Flos Magnoliae Research Articles

Comparative metabolism of aschantin in human and animal hepatocytes.

Aschantin, a tetrahydrofurofuran lignan with a 1,3-benzodioxole group derived from Flos Magnoliae, exhibits antioxidant, anti-inflammatory, cytotoxic, and antimicrobial activities. This study compared the metabolic profiles of aschantin in human, dog, mouse, and rat hepatocytes using liquid chromatography-high-resolution mass spectrometry. The hepatic extraction ratio of aschantin among the four species was 0.46-0.77, suggesting that it undergoes a moderate-to-extensive degree of hepatic metabolism. Hepatocyte incubation of aschantin produced 4 phase 1 metabolites, including aschantin catechol (M1), O-desmethylaschantin (M2 and M3), and hydroxyaschantin (M4), and 14 phase 2 metabolites, including O-methyl-M1 (M5 and M6) via catechol O-methyltransferase (COMT), six glucuronides of M1, M2, M3, M5, and M6, and six sulfates of M1, M2, M3, M5, and M6. Enzyme kinetic studies using aschantin revealed that the production of M1, a major metabolite, via O-demethylenation is catalyzed by cytochrome 2C8 (CYP2C8), CYP2C9, CYP2C19, CYP3A4, and CYP3A5 enzymes; the formation of M2 (O-desmethylaschantin) is catalyzed by CYP2C9 and CYP2C19; and the formation of M4 is catalyzed by CYP3A4 enzyme. Two glutathione (GSH) conjugates of M1 were identified after incubation of aschantin with human and animal liver microsomes in the presence of nicotinamide adenine dinucleotide phosphate and GSH, but they were not detected in the hepatocytes of all species. In conclusion, aschantin is extensively metabolized, producing 18 metabolites in human and animal hepatocytes catalyzed by CYP, COMT, UDP-glucuronosyltransferase, and sulfotransferase. These results can help in clarifying the involvement of metabolizing enzymes in the pharmacokinetics and drug interactions of aschantin and in elucidating GSH conjugation associated with the reactive intermediate formed from M1 (aschantin catechol).

Comparative metabolism of fargesin in human, dog, monkey, mouse, and rat hepatocytes.

Fargesin, a bioactive lignan derived from Flos Magnoliae, possesses anti-inflammatory, anti-oxidative, anti-melanogenic, and anti-apoptotic effects. This study compared the metabolic profiles of fargesin in human, dog, monkey, mouse, and rat hepatocytes using liquid chromatography-high resolution mass spectrometry. In addition, we investigated the human cytochrome P450 (CYP), UDP-glucuronosyltransferase (UGT), and sulfotransferase (SULT) enzymes responsible for fargesin metabolism. The hepatic extraction ratio of fargesin among the five species ranged from 0.59 to 0.78, suggesting that it undergoes a moderate-to-extensive degree of hepatic metabolism. During metabolism, fargesin generates three phase 1 metabolites, including fargesin catechol (M1) and O-desmethylfargesin (M2 and M3), and 11 phase 2 metabolites, including O-methyl-M1 (M4 and M5) via catechol O-methyltransferase (COMT), glucuronides of M1, M2, M4, and M5, and sulfates of M1-M5. The production of M1 from fargesin via O-demethylenation is catalyzed by CYP2C9, CYP3A4, CYP2C19, and CYP2C8 enzymes, whereas the formation of M2 and M3 (O-desmethylfargesin) is catalyzed by CYP2C9, CYP2B6, CYP2C19, CYP3A4, CYP1A2, and CYP2D6 enzymes. M4 is metabolized to M4 glucuronide by UGT1A3, UGT1A8, UGT1A10, UGT2B15, and UGT2B17 enzymes, whereas M4 sulfate is generated by multiple SULT enzymes. Fargesin is extensively metabolized in human hepatocytes by CYP, COMT, UGT, and SULT enzymes. These findings help to elucidate the pharmacokinetics and drug interactions of fargesin.

Effect of Traditional Chinese Medicine on Allergic Rhinitis in Children under Data Mining.

