wogonin and baicalin

Ischemia is a common pathological condition present in many neurodegenerative diseases, including ischemic stroke, retinal vascular occlusion, diabetic retinopathy, and glaucoma, threatening the sight and lives of millions of people globally. Ischemia can trigger excessive oxidative stress, inflammation, and vascular dysfunction, leading to the disruption of tissue homeostasis and, ultimately, cell death. Current therapies are very limited and have a narrow time window for effective treatment. Thus, there is an urgent need to develop more effective therapeutic options for ischemia-induced neural injuries. With emerging reports on the pharmacological properties of natural flavonoids, these compounds present potent antioxidative, anti-inflammatory, and antiapoptotic agents for the treatment of ischemic insults. Three major active flavonoids, baicalein, baicalin, and wogonin, have been extracted from

Topics: Brain; Flavanones; Flavonoids; Humans; Oxidative Stress; Retina

Ocular diseases such as cataract, glaucoma, age-related macular degeneration (AMD), and diabetic retinopathy are the leading causes of blindness. The elderly population is at particular risk of developing one or more of these age-related ocular diseases. By virtue of multiple bioactivities, effort has been made to develop dietary flavonoids as complimentary therapies for ocular disorders. Dietary intake of flavonoids has been reported to reduce the risk of cataract and AMD. This review focuses on the main flavones baicalein, baicalin, and wogonin isolated from the Chinese medicinal herb, Scutellariae radix (SR), which has been widely used in Asian countries for the treatment of many diseases. Interest in SR has grown recently following new findings that suggest multiple routes of therapeutic action. This review will summarize the diverse pharmacological properties, therapeutic roles, and mechanisms of these flavones of SR in cellular and animal models of ocular diseases.

Topics: Animals; Drugs, Chinese Herbal; Eye Diseases; Flavanones; Flavonoids; Humans; Molecular Structure; Scutellaria baicalensis

Traditional Chinese medicines have been recently recognized as a new source of anticancer drugs and new chemotherapy adjuvant to enhance the efficacy of chemotherapy and to ameliorate the side effects of cancer chemotherapies however their healing mechanisms are still largely unknown. Scutellaria baicalensis is one of the most popular and multi-purpose herb used in China traditionally for treatment of inflammation, hypertension, cardiovascular diseases, and bacterial and viral infections. Accumulating evidence demonstrate that Scutellaria also possesses potent anticancer activities. The bioactive components of Scutellaria have been confirmed to be flavones. The major constituents of Scutellaria baicalensis are Wogonin, Baicalein and Baicalin. These phytochemicals are not only cytostatic but also cytotoxic to various human tumor cell lines in vitro and inhibit tumor growth in vivo. Most importantly, they show almost no or minor toxicity to normal epithelial and normal peripheral blood and myeloid cells. The antitumor functions of these flavones are largely due to their abilities to scavenge oxidative radicals, to attenuate NF-kappaB activity, to inhibit several genes important for regulation of the cell cycle, to suppress COX-2 gene expression and to prevent viral infections. The tumor-selectivity of Wogonin has recently been demonstrated to be due to its ability to differentially modulate the oxidation-reduction status of malignant vs. normal lymphocytic cells and to preferentially induce phospholipase C gamma 1, a key enzyme involved in Ca(2+) signaling, through H(2)O(2) signaling in malignant lymphocytes. This review is aimed to summarize the research results obtained since the last 20 years and to highlight the recently discovered molecular mechanisms.

Topics: Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Glucuronidase; Humans; Neoplasms; Phytotherapy; Plant Extracts; Scutellaria baicalensis; Topoisomerase II Inhibitors

Topics: Animals; Chitosan; Drugs, Chinese Herbal; Flavanones; Flavonoids; Hydrogels; Mouth Mucosa; Periodontal Diseases; Plant Extracts; Plant Roots; Scutellaria baicalensis; Swine

