wogonoside and wogonin

Nor-wogonin, a polyhydroxy flavone, has been shown to possess antitumor activity. However, the mechanisms responsible for its antitumor activity are poorly studied. Herein, we investigated the mechanisms of nor-wogonin actions in triple-negative breast cancer (TNBC) cells.. Effects of nor-wogonin on cell proliferation and viability of four TNBC cell lines (MDA-MB-231, BT-549, HCC70, and HCC1806) and two non-tumorigenic breast cell lines (MCF-10A and AG11132) were assessed by BrdU incorporation assays and trypan blue dye exclusion tests. Cell cycle and apoptosis analyses were carried out by flow cytometry. Protein expression was analyzed by immunoblotting.. Nor-wogonin significantly inhibited the growth and decreased the viability of TNBC cells; however, it exhibited no or minimal effects in non-tumorigenic breast cells. Nor-wogonin (40 μM) was a more potent anti-proliferative and cytotoxic agent than wogonin (100 μM) and wogonoside (100 μM), which are structurally related to nor-wogonin. The antitumor effects of nor-wogonin can be attributed to cell cycle arrest via reduction of the expression of cyclin D1, cyclin B1, and CDK1. Furthermore, nor-wogonin induced mitochondrial apoptosis, (as evidenced by the increase in % of cells that are apoptotic), decreases in the mitochondrial membrane potential (ΔΨm), increases in Bax/Bcl-2 ratio, and caspase-3 cleavage. Moreover, nor-wogonin attenuated the expression of the nuclear factor kappa-B and activation of signal transducer and activator of transcription 3 pathways, which can be correlated with suppression of transforming growth factor-β-activated kinase 1 in TNBC cells.. These results showed that nor-wogonin might be a potential multi-target agent for TNBC treatment.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Down-Regulation; Female; Flavanones; Flavones; Glucosides; Humans; MAP Kinase Kinase Kinases; NF-kappa B; STAT3 Transcription Factor; Triple Negative Breast Neoplasms

Scutellaria baicalensis is commonly used in Asia as an herbal medicine to treat a variety of ailments, including cancer. Wogonoside, one major constituent of S. baicalensis, can be primarily converted to wogonin through deglycosylation via enteric microbiome metabolism.. The antiproliferative effects of the glycoside (wogonoside) and its deglycosylated compound (wogonin) on a panel of human cancer cell lines from the most common solid tumors were evaluated using the MTS colorimetric assay. Cell cycle and apoptosis were determined using flow cytometry. Enzymatic activities of caspases were measured, and the interactions of wogonin and caspases were explored by a docking analysis.. Wogonoside did not have obvious antiproliferative effects on the cancer cells. In contrast, wogonin showed significant antiproliferative activities on all the tested cancer cells. Wogonin arrested the cells in the G1 phase and significantly induced cell apoptosis. The compound also activated the expression of caspases 3 and 9. The docking results suggest that the compound forms hydrogen bonds with Phe250 and Ser251, and π-π interactions with Phe256 in caspase 3, and with Asp228 in caspase 9.. After wogonoside deglycosylation, wogonin significantly enhanced its anticancer potential as a potent anticancer compound derived from S. baicalensis.

Topics: Apoptosis; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Proliferation; Flavanones; Flow Cytometry; Gene Expression Regulation, Neoplastic; Glucosides; Glycosylation; Humans; Hydrogen Bonding; MCF-7 Cells; Microbiota; Molecular Docking Simulation; Neoplasms; Phytotherapy; Plant Extracts; Scutellaria baicalensis

Scutellariae Radix (SR) and its bioactive flavones elicit a variety of effects in the brain. However, the brain uptake of individual SR flavones and its relationship to the elicited effects after SR administration remain unknown. Moreover, previous studies seldom measured pharmacokinetic and pharmacodynamic outcomes simultaneously. In the current study, the brain uptake of six major SR flavones and the anxiolytic behavior following oral administration of a SR extract at two clinically relevant doses (600 and 1200 mg/kg twice daily) were simultaneously investigated in mice (n = 18 per group). Brain and plasma concentrations of the flavones were measured by LC-MS/MS, while the anxiolytic effect was evaluated using the elevated plus maze. To further investigate the mechanism behind the differential brain uptake of the six SR flavones, these flavones were separately administered to mice at an equivalent molar oral dose (n = 6). The brain tissue bindings of the SR flavones were also measured with the in vitro brain slice method. Our results indicated that all six SR flavones including three aglycons (baicalein, wogonin, and oroxylin A) and three glucuronides (baicalin, wogonoside, and oroxyloside) could pass through the blood-brain barrier, with brain concentrations ranging from 7.9 to 224.0 pmol/g. It provided novel evidence that oroxylin A had the highest brain uptake among the six SR flavones regardless of its limited content in SR extract, in which 3.6-3.9% of the administered oroxylin A dose was present in the brain 6 h postdosing and with a brain-to-plasma ratio of 0.42-0.46. Although SR extract contains flavones that are positive modulators of the benzodiazepine binding site of GABA

Topics: Animals; Anti-Anxiety Agents; Brain; Flavanones; Flavones; Flavonoids; Glucosides; Male; Maze Learning; Mice; Mice, Inbred ICR; Scutellaria baicalensis; Tandem Mass Spectrometry