The data mining analysis of the medication rule and the curative effect of traditional Chinese medicine in treating allergic rhinitis in children was performed by using the association rule Apriori algorithm. The model of interest degree was introduced to improve the Apriori algorithm, and the performance difference of the algorithm before and after improvement was analyzed. Traditional Chinese medicine prescriptions for the treatment of allergic rhinitis in children were selected from the dictionary of Chinese medicine formulations. The frequency, frequent itemsets, and the improved Apriori algorithm of each prescription were analyzed comprehensively. The results showed that both the execution time of the improved Apriori algorithm and the number of mining association rules were signally lower. 102 Chinese herbal compounds were selected, in which the occurrence frequency of Flos magnoliae was the highest (67 times, 5.33%). The occurrence frequency of diaphoretic drugs was the highest (412 times, 32.78%) in drug types. The occurrence frequency of Yu Ping Feng powder was the highest (21 times, 20.59%) in the Chinese herbal compound. After the association rule analysis of the improved Apriori algorithm, Perilla frutescens, Saposhnikovia divaricata, ginseng, Notopterygium root, and Astragalus propinquus Schischkin were often mixed with liquorice, and Flos magnoliae were usually mixed with Fructus xanthii and black plum. Compared with the conditions before treatment, the sign scores of children with allergic rhinitis were remarkably decreased after treatment with traditional Chinese medicine compounds (P < 0.05). The mining performance of the Apriori algorithm was improved by introducing an interest-based model. The treatment of traditional Chinese medicine on allergic rhinitis in children was combined with children's physiological and pathological characteristics of children, which used mild medicines.

Pectin polysaccharide from Flos Magnoliae (Xin Yi, Magnolia biondii Pamp. flower buds): Hot-compressed water extraction, purification and partial structural characterization

In this study, three novel and natural polysaccharides (FMP-1, FMP-2, and FMP-3) were obtained from Flos Magnoliae (FM, Xinyi in Chinese) using an efficient and green technique (hot-compressed water) for extraction and purified by DEAE-52 and Sephadex G-150. Their physicochemical properties, morphological and structural characteristics were investigated using chemical methods and instrumental analysis including methylation, partial acid hydrolysis, STA, GC-MS, HPGPC, HPAEC, FT-IR and NMR (1D and 2D). Results showed that FMPs all contained arabinose, rhamnose, galactose, xylose, mannose and galacturonic acid. FMP-2, with the highest yield and average molecular weight of the three (278.69 kDa), had large amounts of RG-I, minor amounts of HG with the molar ratio of 83.5:10.4, and trace amounts of RG-II domains. Thermal analys is exhibited that FMP-2 has a favorable thermal stability and it also exhibited effective antioxidant activities in a dose-dependent manner, which could be used as a potential stabilizer, thickening agent and natural antioxidant. These investigations provide a scientific basis for further investigation of FM polysaccharides in the context of their potential application in functional foods and medicines.

Flos magnoliae constituent fargesin has an anti-allergic effect via ORAI1 channel inhibition.

Flos magnoliae (FM), the dry flower buds of Magnolia officinalis or its related species, is a traditional herbal medicine commonly used in Asia for symptomatic relief of and treating allergic rhinitis, headache, and sinusitis. Although several studies have reported the effects of FM on store-operated calcium entry (SOCE) via the ORAI1 channel, which is essential during intracellular calcium signaling cascade generation for T cell activation and mast cell degranulation, the effects of its isolated constituents on SOCE remain unidentified. Therefore, we investigated which of the five major constituents of 30% ethanoic FM (vanillic acid, tiliroside, eudesmin, magnolin, and fargesin) inhibit SOCE and their physiological effects on immune cells. The conventional whole-cell patch clamp results showed that fargesin, magnolin, and eudesmin significantly inhibited SOCE and thus human primary CD4+ T lymphocyte proliferation, as well as allergen-induced histamine release in mast cells. Among them, fargesin demonstrated the most potent inhibitory effects not only on ORAI1 (IC50 = 12.46 ± 1.300 μM) but also on T-cell proliferation (by 87.74% ± 1.835%) and mast cell degranulation (by 20.11% ± 5.366%) at 100 μM. Our findings suggest that fargesin can be a promising candidate for the development of therapeutic drugs to treat allergic diseases.