Motivated by the wide use of Scutellariae Radix (SR) in the food and pharmaceutical industries, a rapid and non-destructive near-infrared spectroscopy (NIRS) method was developed for the simultaneous analysis of three main active components in raw SR and SR processed by stir-frying with wine. From seven geographical areas, 58 samples were collected. The reference contents for the SR components baicalin, baicalein, and wogonin were determined by high-performance liquid chromatography. Two multivariate analysis methods, partial least-squares (PLS) regression as a linear regression method and artificial neural networks (ANN) as a nonlinear regression method, were applied to the NIR data, and their results were compared. In the PLS model, different model parameters (i.e., 11 spectral pre-treatment methods), spectral region, and latent variables were investigated to optimize the calibration model; additionally, the ANN model was applied with five different spectral pre-treatment methods and six algorithms. For the optimal model parameters, the correlation coefficients of the calibration set for baicalin, baicalein, and wogonin were 0.9979, 0.9786, and 0.9773, respectively; the correlation coefficients of the prediction set were 0.9756, 0.9843, and 0.9592, respectively; the root mean square error of validation values were 0.215, 0.321, and 0.174, respectively. The optimal NIR models were then employed to analyze the effects of processing and geographical regions on analyte contents. The established NIR methods were robust, accurate, and reproducible. NIRS may be a promising approach for the routine screening and quality control of traditional Chinese medicines.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Geography; Least-Squares Analysis; Medicine, Chinese Traditional; Multivariate Analysis; Plant Roots; Scutellaria baicalensis; Spectroscopy, Near-Infrared

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death in the world. Inhibitor of differentiation 1 (Id1) is overexpressed in NSCLC and involved in promoting its progression and metastasis. Identifying natural compounds targeting Id1 may have utility in NSCLC treatment. Here, we sought to determine whether the anti-tumor activities of

Topics: A549 Cells; alpha7 Nicotinic Acetylcholine Receptor; Animals; Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Flavanones; Flavonoids; Guanosine Triphosphate; Humans; Inhibitor of Differentiation Protein 1; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis; Phenotype; Phosphorylation; Plant Extracts; Scutellaria; Shelterin Complex; Telomere-Binding Proteins

Rhubarb-Radix scutellariae is a classic herb pair, which is commonly used to clear away heat and toxin in clinic. The aim of this study was to investigate the influence of compatibility of Rhubarb and Radix scutellariae on the pharmacokinetic behaviors of anthraquinones and flavonoids in rat plasma.. Eighteen rats were randomly divided into three groups, and were orally administered Rhubarb and/or Radix scutellariae extracts. A sensitive and rapid UPLC-MS/MS method was developed and validated to determine the concentrations of baicalin, baicalein, wogonside, wogonin, rhein, and emodin in rat plasma. The concentrations of phase II conjugates of flavonoid aglycones and anthraquinone aglycones were also determined after hydrolyzing the plasma with sulfatase.. Compared with administration of Radix scutellariae alone, co-administration of Rhubarb significantly decreased the first maximum plasma concentration (C. Compatibility of Rhubarb and Radix scutellariae can significantly affect the pharmacokinetic behaviors of characteristic constituents of the two herbs. The cause of these pharmacokinetic differences was further discussed combined with the in vivo ADME (absorption, disposition, metabolism, and excretion) processes of anthraquinones and flavonoids.

Topics: Animals; Anthraquinones; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Herb-Drug Interactions; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Rheum; Scutellaria baicalensis; Tandem Mass Spectrometry

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

As an important herbaceous plant, Scutellaria baicalensis Georgi (Chinese skullcap) is geographically widespread and commonly used throughout the world. In the Chinese medicine market, S. baicalensis has been divided into two primary types, "Ku Qin" (WXR) and "Tiao Qin" (TST). Moreover, TST is also divided into different grades according to the diameter of roots. To explore the distribution patterns of the contents of five biologically activate ingredients (FBAI), we used six-year-old cultivated S. baicalensis and analyzed its growth characteristics as well as the quality difference among different types and diameters in roots. Throughout the entire root, we discovered that contents of the FBAI all initially increased and subsequently decreased from the top to the bottom of the roots. The baicalin content of WXR was less than that of TST. On the contrary, the contents of baicalein, wogonin, and oroxylin A in WXR were up to about two times higher than that in TST. We also found that the 0 to 40 cm part of the S. baicalensis root possessed about 87% of the root biomass and about 92% of the contents of the active ingredients.