The growing interest in skin lightening has recently renewed attention on the esthetic applications of Chinese herbal medicine. Although Scutellaria baicalensis Georgi is used for antipyretic and antiinflammatory purposes, its whitening effect remains unclear. This study reports three major findings: (1) S. baicalensis has a potent inhibitory effect on melanogenesis; (2) wogonin and its glycoside are the active components of S. baicalensis; and (3) O-methylated flavones from S. baicalensis, such as wogonin, inhibit intracellular melanosome transport. Using a melanin quantification assay, we showed that S. baicalensis potently inhibits melanogenesis in B16F10 cells. Componential analyses revealed that the main components of S. baicalensis are baicalin, wogonoside, baicalein, wogonin, and oroxylin A. Among these five flavones, wogonin and wogonoside consistently inhibited melanogenesis in both B16F10 melanoma cells and primary melanocytes. Wogonin exhibited the strongest inhibition of melanin production and markedly lightened the color of skin equivalents. We identified microphthalmia-associated transcription factor and tyrosinase-related proteins as potential targets of wogonin- and wogonoside-induced melanogenesis suppression. In culture, we found that the melanosomes in wogonin-treated B16F10 cells were localized to the perinuclear region. Immunoblotting analyses revealed that wogonin significantly reduced in melanophilin protein, which is required for actin-based melanosome transport. Other actin-based melanosome transport-related molecules, i.e., Rab27A and myosin Va, were not affected by wogonin. Cotreatment with MG132 blocked the wogonin-induced decrease in melanophilin, suggesting that wogonin promotes the proteolytic degradation of melanophilin via the calpain/proteasomal pathway. We determined that the structural specificities of the mono-O-methyl group in the flavone A-ring and the aglycone form were responsible for reducing melanosome transport. Furthermore, wogonin and two wogonin analogs, mono-O-methyl flavones, strongly suppressed melanosome transport. Our findings suggest the applicability of S. baicalensis in the esthetic field. Thus, we propose a novel pharmacologic approach for the treatment of hyperpigmentation.

Topics: Animals; Biological Transport; Cell Line, Tumor; Cells, Cultured; Flavanones; Glucosides; Humans; Melanins; Melanocytes; Melanosomes; Mice; Plant Extracts; Proteasome Endopeptidase Complex; Scutellaria baicalensis; Transcription Factors

This paper is aimed to study plant biomass and active compounds of Scutellaria baicalensis germchit in different five stages (from germination to transplant). The length of shoot and root, the diameter and the weight of root were determined. HPLC method was used to determine the content of active compounds (baicalin, scutellarin, wogonoside, baicalein, wogonin). According to the results, various biological indicators increased with the germination of seedling. However, the drying rate of the root declined to 27.96% from 32.90%. The contents of scutellarin and baicalein increased firstly, and then decreased. The maximums of them were 3.22,3.89 mg•g⁻¹ while the data of shoot/root was 0.35. The maximums of the contents of baicalin and wogonoside were 107.39,16.11 mg•g⁻¹ while the data of shoot/root was 0.23 and 0.06. The contents of wogonin gradually increased to the maximum of 0.88 mg•g⁻¹ while the data of shoot/root was 0.50. In conclusion, the contents of baicalin, scutellarin, wogonoside, baicalein and wogonin reached or approached the maximum at germination stage while the data of shoot/root was 0.35. The rate of shoot and root can be used as a judging index of active compounds for S. baicalensis germchit.

Topics: Biomass; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Germination; Glucosides; Plant Extracts; Plant Roots; Scutellaria baicalensis; Seedlings

A simple and accurate high-performance liquid chromatography-photodiode array (HPLC-PDA) detection method has been developed and validated for simultaneous determination of nine components-liquiritin, coptisine, baicalin, palmatine, berberine, wogonoside, baicalein, glycyrrhizin and wogonin-in the traditional Korean formula, Banhasasim-tang decoction. A Gemini C18 analytical column was used to separate the nine constituents and kept at 40°C by gradient elution with 0.1% (v/v) trifluoroacetic acid in distilled water (A) and acetonitrile (B) as mobile phases. The flow rate was 1.0 mL/min and the injection volume was 10 µL. The PDA detection wavelengths were set at 254, 275 and 350 nm. Calibration curves of all compounds showed good linearity with coefficients of determination ≥0.9998 within the test ranges. The limits of detection and quantification of all compounds were in the range 0.01-0.09 and 0.03-0.30 µg/mL, respectively. All recoveries of the nine marker compounds ranged from 98.65 to 103.22% with relative standard deviation (RSD) values <1.25%. The RSDs of intraday and interday precision were <1.13 and 1.83%, respectively. The concentrations of the nine marker constituents were 0.19-41.09 mg/g.