Preparation and quality evaluation of a volatile oil microemulsion from Flos magnoliae and Centipeda minima.

In order to improve the water solubility of the volatile oils extracted from Flos magnoliae (FM) and Centipeda minima (CM), they were prepared as a microemulsion (ME), which were then used in the development of an FM and CM volatile oil ME for the treatment of allergic rhinitis (AR). ME was prepared by phase inversion emulsification, and the prescription factors such as emulsifier, co-emulsifier, oil phase, Km, which represents the ratio of the mass of emulsifier to that of the co-emulsifier, and preparation factors such as temperature affecting the formation of the ME were selected according to the formation area of ME in a pseudo-ternary phase diagram. The quality of the ME was evaluated based on its appearance, particle size, Zeta potential and stability. The content of eucalyptol in ME was determined by gas chromatography-mass spectrometry (GC-MS). The cumulative permeability of the ME within 24 h was measured with a transdermal diffusion tester. The results revealed that the best formula for preparation of the ME was as follows: Castor oil polyoxyethylene ether (EL-40) was the emulsifier; the co-emulsifier was anhydrous ethanol; the Km was 2:1; the mixed phase of volatile oil and isopropyl myristate with mass ratio of 1:1 was used as oil phase; and the preparation temperature was 25°C. The content of eucalyptol in the ME was 2.57 mg/g, and the cumulative permeability of the ME in 24 h was significantly increased compared with that of the reference oil solution. The appearance of the ME was uniform, and the solution was transparent. In conclusion, compared with traditional preparations, FM and CM volatile oil ME is a novel, improved and more effective preparation for the treatment of AR.

Flos Magnoliae and Its Constituent Linoleic Acid suppress T Lymphocyte Activation via Store‐Operated Calcium Entry.

Intracellular calcium signaling is crucial for type 2 helper T cell and mast cell activation, which is essential for allergic inflammation. It is initiated by antigen‐mediated receptor stimulation that triggers store‐operated calcium entry (SOCE) via ORAI1 calcium channel. Flos Magnoliae (FM) is widely used to treat allergic diseases such as allergic rhinitis and asthma. Although many studies have reported that FM regulates intracellular calcium signaling, research on the exact type of calcium channel modulated by FM is scarce. Therefore, we hypothesized that the anti‐allergic effects of FM might result from ORAI1 inhibition in T cells. We investigated whether a 70% ethanolic extract of FM (FMEtOH) and its constituents inhibit ORAI1 channel activity and subsequent T cell activation. We performed conventional whole‐cell patch clamp studies in STIM1 and ORAI1‐overexpressing HEK293T cells (HEKORAI1). Intracellular calcium concentration was determined using Fura‐2 dye and cytokine production measurement in Jurkat T lymphocytes. FMEtOH (0.03 mg/mL) and its fractions, especially hexane fraction (FMHex, 0.01 mg/mL), significantly inhibited SOCE and IL‐2 cytokine production in Jurkat T lymphocytes. GC/MS analysis showed linoleic acid (LA) as the major component of FMHex. FMHex at 0.01 mg/mL (equivalent to 10 μM LA) inhibited not only SOCE but also IL‐2 production, as well as CD3/CD28 receptor co‐stimulation induced calcium signaling in Jurkat T lymphocytes. FMEtOH and LA suppressed CD4+ T lymphocyte activation, at least in part, by inhibiting ISOCE. Thus, ISOCE inhibition may be a potential strategy to inhibit immune responses in inflammation.Linoleic acid (LA) inhibits ISOCE and IL‐2 production to similar levels.Figure 1FMEtOH and its fractions inhibit store‐operated Ca2+ entry mediated current (ISOCE) in ORAI1 and STIM1 co‐overexpressing HEK293T cells (HEKORAI1).Figure 2

Pharmacology mechanism of Flos magnoliae and Centipeda minima for treating allergic rhinitis based on pharmacology network