Topics: Drugs, Chinese Herbal; Flavanones; Flavonoids; Plant Roots; Scutellaria baicalensis

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

Wogonin and baicalein are bioactive flavones in the popular Chinese herbal remedy Huang-Qin (Scutellaria baicalensis Georgi). These specialized flavones lack a 4'-hydroxyl group on the B ring (4'-deoxyflavones) and induce apoptosis in a wide spectrum of human tumor cells in vitro and inhibit tumor growth in vivo in different mouse tumor models. Root-specific flavones (RSFs) from Scutellaria have a variety of reported additional beneficial effects including antioxidant and antiviral properties. We describe the characterization of a new pathway for the synthesis of these compounds, in which pinocembrin (a 4'-deoxyflavanone) serves as a key intermediate. Although two genes encoding flavone synthase II (FNSII) are expressed in the roots of S. baicalensis, FNSII-1 has broad specificity for flavanones as substrates, whereas FNSII-2 is specific for pinocembrin. FNSII-2 is responsible for the synthesis of 4'-deoxyRSFs, such as chrysin and wogonin, wogonoside, baicalein, and baicalin, which are synthesized from chrysin. A gene encoding a cinnamic acid-specific coenzyme A ligase (SbCLL-7), which is highly expressed in roots, is required for the synthesis of RSFs by FNSII-2, as demonstrated by gene silencing. A specific isoform of chalcone synthase (SbCHS-2) that is highly expressed in roots producing RSFs is also required for the synthesis of chrysin. Our studies reveal a recently evolved pathway for biosynthesis of specific, bioactive 4'-deoxyflavones in the roots of S. baicalensis.

Topics: Acyltransferases; Animals; Antioxidants; Apoptosis; Biosynthetic Pathways; Cell Line, Tumor; Cytochrome P-450 Enzyme System; Flavanones; Flavones; Flavonoids; Gene Expression Regulation, Plant; Humans; Mice; Plant Extracts; Plants, Medicinal; Scutellaria baicalensis

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

Here, three structurally related polyphenols found in the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, were examined for its effects on inflammatory responses by monitoring the effects of baicalin, baicalein, and wogonin on lipopolysaccharide (LPS)-mediated vascular inflammatory responses. We found that each compound inhibited LPS-induced barrier disruption, expression of cell adhesion molecules (CAMs), and adhesion/transendothelial migration of monocytes to human endothelial cells. Each compound induced potent inhibition of phorbol-12-myristate 13-acetate and LPS-induced endothelial cell protein C receptor shedding. It also suppressed LPS-induced hyperpermeability and leukocytes migration in vivo. Furthermore, each compound suppressed the production of tumor necrosis factor-α or interleukin-6 and the activation of nuclear factor-κB or extracellular regulated kinases 1/2 by LPS. Moreover, treatment with each compound resulted in reduced LPS-induced lethal endotoxemia. These results suggest that baicalin, baicalein, and wogonin posses anti-inflammatory functions by inhibiting hyperpermeability, expression of CAMs, and adhesion and migration of leukocytes, thereby endorsing its usefulness as a therapy for vascular inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents; Dose-Response Relationship, Drug; Flavanones; Flavonoids; Human Umbilical Vein Endothelial Cells; Humans; Inflammation Mediators; Male; Mice; Mice, Inbred C57BL

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

It is well known that the expression level of secretory group IIA phospholipase A2 (sPLA2-IIA) is elevated in inflammatory diseases and lipopolysaccharide (LPS) up-regulates the expression of sPLA2-IIA in human umbilical vein endothelial cells (HUVECs). Here, three structurally related polyphenols found in the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, were examined for its effects on the expression and activity of sPLA2-IIA in HUVECs and mouse. Prior treatment of cells or mouse with baicalin, baicalein, and wogonin inhibited LPS-induced expression and activity of sPLA2-IIA. And each compound suppressed the activation of cytosolic phospholipase A2 (cPLA2) and extracellular signal-regulated kinase (ERK) 1/2 by LPS. Therefore, these results suggest that baicalin, baicalein, and wogonin inhibited LPS mediated expression of sPLA2-IIA by suppression of cPLA2 and ERK 1/2.