Topics: Acetonitriles; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Glucosides; Glycyrrhizic Acid; Humans; Limit of Detection; Medicine, Korean Traditional; Observer Variation; Pinellia; Plants, Medicinal; Reproducibility of Results; Solvents; Trifluoroacetic Acid; Water

Chlorophyll-type contaminants are commonly encountered in the isolation and determination of flavones of plant aerial plant parts. Heme is also a difficult background substance in whole blood analysis. Both chlorophyll and heme are porphyrin type compounds. In this study, a rapid method for isolating flavones with 5-hydroxyl or ortho-hydroxyl groups from biological samples was developed based on the different solubilities of porphyrin-metal and flavone-metal complexes. It is important that other background substances, e.g., proteins and lipids, are also removed from flavones without an additional processing. The recoveries of scutellarin, baicalin, baicalein, wogonoside and wogonin, which are the primary constituents of Scutellaria baicalensis (skullcaps) were 99.65% ± 1.02%, 98.98% ± 0.73%, 99.65% ± 0.03%, 97.59% ± 0.09% and 95.19% ± 0.47%, respectively. As a sample pretreatment procedure, this method was coupled to high-performance liquid chromatography (HPLC) with good separation, sensitivity and linearity and was applied to determine the flavone content in different aerial parts of S. baicalensis and in dried blood spot samples.

Topics: Apigenin; Chromatography, High Pressure Liquid; Dried Blood Spot Testing; Flavanones; Flavones; Flavonoids; Glucosides; Glucuronates; Humans; Plant Components, Aerial; Plant Extracts; Scutellaria baicalensis; Zinc Acetate

We evaluated the antioxidant properties of four main flavones from Scutellaria baicalensis: baicalein, wogonin and their glucuronides - baicalin and wogonoside. We used three in vitro assays: free radical scavenging with 2,2'-diphenylpicrylhydrazyl radical, transition metal ions reducing power by phosphomolybdenum assay and inhibition of the hydroxyl radical-induced peroxidation of linoleic acid assay. All flavones have antioxidant capacity, which differs depending on the structure and mechanisms of activity. In all tests, only baicalein - the aglycone with three adjacent hydroxyl groups - exhibited consistent antioxidant effect. Wogonin protected linoleic acid against oxidation. Baicalin displayed less potent antioxidant properties whereas wogonoside did not have significant antioxidant activity.

Topics: Antioxidants; Flavanones; Flavones; Flavonoids; Free Radical Scavengers; Glucosides; Molecular Structure; Plant Extracts; Plant Roots; Scutellaria baicalensis

This study aims to identify and quantify the six major bioactive flavones of the traditional Chinese medicine Scutellariae Radix (RS), including baicalein, baicalin, wogonin, wogonoside, oroxylin A and oroxyloside in rat after oral administration of a standardized RS extract. A novel, sensitive and selective method for simultaneous determination of these six analytes in rat brain and plasma using solid phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC/MS/MS) was developed and fully validated. The lower limits of quantification (LLOQs) for the six RS flavones in brain tissue were 0.02nmol/g. The LLOQs in plasma were 0.005nmol/ml for B, W and OA, 0.025nmol/ml for WG and OAG, and 0.1875nmol/ml for BG. The current study provides novel evidence of the presence of all the tested RS flavones and an isoform of BG (BG', probably baicalein-6-O-glucuronide) in the rat brain after oral administration of RS extract, suggesting their ability to permeate through the blood-brain barrier. The method was also successfully applied to the pharmacokinetic study of all these analytes in plasma after oral administration of RS extract (300mg/kg) to Sprague-Dawley rats. The developed assay method provides a useful tool for both preclinical and clinical investigations on the disposition of RS flavones in brain and plasma.

Topics: Animals; Brain; Chromatography, Liquid; Flavanones; Flavones; Flavonoids; Glucosides; Glucuronides; Male; Rats; Solid Phase Extraction; Tandem Mass Spectrometry

Wogonin (WGN), a flavonoid extracted from Scutellaria baicalensis Georgi, has several biological effects including antioxidant, anti-inflammatory, antiviral, neuroprotective, anxiolytic, and anticancer activities, and the flavonoid wogonoside (WGNS) can be derived from S. baicalensis, as it is a metabolite of wogonin. Here, the anticoagulant activities of WGN(S) were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin (factor IIa, FIIa) and activated factor X (FXa), and the effects of WGN(S) on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-α activated human umbilical vein endothelial cells (HUVECs). Treatment with WGN(S) resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, WGN(S) inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. WGN(S) also elicited anticoagulant effects in mice. In addition, treatment with WGN(S) resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, WGN(S) possesses antithrombotic activities and offers a basis for development of a novel anticoagulant.

Topics: Adult; Animals; Anticoagulants; Factor Xa; Female; Flavanones; Glucosides; Human Umbilical Vein Endothelial Cells; Humans; Male; Mice; Mice, Inbred ICR; Partial Thromboplastin Time; Plasminogen Activator Inhibitor 1; Platelet Aggregation; Platelet Aggregation Inhibitors; Prothrombin Time; Scutellaria baicalensis; Thrombin

The pharmacodynamic (PD) and pharmacokinetic (PK) properties of Huangqin Tang (HQT) were investigated in yeast-induced febrile rats. Blood sample and rectal temperature data of the rats were collected at different times after single oral administration of HQT at 20 g x kg(-1). The plasma concentrations of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid were quantified by a sensitive liquid chromatography-tandem mass spectrometric (LC-MS) method. The blood concentrations of PGE2, 1L-1β and TNF-α were detected by radioimmunoassay (RIA). All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The potential relationship between the mean concentration of eight constituents and the antifebrile efficacy was investigated by calculating Pearson correlation coefficients. It was found that HQT had significant antifebrile efficacy in yeast-induced febrile rats, but had no effect to normal rats. The antifebrile effect of HQT can be attributed to the inhibition of PGE2, 1L-1β and TNF-α. The constituents (baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid) in febrile rats had delayed absorption and elimination, a longer residence time in the body, and higher C(max) and AUC than those in normal rats. Febrile condition could affect the pharmacokinetic behaviour of HQT in vivo; the flavonoids with the same backbone showed the similar fate in the body; baicalein and wogonin had a strong positive correlation (R > 0.66, P ≤ 0.02) with the antifebrile efficacy determined. Together, these constituents demonstrated different pharmacokinetic properties in the febrile body.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, Liquid; Dinoprostone; Drugs, Chinese Herbal; Fever; Flavanones; Flavonoids; Glucosides; Interleukin-1beta; Mass Spectrometry; Monoterpenes; Rats; Tumor Necrosis Factor-alpha

