate mi-croporus filter membrane to develop a confluent endothelial monolayer. After purfused with either plainHank's balanced salt solution or that containing 5 g/L albumin, the monolayer was treated with 10-4 mol/L histamine for 30 min either with or without preincubation for 60 min with 10-4 g/mL of DGBXT. Fluidfiltration coefficient (Kf), filtration volume (Jv) and osmotic reflective coefficient (σ) of protein were thenmeasured. The findings showed that DGBXT could curtail the lowering of Kf and Jv and elevation of σ in-duced by histamine, indicating that DGBXT could inhibit the action of histamine on endothelial permeabili-ty, but its mechanism of action needs further study.

Summary of the experiment study on Angelica Decoction for Replenishing Blood / 中成药

Having referred the Chinese academic publications on TCM during the last decade ,we summarized the researches in Angelica Decoction (AD), which included immunoregulatory, circulatory system, the basis of decoction composition, anti-oxidation, liver injury and compaibility. AD can improve the immunity action, meliorate abnormal circulation, and also has the effects of anti-oxidation and protection of liver injury. Futher study on AD should be focused on its mechanism and material basis in the future.