Topics: Animals; Drugs, Chinese Herbal; Flavanones; Flavonoids; Gene Expression Regulation, Enzymologic; Group II Phospholipases A2; Human Umbilical Vein Endothelial Cells; Humans; Lipopolysaccharides; Male; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Phospholipases A2, Cytosolic

Vascular inflammatory process has been suggested to play a key role in initiation and progression of atherosclerosis, a major complication of diabetes mellitus. Thus, in this study, we attempted to determine whether three structurally related polyphenols found in the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, can suppress vascular inflammatory processes induced by high glucose (HG) in human umbilical vein endothelial cells (HUVECs) and mice. Data showed that HG induced markedly increased vascular permeability, monocyte adhesion, expressions of cell adhesion molecules (CAMs), formation of reactive oxygen species (ROS) and activation of nuclear factor (NF)-κB. Remarkably, all of the above mentioned vascular inflammatory effects of HG were attenuated by pretreatment with baicalin, baicalein, and wogonin. Vascular inflammatory responses induced by HG are critical events underlying development of various diabetic complications, therefore, our results suggest that baicalin, baicalein, and wogonin may have significant therapeutic benefits against diabetic complications and atherosclerosis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Astragalus propinquus; Cell Adhesion Molecules; Diabetes Complications; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucose; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Mice; NF-kappa B; Reactive Oxygen Species; Vasculitis

To establish an HPLC method for the content determination of baicalin, wogonin, chlorogenic acid, caffeic acid, cichoric acid, corynoline and adenosine in Pudilan Xiaoyan oral liquid.. The analysis was performed on a Phenomenex Luna C18 column (4.6 mm x 250 mm, 5 µm) with a gradient mobile phase of methanol-0.1% trifluoroacetic acid solution system at flow rate of 1.0 mL · min(-1). The detective wavelength was at 280 nm. The column temperature was 30 °C.. The standard curves of seven studied components show good linearity in their concentration ranges with r ≥ 0.999 6. The average recovery was 98.73%-102.1% with RSD less than 2.6%.. The method is rapid, simple and accurate, and can be applied for the quality control of Pudilan Xiaoyan oral liquid.

Topics: Caffeic Acids; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Succinates

To observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.. RP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).. HJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Topics: Animals; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucose; Iridoids; Models, Animal; Neurons; Oxygen; Rats; Reperfusion Injury

The objective of the present study was to investigate the therapeutic efficacy of flavonoid components in Scutellaria baicalensis on proliferation, metastasis and lung cancer-associated inflammation during nicotine induction in the A549 and H1299 lung cancer cell lines. After experimental period, augmentation of proliferation was observed, accompanied by marked decrease in apoptotic cells in nicotine-induced lung cancer cells; additionally, nicotine-exposed cells exhibited increased invasive and migratory abilities based on invasion and wound-healing assay. Flavones in Scutellaria, baicalin, baicalein and wogonin significantly counteracted the above deleterious changes. Moreover, assessment of tumor apoptotic and metastatic factors on mRNA levels by quantitative PCR and protein levels by western blotting revealed that these phytochemical treatments effectively negated nicotine-induced upregulated expression of bcl-2, bcl-2/bax ratio, caspase-3, matrix metalloproteinase (MMP)-2 and MMP-9 as well as downregulated expression of bax. Further analysis of inflammatory markers such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 in cell culture supernatant and mRNA and protein expression of nuclear transcription factor-kappaB (NF-κB) and I kappa B-alpha (IκB-α) was carried out to substantiate the anti-inflammatory effect of flavones in Scutellaria in nicotine-exposed lung cancer cells. The therapeutic effects observed in the present study are attributed to the potent potential against proliferation, metastasis and inflammatory microenvironment by flavonoid components in Scutellaria in nicotine-induced lung cancer cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Line, Tumor; Cell Proliferation; Flavanones; Flavonoids; Gene Expression Regulation, Neoplastic; Humans; In Vitro Techniques; Inflammation; Lung Neoplasms; Neoplasm Metastasis; Nicotine; Plant Extracts; 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

Inhibition of high mobility group box 1 (HMGB1) protein and restoration of endothelial integrity is emerging as an attractive therapeutic strategy in the management of sepsis. Here, three structurally related polyphenols found in the Chinese herb Huang Qui, baicalin (BCL), baicalein (BCN), and wogonin (WGN), were examined for their effects on lipopolysaccharide (LPS)- or cecal ligation and puncture (CLP)-mediated release of HMGB1 and on modulation of HMGB1-mediated inflammatory responses. According to our data, BCL, BCN, and WGN inhibited the release of HMGB1 and down-regulated HMGB1-dependent inflammatory responses in human endothelial cells. BCL, BCN, and WGN also inhibited HMGB1-mediated hyperpermeability and leukocyte migration in mice. In addition, treatment with BCL, BCN, and WGN reduced CLP-induced release of HMGB1 and sepsis-related mortality and pulmonary injury in mice. These results indicate that BCL, BCN, and WGN could be candidate therapeutic agents for various severe vascular inflammatory diseases owing to their inhibition of the HMGB1 signaling pathway.