Scutellaria baicalensis Georgi (Labiatae) is one of the most commonly used medicinal herbs, especially in traditional Chinese medicine. However, compared to many pharmacological studies of this botanical, much less attention has been paid to the quality control of the herb's pretreatment prior to extract preparation, an issue that may affect therapeutic outcomes.. The current study was designed to evaluate whether different pretreatment conditions change the contents of the four major flavonoids in the herb, i.e., two glycosides (baicalin and wogonoside) and two aglycones (baicalein and wogonin).. A high-performance liquid chromatography assay was used to quantify the contents of these four flavonoids. The composition changes of four flavonoids by different pretreatment conditions, including solvent, treatment time, temperature, pH value and herb/solvent ratio were evaluated.. After selection of the first order time-curve kinetics, our data showed that at 50 °C, 1:5 herb/water (in w/v) ratio and pH 6.67 yielded an optimal conversion rate from flavonoid glycosides to their aglycones. In this optimized condition, the contents of baicalin and wogonoside were decreased to 1/70 and 1/13, while baicalein and wogonin were increased 3.5- and 3.1-fold, respectively, compared to untreated herb.. The markedly variable conversion rates by different pretreatment conditions complicated the quality control of this herb, mainly due to the high amount of endogenous enzymes of S. baicalensis. Optimal pretreatment conditions observed in this study could be used obtain the highest level of desired constituents to achieve better pharmacological effects.

Topics: Chemical Fractionation; Chromatography, High Pressure Liquid; Drug Stability; Flavanones; Flavonoids; Glucosides; Hydrogen-Ion Concentration; Kinetics; Phytotherapy; Plant Extracts; Plants, Medicinal; Quality Control; Scutellaria baicalensis; Solvents; Temperature

Scutellaria baicalensis has been used as a traditional herbal medicine for bronchitis, hepatitis, and allergic diseases. The root of Scutellaria baicalensis contains active flavonoid components, including baicalin, baicalein, wogonoside, and wogonin, which have pharmaceutical properties. In the present study, the antiallergic properties of a standardized aqueous extract of S. baicalensis were evaluated, and the skin toxicity of its dermal application was also determined. The in vivo and in vitro assays were performed by using the β-hexosaminidase assay in rat basophilic leukemia cells (RBL-2H3) and cutaneous skin reaction in BALB/c mice, respectively. In addition, the acute dermal irritation/corrosion test was carried out in New Zealand white rabbits, and the skin sensitization test was conducted by Buhler's method in Hartley guinea pigs to estimate the safety of the standardized aqueous extract of S. baicalensis for topical application. β-Hexosaminidase release in RBL-2H3 was markedly decreased following treatment with the standardized aqueous extract of S. baicalensis. It also ameliorated antigen-induced ear swelling compared with the control group in BALB/c mice. In the toxicological studies, it did not induce any dermal irritation/corrosion in rabbits or skin sensitization in guinea pigs. Although still limited, these results concerning the toxicological effects of S. baicalensis could be an initial step toward the topical application of S. baicalensis extracts on hypersensitive skin.

Topics: Administration, Topical; Animals; Anti-Allergic Agents; beta-N-Acetylhexosaminidases; Cell Line, Tumor; Flavanones; Flavonoids; Glucosides; Guinea Pigs; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Plant Extracts; Plant Roots; Plants, Medicinal; Rabbits; Rats; Scutellaria baicalensis; Skin

The current study aims to investigate the pharmacokinetic properties of Huangqin Tang on different oral doses. An LC-MS method for simultaneous determination of flavonoids and terpenoids in rat plasma was developed and validated. Plasma samples were treated with hydrochloric acid (containing 1% ascorbic acid), precipitated with acetonitrile, separated on a Zorbax SB-C18 column, detected by single quadruple mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. All pharmacokinetic parameters were processed by non-compartmental analysis using WinNonlin software. The results of specificity, linearity, intra-day and inter-day precisions, accuracy, and stability for LC-MS assay were suitable for the quantification of paeoniflorin, baicalin, wogonoside, baicalein, wogonin, oroxylin A, glycyrrhizic acid and glycyrrhetinic acid in rat plasma. The concentration-time profiles of baicalin, wogonoside, baicalein, wogonin, oroxylin A and glycyrrhizic acid showed double-peak phenomenon after Huangqin Tang was orally administered at 40 g x kg(-1) dose; all eight constituents in rat plasma showed good dose-exposure relationship within the dosage of 10-40 g x kg(-1); although plasma concentrations were different, the flavonoids with the same backbone showed the similar fate in the body with the corresponding dosage. In conclusion, the LC-MS assay was successfully applied for the pharmacokinetic study of multi-constituents of Huangqin Tang with different doses. Additionally, these constituents demonstrated good pharmacokinetic properties in the body.