Chinese herbs such as Flos magnoliae (FM) and Centipeda minima (CM) can be effective in treating allergic rhinitis (AR). However, there is little research on the therapeutic mechanism of these two drugs acting on AR at the same time. In order to systematically understand the mechanism of action of two drugs acting on AR at the same time, we searched various databases to obtain 31 components and 289 target proteins of FM, 25 components and 465 target proteins of CM. The interaction networks of FM, CM, and AR proteins were constructed by Cytoscape-v3.2.1 software. The core protein of two network intersections was obtained by using Venny 2.1.0. The R platform was used for the core target protein gene ontology (GO) comment analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis. Thirteen common targets and seven acting pathways were obtained. The results of animal experiments showed that FM and CM volatile oil could effectively improve the symptoms of AR by regulating the common targets. In summary, this study successfully explained the potential therapeutic mechanism of FM and CM in the treatment of AR. At the same time, it indicates that the two drugs can be compatible as a new application.

Flos Magnoliaeand its Constituent Linoleic Acid Suppress T Lymphocyte ActivationviaStore-Operated Calcium Entry

Intracellular calcium signaling is crucial for type2 helper T cell and mast cell activation, which is essential for allergic inflammation. It is initiated by antigen-mediated receptor stimulation that triggers store-operated calcium entry (SOCE) via ORAI1 calcium channel. Flos Magnoliae (FM) is widely used to treat allergic diseases such as allergic rhinitis and asthma. Although many studies have reported that FM regulates intracellular calcium signaling, research on the exact type of calcium channel modulated by FM is scarce. Therefore, we hypothesized that the anti-allergic effects of FM might result from ORAI1 inhibition in T cells. We investigated whether a 70% ethanolic extract of FM (FM and its constituents inhibit ORAI1 channel activity and subsequent T cell activation. We performed conventional whole-cell patch clamp studies in hSTIM1 and hORAI1-overexpressing HEK293T cells (HEK. Intracellular calcium concentration was determined using Fura-2 dye and cytokine production measurement in Jurkat T lymphocytes. FMEtOH (0.03 mg/mL) and its fractions, especially hexane fraction (FMHex, 0.01 mg/mL), significantly inhibited SOCE and IL-2 cytokine production in Jurkat T lymphocytes. GC/MS analysis showed linoleic acid (LA) as the major component of FMHex. FMHex at 0.01 mg/mL (equivalent to 10 M LA) inhibited not only SOCE but also IL-2 production, as well as CD3/CD28 receptor co-stimulation induced calcium signaling in Jurkat T lymphocytes. FMEtOH and LA suppressed CD4 T lymphocyte activation, at least in part, by inhibiting . Thus, inhibition may be a potential strategy to inhibit immune responses in inflammation.

Anti-Inflammatory Effects of Fargesin on Chemically Induced Inflammatory Bowel Disease in Mice.

Fargesin is a bioactive lignan from Flos Magnoliae, an herb widely used in the treatment of allergic rhinitis, sinusitis, and headache in Asia. We sought to investigate whether fargesin ameliorates experimental inflammatory bowel disease (IBD) in mice. Oral administration of fargesin significantly attenuated the symptoms of dextran sulfate sodium (DSS)-induced colitis in mice by decreasing the inflammatory infiltration and myeloperoxidase (MPO) activity, reducing tumor necrosis factor (TNF)-α secretion, and inhibiting nitric oxide (NO) production in colitis mice. The degradation of inhibitory κBα (IκBα), phosphorylation of p65, and mRNA expression of nuclear factor κB (NF-κB) target genes were inhibited by fargesin treatment in the colon of the colitis mice. In vitro, fargesin blocked the nuclear translocation of p-p65, downregulated the protein levels of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), and dose-dependently inhibited the activity of NF-κB-luciferase in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Taken together, for the first time, the current study demonstrated the anti-inflammatory effects of fargesin on chemically induced IBD might be associated with NF-κB signaling suppression. The findings may contribute to the development of therapies for human IBD by using fargesin or its derivatives.

Flos Magnoliae and its Chemical Constituents Modulate Cl- Secretion via ANO1 Cl- Channel Inhibition in Human Airway Epithelial Cells

Flos Magnoliae and its Chemical Constituents Modulate Cl- Secretion via ANO1 Cl- Channel Inhibition in Human Airway Epithelial Cells

Flos Magnoliae Inhibits Chloride Secretion via ANO1 Inhibition in Calu-3 Cells.