Topics: Animals; Capillary Permeability; Cell Movement; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Flavanones; Flavonoids; HMGB1 Protein; Human Umbilical Vein Endothelial Cells; Humans; Male; Mice; Mice, Inbred C57BL; Protective Agents

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

Solute carrier transporters (SLCs), in particular the organic anion transporters (OATs), OAT polypeptides (OATPs), and organic cation transporters (OCTs/OCTNs), are the important membrane proteins responsible for the cellular influx of various drugs. Baicalein (BA), baicalin (BG), and wogonin (WG) are the three major bioactive components of Scutellaria baicalensis. In this study, we evaluated the inhibitory effects of BA, BG, and WG on the cellular uptake of specific substrates mediated by the essential SLCs in human embryonic kidney-293 cells. Our data demonstrated that BA and WG significantly inhibit the OAT1-, OAT3-, and OATP1B3-mediated uptake; BG effectively reduces the influx of substrates of OAT3, OAT4, OATP1B3, and OATP2B1; WG is a potent inhibitor of OCT3. Our further kinetic analysis derived the IC50 values of these compounds with pronounced inhibitory effects on SLCs, particularly the inhibitions of WG on OAT1 and OCT3 and that of BA and WG on OAT3. Our study comprehensively evaluated the inhibitory effects of three bioactive components of Scutellaria baicalensis on the uptake of specific substrates mediated by the essential SLC transporters, which suggested that precautions will be needed when coadministrating drugs with Scutellaria baicalensis so as to prevent the unfavorable drug-drug/herb interactions in human.

Topics: Biological Transport, Active; Cell Line; Flavanones; Flavonoids; Humans; Organic Anion Transport Protein 1; Organic Anion Transporters, Sodium-Independent; Plant Extracts; Scutellaria baicalensis; Solute Carrier Organic Anion Transporter Family Member 1B3

This study was performed to develop methods for the chromatographic determination of biomarkers in Hwangryunhaedok-tang (HHT) and the quantitative evaluation of commercial HHT. To develop an analytical method, an RP-amide column (2.7 μm, 4.6 × 100 mm, Halo: Supelco, Bellefonte, PA) was used with a gradient solvent system of mixed acetonitrile and 0.1 % phosphoric acid/water and an ultra performance liquid chromatography-diode array detector. The method was validated by specificity, linearity, accuracy (recovery) and precision tests (repeatability, intra and inter-day). The correlation coefficients (R (2)) of biomarkers were calculated as 0.9998-1.000 and their ranges were as follows: geniposide (62.5-1,000.0 μg/ml), berberine (31.3-500.0 mg/ml), palmatine (31.3-500.0 μg/ml), baicalin (125.0-1,500.0 μg/ml), baicalein (15.6-250.0 μg/ml) and wogonin (5.2-125.0 μg/ml), respectively. The limit of detection was 0.34-4.01 μg/ml, and the limit of quantification was 1.02-12.16 μg/ml. The intra-day and inter-day precision of six components were revealed as 0.02-2.48 % as a relative standard deviation (RSD). The repeatability value of biomarkers in three different concentrations of HHT was 0.29-2.98 % (RSD value) and recovery was 95.72-104.90 %. Among several extraction methods tested, biomarker content was higher with the 20 times extraction (20TE) and mixture of extract powder (MEP) methods than with any other method, and some differences among diverse pharmaceutical medicines were revealed. The validation data indicated that the method developed is suited to the determination of six marker compounds in HHT. The content of biomarkers by simultaneous analysis was evaluated in 20TE, MEP, USA formula and Taiwan formula.