Topics: Administration, Oral; Animals; Chromatography, Liquid; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Male; Monoterpenes; Pentacyclic Triterpenes; Rats; Rats, Wistar; Spectrometry, Mass, Electrospray Ionization

Scutellariae Radix, the dried root of Scutellaria baicalensis Georgi, has been widely used in Asian countries for the treatment of dermatitis, diarrhoea, inflammatory disease and hepatic disease. A simple, sensitive and precise reversed-phase liquid chromatographic method with isocratic elution was developed to simultaneously determine four bioactive compounds in Scutellariae Radix: baicalein, baicalin, wogonin and wogonoside. Chromatographic analysis was performed on a YMC Pack Pro C(8) column (150 × 4.6 mm(2), 3 µm), with a mobile phase of 0.1% formic acid : acetonitrile (70 : 30, v/v) at a flow rate of 1.0 mL min(-1), and UV detection at 280 nm. Linear behaviour was observed over the investigated concentration range (0.25-10 µg mL(-1)) for all analytes, with a correlation coefficient of >0.997. The intra- and inter-day precisions were <8.07%, and accuracies were 92.3-102.9%. This method was successfully applied for the analysis of marker compounds for the quality control of Scutellariae Radix extract.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Limit of Detection; Quality Control; Reproducibility of Results; Scutellaria baicalensis

Reversed phase high performance liquid chromatography with diode array detector was employed for simultaneous determination of five main index components and specific chromatograms analysis in Xiaochaihu granules with a linear gradient elution of acetonitrile-water (containing 0.1% phosphoric acid) as mobile phase. The results showed that five main index components (baicalin, baicalein, wogonoside, wogonin, enoxolone) were separated well under the analytical condition. The linear ranges of five components were 0.518 - 16.576, 0.069 - 2.197, 0.167 - 5.333, 0.009 - 0.297 and 0.006 - 0.270 mg x g(-1), respectively. The correlation coefficients were 0.999 9, and the average recoveries ranged from 95% to 105%. Twelve common peaks were selected as the specific chromatograms of Xiaochaihu granules with baicalin as the reference peak. There were good similarities between the reference and the ten batches of samples. The similarity coefficients were no less than 0.9. The analytical method established is highly sensitive with strong specificity and it can be used efficiently in the quality control of Xiaochaihu granules.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhetinic Acid; Quality Control; Sensitivity and Specificity

This work is to study the baicalin and its three analogs, baicalin, wogonoside, and wogonin, on the protective effect of neuron from oxygen-glucose deprivation (OGD) and toll-like receptor 2 (TLR2) expression in OGD damage. The results showed that baicalin and its three analogs did protect neurons from OGD damage and downregulated protein level of TLR2. D-Glucopyranosiduronic acid on site 7 in the structure played a core of cytotoxicity of these flavonoid analogs. The methoxyl group on carbon 8 of the structure had the relation with TLR2 protein expression, as well as the anti-inflammation. In addition, we detected caspase3 and antioxidation capability, to investigate the effect of four analogs on cell apoptosis and total antioxidation competence in OGD model.

Topics: Animals; Antioxidants; Caspase 3; Cell Survival; Flavanones; Flavonoids; Glucose; Glucosides; Immunity, Innate; Neurons; Oxygen; PC12 Cells; Rats; Toll-Like Receptor 2; Tumor Necrosis Factor-alpha

A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of baicalin, wogonoside, baicalein, wogonin, oroxylin A and chrysin in rat plasma, using naringin as an internal standard. After acidifying with HCl, plasma samples were pretreated by liquid-liquid extraction with acetone. Chromatographic separation was accomplished on a Hypersil Gold-C(18) analytical column (2.1×150 mm, 5 μm) utilizing a gradient elution profile and a mobile phase consisting of (A) 0.1% formic acid in water and (B) acetonitrile. Detection was performed by multiple reaction monitoring (MRM) mode using electrospray ionization in the positive ion mode. All analytes showed good linearity over the investigated concentration range (r>0.9900). The lower limit of quantification was 0.5 ng/ml for baicalin, wogonoside, wogonin and oroxylin A, and 1.0 ng/ml for baicalein and chrysin. Intra-day and inter-day precisions (RSD%) were less than 15% and accuracy (RE%) ranged from -6.7% to 5.8%. The validated method was successfully applied to investigate the pharmacokinetics of the major flavonoids of Radix scutellariae extract after oral administration to rats.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Half-Life; Limit of Detection; Male; Metabolic Clearance Rate; Plant Extracts; Rats; Rats, Wistar; Reference Standards; Reproducibility of Results; Scutellaria baicalensis; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

To study the transportation of Xiaochaihu Tang in Caco-2 cell model.. The safety concentration of Xiaochaihu Tang in Caco-2 cells was determined by MTT assay. Then the Caco-2 cell model was used to investigate the bi-directional transportation of Xiaochaihu Tang. The multicomponents of Xiaochaihu Tang and the influence of time were measured by high performance liquid chromatography (HPLC).. The P(app) values of wogonoside and wogonin were (1.23 +/- 0.09) x 10(-6), (1.07 +/- 0.89) x 10(-5) cm x s(-1) from the AP side to BL side, and (2.12 +/- 0.19) x 10(-6) and (7.12 +/- 1.02) x 10(-6) cm x s(-1) from the BL side to AP side, respectively. The P(appAP --> BL)/P(app BL --> AP) ratio of wogonoside and wogonin were 0.58 and 1.49, respectively. Baicalin, baicalein and glycyrrhizic acid could not permeate the Caco-2 cell model.. The transportation of wogonoside and wogonin in Caco-2 cell model may be a passive transportation.