Flos Magnoliae (FM, Chinese name: Xin-yi) is an oriental medicinal herb commonly used for symptomatic relief from allergic rhinitis, sinusitis, and headache, including in traditional Chinese and Korean medicine formulations. FM inhibits histamine release from mast cells and cytokine secretion from T cells. However, the mechanism of action of FM on the anoctamin-1 (ANO1) ion channel, which is responsible for nasal hypersecretion in allergic rhinitis, has not been elucidated. Therefore, in this study, we investigated the effect of a 30% ethanolic extract of FM (FMEtOH) and its chemical constituents on ANO1 activity. We used high-performance liquid chromatography analysis to identify five major chemical constituents of FMEtOH: vanillic acid, tiliroside, eudesmin, magnolin, and fargesin. Using a conventional whole-cell patch clamp method, we found that FMEtOH (30, 100, and 300[Formula: see text][Formula: see text]g/mL) and its chemical constituent tiliroside inhibited ANO1 activity in ANO1-overexpressing HEK293T cells. In addition, we found that the treatment of the airway epithelial cell line Calu-3 with interleukin 4 significantly increased Ca[Formula: see text] activated Cl[Formula: see text] current (ICaCC), but not cystic fibrosis transmembrane conductance regulator (CFTR)-mediated chloride current (ICFTR). FMEtOH and tiliroside specifically inhibited ICaCC. Thus, in this study, we identified a novel mechanism underlying the alleviation of allergic rhinitis by FMEtOH. Our results indicate that FMEtOH and its chemical constituent tiliroside are promising and potent agents for the prevention and treatment of allergic rhinitis.

Inhibitory Effects of Dimethyllirioresinol, Epimagnolin A, Eudesmin, Fargesin, and Magnolin on Cytochrome P450 Enzyme Activities in Human Liver Microsomes.

Magnolin, epimagnolin A, dimethyllirioresinol, eudesmin, and fargesin are pharmacologically active tetrahydrofurofuranoid lignans found in Flos Magnoliae. The inhibitory potentials of dimethyllirioresinol, epimagnolin A, eudesmin, fargesin, and magnolin on eight major human cytochrome P450 (CYP) enzyme activities in human liver microsomes were evaluated using liquid chromatography-tandem mass spectrometry to determine the inhibition mechanisms and inhibition potency. Fargesin inhibited CYP2C9-catalyzed diclofenac 4′-hydroxylation with a Ki value of 16.3 μM, and it exhibited mechanism-based inhibition of CYP2C19-catalyzed [S]-mephenytoin 4′-hydroxylation (Ki, 3.7 μM; kinact, 0.102 min−1), CYP2C8-catalyzed amodiaquine N-deethylation (Ki, 10.7 μM; kinact, 0.082 min−1), and CYP3A4-catalyzed midazolam 1′-hydroxylation (Ki, 23.0 μM; kinact, 0.050 min−1) in human liver microsomes. Fargesin negligibly inhibited CYP1A2-catalyzed phenacetin O-deethylation, CYP2A6-catalyzed coumarin 7-hydroxylation, CYP2B6-catalyzed bupropion hydroxylation, and CYP2D6-catalyzed bufuralol 1′-hydroxylation at 100 μM in human liver microsomes. Dimethyllirioresinol weakly inhibited CYP2C19 and CYP2C8 with IC50 values of 55.1 and 85.0 μM, respectively, without inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 activities at 100 μM. Epimagnolin A, eudesmin, and magnolin showed no the reversible and time-dependent inhibition of eight major CYP activities at 100 μM in human liver microsomes. These in vitro results suggest that it is necessary to investigate the potentials of in vivo fargesin-drug interaction with CYP2C8, CYP2C9, CYP2C19, and CYP3A4 substrates.