Topics: Berberine; Drugs, Chinese Herbal; Flavanones; Flavonoids; Iridoids

Scutellaria baicalensis extract (SbE) has been shown to exert chemopreventive effects on several types of cancer. Baicalin, a hydrophilic flavonoid found in SbE, may have opposing effects that decrease the antitumor potential of SbE against colorectal cancer. In this study, after removing baicalin, we prepared an aglycone-rich fraction (ARF) of SbE and evaluated its anti-proliferative activity and mechanisms of action. The flavonoids found in ARF, baicalin fraction (BF) and SbE were determined by high-performance liquid chromatography (HPLC). The effects of ARF, BF, SbE and representative flavonoids on the proliferation of HCT-116 and HT-29 human colorectal cancer cells were determined by an MTS assay. The cell cycle, the expression of cyclins A and B1 and cell apoptosis were assayed using flow cytometry. Apoptosis-related gene expression was visualized by quantitative real-time polymerase chain reaction (PCR), and mitochondrial membrane potential was estimated following staining with JC-1. HPLC analysis showed that ARF contained two hydrophobic flavonoids, baicalein and wogonin, and that BF contained only baicalin. SbE had little anti-proliferative effect on the colorectal cancer cells; cancer cell growth was even observed at certain concentrations. ARF exerted potent anti-proliferative effects on the cancer cells. By contrast, BF increased cancer cell growth. ARF arrested cells in the S and G2/M phases, increased the expression of cyclins A and B1, and significantly induced cell apoptosis. Multiple genes in the mitochondrial pathway are involved in ARF-induced apoptosis, and subsequent cellular functional analysis validated the involvement of this pathway. These results suggest that removing baicalin from SbE produces an ARF that significantly inhibits the growth of colorectal cancer cells, and that the mitochondrial apoptotic pathway plays a role in hydrophobic flavonoid-induced apoptosis.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Line, Tumor; Cell Proliferation; Chemoprevention; Colorectal Neoplasms; Cyclin A; Cyclin B1; Enzyme Inhibitors; Flavanones; Flavonoids; G2 Phase Cell Cycle Checkpoints; Gene Expression; Humans; Iridoids; Membrane Potential, Mitochondrial; Mitochondria; Phytotherapy; Plant Extracts; S Phase Cell Cycle Checkpoints; Scutellaria baicalensis

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

Flavonoid reference standards were targeted-prepared from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis. With HPLC-MS analysis of Scutellariae Radix, 19 flavonoid components were identified by analyzing and comparing their retention times, ultraviolet spectra, and mass spectrometry data with literature. The separation and purification protocols of all targeted flavonoid reference standards were optimally designed according to the results of HPLC-MS analysis and related literature. The ethanol extract of Scutellariae Radix was suspended in water and extracted with petroleum ether, ethyl acetate, and n-butanol successively. The ethyl acetate extract and n-butanol extract were separately subjected to primary separation by low pressure reverse phase preparative chromatography. Then the fractions containing targeted compounds were further purified by low pressure reverse and normal phases preparative chromatography. Finally, baicalin and wogonoside reference standards were obtained from n-butanol extract; baicaelin, wogonin, and oroxylin A reference standards were obtained from ethyl acetate extract. The structures of the 5 reference standards were identified by mass spectrometry (MS) and 1H nuclear magnetic resonance (1H NMR) spectroscopy. The HPLC analytical results showed that the purities of the 5 reference standards were all above 98%. It is demonstrated that the rapid targeted-preparation method under the guidance of the HPLC-MS analysis is applicable for the isolation and preparation of chemical components in traditional Chinese medicines.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Mass Spectrometry; Reference Standards; Scutellaria baicalensis

Scutellariae Radix (root of Scutellaria baicalensis, SR) contains numerous flavonoids such as baicalin, baicalein, and wogonin. This study investigated the pharmacokinetics and tissue distribution of flavonoids and their metabolites in rats after repeated dosing of a SR decoction. Sprague-Dawley rats were orally administered SR at 2 g/kg for seven doses. After the 7th dose, blood samples were withdrawn at specific times and organs, including the liver, kidney, lung, and brain, and collected. The concentrations of baicalein and wogonin in the serum and various tissues were assayed by HPLC before and after hydrolysis with glucuronidase and sulfatase. Baicalein and wogonin were not detected in the serum, and the molecules found were their glucuronides/sulfates. In tissues, the free forms of baicalein and wogonin appeared in the liver, kidney, and lung in addition to their glucuronides/sulfates. Baicalein was the major form in the lung, whereas baicalein glucuronides/sulfates were the major forms in the liver and kidney. Wogonin was the major form in the liver, kidney, lung, and traces of wogonin glucuronides/sulfates were detected in the kidney and liver. Neither baicalein and wogonin nor their glucuronides/sulfates were detected in the brain. In conclusion, the glucuronides/sulfates of baicalein and wogonin were exclusively present in the circulation, whereas their free forms appeared in the lung, liver, and kidney.