Topics: Biological Transport, Active; Caco-2 Cells; Cell Line, Tumor; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhizic Acid; Humans

To establish a HPLC-PDA method for the determination of baicalin, wogonoside, baicalein, wogonin and glycyrrhizic acid in Xiaochaihu Tang.. A Symmetry Shield RP18 (4.6 mm x 250 mm, 5.0 microm) was used with a mobile phase of acetonitrile-0.01% H3PO4 in gradient elution. The detection wavelength was 251 nm,the flow rate was 0.45 mL x min(-1) and the column temperature was maintained at 30 degrees C.. The accuracy, precision, sensitivity, specificity and linearity of this method met the requirements. The contents of the five effective fractions were determined simultaneously.. The method is rapid,simple and accurate and it can be suitable for the determination of baicalin, wogonoside, baicalein, wogonin and glycyrrhizic acid in Xiaochaihu Tang simultaneously.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhizic Acid; Indicators and Reagents; Medicine, Chinese Traditional; Rehmannia; Spectrophotometry, Ultraviolet; Technology, Pharmaceutical

A rapid, sensitive, and specific method by high-performance liquid chromatography (HPLC) coupled to diode-array detection (DAD) and tandem mass spectrometry (MS) techniques was developed for the identification of absorbed constituents and their metabolites in rats after the oral administration of a Chai-Huang decoction (CHD), which consists of Bupleurum chinense and Scutellaria baicalensis in the proportion 1 : 1 (w/w). By comparing their retention times and MS data with those of authentic compounds and published data, a total of 14 compounds were identified in the CHD samples. In addition, eleven and seven compounds were characterized in the urine and serum samples of the rats, respectively. The results indicated that the main absorbed constituents were chrysin-6-C-arabinosyl-8-C-glucoside, chrysin-6-C-glucosyl-8-C-arabinoside, baicalin, wogonin-5-O-glucoside, oroxylin A-7-O-glucuronide, wogonoside, saikosaponin A, saikosaponin C, saikosaponin D, baicalein, and wogonin. These compounds might be responsible for the curative effects of the CHD. The findings demonstrated that the proposed method could be used to rapidly and simultaneously analyze and screen the multiple absorbed bioactive constituents in a formula of traditional Chinese medicines (TCM). This is very important not only for the pharmaceutical discovery process and the quality control of crude drugs but also to explain the mechanisms of action of TCM.

Topics: Administration, Oral; Animals; Bupleurum; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glucuronides; Medicine, Chinese Traditional; Oleanolic Acid; Rats; Saponins; Scutellaria baicalensis; Spectrometry, Mass, Electrospray Ionization

The beta-glucuronidase (GUS) gene from Lactobacillus brevis RO1 was cloned and expressed in Escherichia coli GMS407. The GUS gene was composed of 1812 bp, encoding a 603-amino-acid protein belonging to the glycosyl hydrolase family 2 with three conserved domains. The amino acid similarity was higher than 70% with the beta-glucuronidases of various microorganisms, yet less than 58% with the beta-glucuronidase of L. gasseri ADH. Overexpression and purification of the GUS was performed in beta-glucuronidase-deficient E. coli GMS407. The purified GUS protein was 71 kDa and showed 1284 U/mg of specific activity at optimum condition of pH 5.0 and 37 degrees C. At 37 degrees C, the GUS remained stable for 80 min at pH values ranging from 5.0 to 8.0. The purified enzyme exhibited a half-life of 1 h at 60 degrees C and more than 2 h at 50 degrees C. When the purified GUS was applied to transform baicalin and wogonoside into their corresponding aglycones, 150 microM of baicalin and 125 microM of wogonoside were completely transformed into baicalein and wogonin, respectively, within 3 h.

Topics: Amino Acid Sequence; Cloning, Molecular; Enzyme Stability; Escherichia coli; Flavanones; Flavonoids; Glucosides; Glucuronidase; Hydrogen-Ion Concentration; Levilactobacillus brevis; Molecular Sequence Data; Molecular Weight; Sequence Homology, Amino Acid; Substrate Specificity; Temperature; Time Factors

To develop a quantitative method for simultaneously determining four flavones in Radix Scutellariae, by using one chemical standard substance.. The relative correction factors (RCF) of four flavones were determined by HPLC-DAD. Within the linear ranges, the values of RCF at 274 nm of wogonoside, baicalein, and wogonin to baicalin, were 0.7467, 0.5638, 0.4351, respectively. And the contents of baicalin in 27 samples of Scutellaria baicalensis were authentically determined by the external standard method, and the contents of the three other flavones were calculated according to their RCF. The contents of these four flavones in the all samples were determined with the external standard method.. The RCF had a good reproducibility (RSD 0.66%-2.83%). No significant differences between the quantitative results by the above two methods were observed.. It is a fast, convenient, and accurate method to determine multi-components especially when some authentic standard substances were unavailable. It can be used as a method to control the quality of Radix Scutellariae.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavones; Flavonoids; Glucosides; Plant Extracts; Scutellaria baicalensis