Pharmacokinetics and Tissue Distribution of Fargesin After Oral Administration in Rats by High Performance Liquid Chromatography

Background: Flos Magnoliae is a frequently-used Chinese herbal medicine, from which Fargesin is isolated with extensive pharmacological activities. Objective: To develop a reliable high performance liquid chromatography (HPLC) method of fargesin and explore pharmacokinetics and tissue distribution profile of fargesin in Sprague Dawley rats after oral administration of 50 mg/kg. Methods and Results: The chromatographic analysis was conducted on a Shimadzu 2010-ODS-3 C18 column (4.6 mmx150 mm, 5 µm), and the mobile phase consisted of methanol and water (58:42 v/v) with a flow rate of 1.0 mL/min. Linearity of fargesin in all biological samples was good (R>0.9990) within the corresponding concentration range and the method was verified to be sensitive, accuracy and precision. The pharmacokinetic results of fargesin after oral administration (50 mg/kg) in rats showed that two absorption peaks were observed in rat plasma at 60min and 290 min, with the highest plasma concentration (Cmax) being 464.38±32.75 ng/mL at 290 min. The tissue distribution results showed that the main tissue depots for fargesin were heart, liver, kidney and lung. Conclusion: This study established a sensitive and reliable HPLC method for the content determinations of fargesin. The pharmacokinetics and tissue distribution results may provide practical information for further study of pharmacological actions and clinical application. Keywords: Double-peak, Fargesin, HPLC, oral administration, pharmacokinetics, tissue distribution.

Biosynthesis of flat silver nanoflowers: from Flos Magnoliae Officinalis extract to simulation solution

Flat Ag nanoflowers were directly synthesized from the bioreduction of AgNO3 using Flos Magnoliae Officinalis extract without any additional stabilizer or protective agent at room temperature. Effects of concentrations of the Flos Magnoliae Officinalis extract on the Ag nanostructures were investigated. The main components containing flavone, polyphenol, protein, and reducing sugar in the plant extract were thoroughly determined before and after the reaction, and the dialysis experiments were also conducted. The results of components analysis and dialysis showed that gallic acid representing polyphenols played an important role in the biosynthesis of silver nanoplates. Trisodium citrate combined gallic acid solution, instead of Flos Magnoliae Officinalis extract, was employed and successfully simulated the biosynthesis process of the flat Ag nanoflowers.

English

The aim of this study was to study the anti-inflammatory effect of a compound containing volatile oils of&nbsp;Flos magnoliae,&nbsp;Centipeda&nbsp;minima&nbsp;and&nbsp;Mentha haplocalyx.&nbsp;The anti-inflammatory effect of the compound was investigated using xylene-, egg white- and cotton pellet-induced inflammation method.&nbsp;The&nbsp;medium- and high-doses of the compound&nbsp;(0.04&nbsp;and&nbsp;0.08 ml/kg) significantly inhibited the xylene-induced ear swelling in mice; low-, medium- and high-doses of the compound&nbsp;(0.02, 0.04,&nbsp;and&nbsp;0.08 ml/kg)&nbsp;all had significant inhibitory effects on degree of paw swelling 30 min after&nbsp;the induction of inflammation;&nbsp;high-dose of the compound (0.08 ml/kg)&nbsp;significantly lowered the weight of cotton pellet&nbsp;granuloma in rats.&nbsp;The compound has a preferable therapeutic effect on both the acute and chronic inflammations, which can be developed as an anti-inflammatory traditional Chinese medicine. &nbsp; Key words:&nbsp;Anti-inflammation,&nbsp;Flos magnoliae,&nbsp;Centipeda&nbsp;minima,&nbsp;Mentha haplocalyx, volatile oil.

PREPARATIVE SEPARATION AND PURIFICATION OF BIONDNOID I AND MAGNOLIN FROM THE CHINESE MEDICINAL PLANT FLOS MAGNOLIAE BY HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

A preparative high-speed counter-current chromatography (HSCCC) method for separation and purification of biondnoid I and magnolin from the Chinese medicinal plant Flos Magnoliae was successfully established by using light petroleum-ethyl acetate-methanol-water (3:7:4:6, v/v) as the two-phase solvent system. From 500 mg of crude extracts of Flos Magnoliae 48 mg of biondnoid I with a purity of 99.3% and 135 mg of magnolin with a purity of 98.7% were separated in a one-step separation. The structures of biondnoid I and magnolin were identified by 1H-NMR.