Topics: Administration, Oral; Animals; Brain; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Kidney; Liver; Lung; Male; Plant Extracts; Plant Roots; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Tandem Mass Spectrometry; Tissue Distribution

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

Based on our previous observation, the whole Scutellaria baicalensis extract (SbE) did not show significant breast cancer cell inhibitory effect. In this study, we isolated a baicalin-deprived-fraction (SbF1) of Scutellaria baicalensis, and baicalin-fraction (SbF3), and evaluated their anti-breast cancer properties using MCF-7 cells. The content of four flavonoids in extract/fractions were determined using high performance liquid chromatography. Analytical data showed that in SbF1, the major constituents are baicalein and wogonin, while SbF3 only contains baicalin. The antiproliferative effects of fractions and SbE were assayed using modified trichrome stain method. SbF1 showed significant antiproliferative effect. Treated with 100mug/ml of SbF1 for 72h inhibited MCF-7 cell growth by 81.6%, while in the same treatment concentration, SbF3 increased cell growth by 22.6%. SbF1 was recognized as an active fraction of SbE. The effects of four flavonoids in SbE, scutellarin, baicalin, baicalein and wogonin, were determined, and data showed that baicalein and wogonin significantly inhibited MCF-7 cell growth. In contrast, in certain concentrations, scutellarin and baicalin increased cancer cell growth. The effects of SbF1 on cell cycle and apoptosis were assayed using flow cytometry. SbF1 arrested MCF-7 cells in S- and G2/M-phases, and significantly increased induction of cell apoptosis. These combined phytochemical and biological data provide evidence for further chemopreventive studies of the baicalin-deprived SbE on breast cancer.

Topics: Antineoplastic Agents, Phytogenic; Apigenin; Apoptosis; Breast Neoplasms; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Female; Flavanones; Flavonoids; Glucuronates; Humans; Phytotherapy; Plant Extracts; Scutellaria baicalensis

An on-line method, based on coupling dynamic ultrasonic extraction (DUE), continuously sampling the suspension of sample and solvent, high performance liquid chromatographic separation with diode array detection, has been developed for the determination of the flavonoids, including baicalin, baicalein and wogonin, from the root of Scutellaria baicalensis Georgi. Variables influencing the DUE were evaluated by orthogonal test. The extraction yields of baicalin, baicalein and wogonin in the roots of S. baicalensis Georgi obtained from five different cultivated areas are 73.8-131.5 microg mg(-1) (RSD< or =6.24%), 6.8-15.9 microg mg(-1) (RSD< or =5.36%) and 4.4-14.3 microg mg(-1) (RSD< or =5.30%), respectively. The limits of detection for baicalin, baicalein and wogonin are 0.30, 0.37 and 0.41 microg mL(-1), respectively. Linearity is from 0.55 to 109 microg mL(-1) for baicalin, from 0.51 to 105 microg mL(-1) for baicalein and from 0.53 to 102 microg mL(-1) for wogonin. Compared with off-line continuous flow-DUE, the proposed method would be more convenient for the determination of the analytes and the rapid optimization of the extraction process. The extraction yields of flavonoids obtained by the proposed method are comparable with those obtained by dynamic microwave assisted extraction, static ultrasonic extraction and reflux extraction. The result indicated that the proposed method is suitable to determine the active components in Chinese herbal medicine.

Topics: Chromatography, High Pressure Liquid; Evaluation Studies as Topic; Flavanones; Flavonoids; Limit of Detection; Online Systems; Particle Size; Plant Roots; Reference Standards; Regression Analysis; Rheology; Scutellaria baicalensis; Solvents; Ultrasonics

A sensitive, rapid and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed and validated for the simultaneous determination of baicalin, baicalein, wogonin, berberine, palmatine and jatrorrhizine in scutellaria-coptis herb couple in rat plasma. After protein precipitation with acetonitrile and 0.1% NaH(2)PO(4), chromatography was performed using a C(18) column, with gradient elution with 0.1% formic acid and acetonitrile at 0.25 ml/min. All analytes including internal standards were monitored under positive ionization conditions by selected reaction monitoring with an electrospray ionization source. The lower limit of quantification was 10 ng/ml for baicalin, baicalein and wogonin, and 0.6 ng/ml for berberine, palmatine and jatrorrhizine. The validated method was applied in pharmacokinetic studies after oral administration of Yiqing Capsule and Gegen-Qinlian Tablet to rats.