Scutellaria baicalensis was collected from four wild and four cultivated populations from different locations in China. Forty-two samples were analyzed using random amplification of polymorphic DNA (RAPD) techniques for genetic profiling, and high performance liquid chromatography (HPLC) techniques to determine the flavonoid content. The selected 23 RAPD primers yielded a total of 838 clear and reproducible bands of which 237 were found to be polymorphic. The wild population exhibited higher polymorphism than that of the cultivated population. The dendrogram generated by the UPGMA method via Nei's genetic distance revealed three distinct genotypes from the cultivated populations and several branches from the wild populations. The contents of baicalin and wogonoside in dried roots of the samples ranged from (w/w) 8.63 to 17.84%, and from 1.99 to 4.21%, respectively, whereas their aglycones, baicalein and wogonin, were within the range of only 0.04-0.23%. The total content of the four flavonoids varied from 9.45 to 26.24%. Comparatively, the cultivated populations contained much higher levels of baicalin and total flavonoids than those in the wild populations. The results from genetic characterization and phytochemical analysis demonstrated that the quality variation of this drug was mainly determined by extrinsic environmental or agricultural factors, rather than by genetic differences. Our findings can be used for the commercial production and germplasm management of this medicinal plant.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Genetic Variation; Glucosides; Random Amplified Polymorphic DNA Technique; Scutellaria baicalensis

To study the pharmacokinetics of flavonoids from Xiexin decoction in rats. SD rats were given a single ig dose of Xiexin decoction 12 g x kg(-1), plasma and urine were collected before and after dosing. Flavonoids components in plasma and urine were measured by HPLC. Pharmacokinetic parameters were determined from the plasma concentration-time data and urinary excretion-time data with the DAS software package. Baicalin was incubated with the rat renal homogenate to investigate its metabolism in vitro. After oral administration of Xiexin decoction baicalin and wogonoside were quickly absorbed and exhibited double peak phenomena in their plasma concentrations. The first peaks in plasma concentrations of baicalin and wogonoside reached Cmax1 of (10 +/- 8) and (1.5 +/- 0.5) mg x L(-1) at Tmax1 of (0.27 +/- 0.09) and (0.17 +/- 0.00) h, while the second peaks reached Cmax2 of (3. 9 0. 5) and (0. 74 +/- 0.11) mg x L(-1) at Tmax2 of (7.6 +/- 2.6) and (16.0 +/- 0.0) h, respectively. The T(1/2) of baicalin and wogonoside were (7 +/- 3) and (6.4 +/- 2.1) h, AUC(0-infinity) were (57 +/- 12) and (15 +/- 3) mg x h x L(-1), respectively. After oral administration of Xiexin decoction, not only baicalin and wogonoside but also baicalein and wogonin can be detected in the urine. The amounts of baicalin, wogonoside, baicalein and wogonin excreted from urine during 0-72 h were (1.4 +/- 0.3), (3.4 +/- 1.3), (2.2 +/- 0.97), (10 +/- 4)% of dose given in rats, respectively. The excretion T(1/2) of the four flavonoids were (6.9 +/- 2.1), (9 +/- 4) , (8.2 +/- 2.0) and (7.2 +/- 1.8) h, respectively. Baicalin was metabolized into baicalein in the rat renal homogenate in vitro, and the kinetic parameters were measured as Vmax = 702 nmol x min(-1) x g(-1) (protein) and Km=135 micromol x L(-1). After oral administration of Xiexin decoction, flavonoids can be absorbed quickly. Only a small quantity of baicalin, wogonoside, baicalein and wogonin were excreted from urine. Baicalin may be metabolized into baicalein in the rat kidney.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Kidney; Male; Rats; Rats, Sprague-Dawley

To establish the HPLC fingerprint of the flavonoids of Kushen Tang, and to study its correlation to Scutellaria baicalensis and Sophora flavescens.. The fingerprints of the flavonoids of Kushen Tang were established by HPLC under two detective wavelengths 280 nm and 365 nm. The correlation existed in the common peaks of HPLC fingerprint of Kushen Tang and the Chinese meteria medicines which composed the prescription was explored by analyzing the HPLC fingerprints of different compatibilities. The chemical constituents which the common peaks stood for were analyzed by adding controls to the samples respectively.. Under the detective wavelength 280 nm, 19 common peaks were found, which came from S. baicalensis except that two peaks came from S. flavescens, and eight peaks were baicalin/trifolirhizin, wogonoside, luteolin, baicalein, oroxylin A, 5, 8-dihydroxy-6, 7-dimethoxyflavone, wogonin and formononetin/chrysin respectively. Under the detective wavelength 365 nm, 22 common peaks were found, which thirteen peaks came from S. baicalensis and six peaks came from S. flavescens. Among them, eleven peaks were baicalin, wogonoside, baicalein, oroxylin A, 5,8-dihydroxy-6, 7-dimethoxyflavone, wogonin, chrysin/7-methoxyflavone, 2', 4-dihydroxy-4', 6'-dimethoxychalcone, 4, 4'-dimethoxychalcone, xanthohumol and kuraridin respectively.. The common peaks of HPLC fingerprint of the flavonoids of Kushen Tang, which were obtained under the two detective wavelengths 280 nm and 365 nm, came from S. baicalensis and S. flavescens. It was more scientific and comprehensive that establishing the HPLC fingerprint of the flavonoids of Kushen Tang under the detective wavelength 365 nm.