Fargesin, a component of Flos Magnoliae, stimulates glucose uptake in L6 myotubes

Flos Magnoliae (FM) is a commonly used Chinese medicinal herb for symptomatic relief of allergic rhinitis, sinusitis and headache. Although several FM species have been used as substitutes or adulterants for clinical use, possible differences in their pharmacological actions have not been reported. To confirm the effects of FM on skeletal muscle glucose metabolism, we tested the effects of several compounds isolated from FM on glucose uptake by L6 myotubes. We found that fargesin, a component of FM, dose-dependently stimulated glucose consumption in L6 myotubes, which was accompanied by enhanced glucose transporter (GLUT)-4 translocation to the cell surface. Fargesin-stimulated glucose uptake was blocked by wortmannin, a phosphatidylinositol-3 kinase (PI3 K) inhibitor. Fargesin stimulated Akt phosphorylation, a key component in the insulin signaling pathway, which was completely inhibited by wortmannin. Here, we demonstrated that fargesin, a bioactive component of Flos Magnoliae, increases basal glucose uptake and GLUT4 translocation in L6 myotubes by activating the PI3 K-Akt pathway.

Determination of the content of magnolin in Flos Magnoliae by microwave-assisted extraction and HPLC method

Objective: To establish a microwave-assisted extraction and HPLC method for determinination of the content of magnolin in Flos Magnoliae from four different origins.Methods: The magnolin in Flos Magnoliae herbs was extracted by 75% ethanol in microwave for 800 s at the temperature of 76 ℃.HPLC was performed on a Shim-Pack VP-ODS column(150 mm×4.6 mm,5 μm) with mixture of acetonitrile and water(40∶60) as mobile phase.The flow rate was 0.8 ml/min.The column temperature was room temperature.The detection wavelength was 238 nm.Results:The regression equation was A=2.922 7×104 c-3 617.4(r=0.999 9),where A was the peak areas and c was the concentration of magnolin.The concentrations of magnolin in the range of 5.6-240.8 μg/ml were in good linearity with chromatographic peak areas.The precision,reproducibility and stability were all good.The average recovery rates of low,middle and high concentrations(104.87,127.42,172.51 μg/ml) of magnolin were(100.39±1.58)%,(103.82±4.04)% and(103.74±0.13)%(n=3),respectively.This method was used to determine the contents of magnolin in Flos Magnoliae from four different origions: He'nan,Hebei,Hunan and Hubei provinces and the contents were(6.59±0.39)%,(3.44±0.48)%,(2.73±0.28)% and(4.18±0.25)%(n=3),respectively.Flos Magnoliae herbs from He'nan province was found to have the highest content of magnolin.Conclusion: The method is simple,accurate,stable and can be used as a quantitative method for quality control of Flos Magnoliae herbs and its preparations.

Antimicrobial Activity of Traditional Chinese Medicines on Common Oral Bacteria

Objective: To evaluate twenty Traditional Chinese Medicines (TCM) against four oral bacteria. Methods: Twenty TCM were tested for sensitivity against Streptococcus mitis, Streptococcus sanguis, Streptococcus mutans and Porphyromonas gingivalis. Aliquots of suspension of each bacterial species were inoculated on a horse blood agar (HBA) plate, 6 mm diameter paper disks was soaked in different drug suspensions were placed concentrically on a HBA plate. Disks soaked in 0.2% w/v chlorhexidine were used as positive controls. These HBA plates were incubated for 48 hours anaerobically and the diameters of growth inhibition of three different areas were measured using a calibrated computer software and the mean diameter obtained for each bacteria. Broth microdilution assay was used to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The experiment was repeated on three separate occasions. Results: The TCMs that consistently against Porphyromonas gingivalis, included Folium artemisiae argyi, Fructus crataegi, Rhizoma dryopteris crassirhizomae, Flos magnoliae, Rhizoma polygoni cuspidati, Radix scrophulariae ningpoensis, Galla chinensis, Radix scutellariae baicalensis and Rhizoma coptidis; against Streptococcus mutans included Fructus crataegi, Galla chinensis and Rhizoma copitidis; against Streptococcus mitis and Streptococcus sanguis included Galla chinensis and Rhizoma copitidis. Conclusion: Rhizoma copitidis and Galla chinensis had inhibitory effects on Streptococcus mitis, Streptococcus sanguis, Streptococcus mutans and Porphyromonas gingivalis in vitro.