Topics: Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Plant Roots; Plants, Medicinal; Rehmannia; Scutellaria baicalensis; Sophora

To compare and analyze the effects of the contents of the active components in Radix Scutellariae by different processing and extracting methods.. The raw and processed Radix Scutellariae were used, and the concentration of baicalin, wogonoside, baicalein, wogonin were determined by HPLC after five extracting techniques. HPLC method was performed on methanol-acetonitrile-water-acetic acid (18: 25: 57: 0.2) as the mobile phase at 30 C; The chromatographic column was Agilent Zorbax Extend-C18 (4.6 mm x 250 mm, 5 microm); The flow rate was 0.8 mL min(-1), and the detection wavelength was 275 nm.. the ratios of baicalein and wogonin in raw material were 5.41%, 1.30% respectively by enzymatic extraction, which is higher than other extracting methods by raw material; The ratios of baicalin and wogonoside in raw material were 10.11% and 3.55% by ethanol of 60%, which is higher than other extracting methods; Different extracting methods have no evident effects on processed materials, enzymatic extraction is the best. The ratio of baicalin, wogonoside and baicalein, wogonin is 10.63%, 3.60% and 1.54%, 0.59%.. Different methods have evident extracting effects on the four active components with raw material, but have no evident effects with processed material. According to different active components most suitable extracting method should be adopted.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Glucosides; Hot Temperature; Plant Roots; Plants, Medicinal; Reproducibility of Results; Scutellaria baicalensis; Technology, Pharmaceutical

The phytochemicals betulinic acid (1), wogonin (2) and oroxindin (3) isolated from the aerial parts of Bacopa monnieri and Holmskioldia sanguinea showed significant antifungal activity against the two fungi Alternaria alternata and Fusarium fusiformis. Inhibition of root growth germination of wheat seeds was observed for all three compounds which showed 100% inhibition at 10 micro g/mL. Compounds (1) and (2) showed potent inhibition of Alternaria alternata compared with oroxindin at a concentration of 4 micro g/mL, whereas compound (2) was an effective inhibitor of both fungi. The structures of the compounds were established by spectral and chemical studies.

Topics: Antifungal Agents; Bacopa; Betulinic Acid; Chromones; Flavanones; Glucuronates; Humans; Microbial Sensitivity Tests; Mitosporic Fungi; Pentacyclic Triterpenes; Phytotherapy; Plant Extracts; Plant Roots; Seeds; Triterpenes; Verbenaceae

The four flavonoids baicalin, baicalein, wogonoside, and wogonin have been isolated from Scutellaria baicalensis (Lamiaceae) root extract as main bioactive components. We evaluated their antimutagenic activity as potential chemopreventive natural substances using the Ames test. Free radical scavenging properties have also been measured with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Both crude extract and isolated flavone glycosides and aglycones have antimutagenic and free radical scavenging properties. The lipophilic flavone baicalein has been confirmed to be the most powerful antimutagen among the compounds tested.

Topics: Antimutagenic Agents; Antioxidants; Biphenyl Compounds; Flavanones; Flavonoids; Free Radical Scavengers; Glucosides; Hydrazines; Mutagenicity Tests; Picrates; Plant Extracts; Plant Roots; Scutellaria baicalensis

Wogonin (WG), wogonoside (WGS), and 3,5,7,2',6'-pentahydroxyl flavanone (PHF) were isolated from Scutellaria baicalensis, and their effects on histamine, leukotriene B(4) (LTB(4)), and immunoglobulin E (IgE) were examined in rats, observing for a manifestation of a type I allergic reaction. WG and WGS in the amounts of 10 and 100 microM were shown to markedly inhibit histamine release in cells stimulated with calcium ionophore A23187 or compound 48/80. PHF exerted inhibitory activity only at 100 microM. In the case of LTB(4), WG, WGS and PHF markedly inhibited LTB(4) production at the concentration of 100 microM. We also find that the increase in the IgE content induced by concanavalin A (ConA) was alleviated in the presence WG and WGS, while the inhibitory effect of PHF was much weaker. However, the magnitude of inhibitory effect observed on the content of lipid peroxidation induced by ConA was in order of PHF > WG > WGS, with PHF being the strongest. Interestingly, WG and WGS with the methoxyl group strongly inhibited histamine and IgE production, whereas PHF with the hydroxyl group in the B ring was much stronger than WG and WGS against lipid peroxidation. Based on data, it was concluded that the flavonoid components, WG, WGS, and PHF, may block a common pathway for the release of histamine and LTB(4), and that the IgE level is responsible for the lipid peroxidation induced by ConA.

Topics: Animals; Ascitic Fluid; Flavanones; Flavonoids; Glucosides; Histamine; Immunoglobulin E; Leukotriene B4; Lipid Peroxidation; Lipid Peroxides; Lymph Nodes; Lymphocytes; Mesentery; Molecular Structure; Rats; Scutellaria baicalensis