Topics: Administration, Oral; Animals; Berberine; Berberine Alkaloids; Chromatography, Liquid; Coptis; Flavanones; Flavonoids; Male; Medicine, Chinese Traditional; Rats; Rats, Sprague-Dawley; Scutellaria; Tandem Mass Spectrometry

Scutellaria baicalensis root is widely used in China as an adjuvant to orthodox chemotherapy of lung cancer. However, functional biomarkers of this plant for anti-lung cancer activity have not yet been reported. We therefore determined the growth inhibition activity by MTT assay of eight solvent extracts of S. baicalensis in the human lung cancer cell line SK-MES-1. This activity was then mapped onto the secondary metabolite profile of crude extracts by principal components analysis (PCA) of proton NMR and HPLC-UV data. NMR- and HPLC-PCA maps revealed highest inhibitory activity for the non-aqueous extracts. The first two components of both maps discriminated extract activity mainly based on the differential content of three compounds, which were then tested individually. The IC(50) values for baicalin (IC(50): 64+/-5 microM), baicalein (IC(50): 80+/-6 microM) and wogonin (IC(50): 39+/-10 microM) were comparable to that of the antineoplastic cisplatin (IC(50): 79+/-16 microM). A partial least squares regression (PLS)-NMR model highly correlated with the corresponding PLS-HPLC model for prediction of inhibition. Secondary metabolite mapping of lung cancer growth inhibitors in crude extracts may be an important first step to qualify Chinese herbal prescriptions required for meaningful clinical trials of such integrated therapies.

Topics: Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Humans; Least-Squares Analysis; Lung Neoplasms; Magnetic Resonance Spectroscopy; Plant Extracts; Principal Component Analysis; Scutellaria

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

The involvement of genes in flavones biosynthesis was investigated in different organs and suspension cells obtained from Scutellaria baicalensis. Three full-length cDNAs encoding phenylalanine ammonia-lyase isoforms (SbPAL1, SbPAL2, and SbAPL3) and one gene encoding cinnamate 4-hydroxylase (SbC4H) from S. baicalensis were isolated using rapid amplification of cDNA ends (RACE)-PCR. These cDNAs were used together with previously-isolated clones for 4-coumaroyl CoA ligase (4CL) and chalcone synthase (CHS) to show the expression level in different organs of S. baicalensis. These genes were upregulated in suspension cells of S. baicalensis with biotic/abiotic stress factors. The baicalin and baicalein contents in roots were 22 and 107 times higher than those in flowers, respectively. The treatment of suspension cells with methyl jasmonate (MeJa) enhanced the major flavones in S. baicalensis. Cumulatively, the results of this study should advance ability to biosynthesize important and useful medicinal compounds from a variety of plant species.

Topics: Amino Acid Sequence; Chromatography, High Pressure Liquid; Cloning, Molecular; Flavanones; Flavones; Flavonoids; Gene Expression Regulation, Plant; Genes, Plant; Molecular Sequence Data; Organ Specificity; Phenylalanine Ammonia-Lyase; Scutellaria baicalensis; Sequence Alignment; Sequence Analysis, DNA; Time Factors; Trans-Cinnamate 4-Monooxygenase

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 do some comparative study on chemical composition between the cultivated and wild samples of Radix Scutellariae.. Thirty three samples of Radix Scutellariae were collected from different habitats. An HPLC method for fingerprint identification and simultaneous determination of baicalin, baicalein and wogonin was established, and the content of ethanolic extractive was measured according to the method in Chinese Pharmacopoeia.. The number of chromatographic peaks in fingerprint of wild sample was more than that of cultivated sample in Hebei and Inner Mongolia. In 95% confidence intervals, the contents of baicalin, baicalein and wogonin were (15.89 +/- 1.52)%, (1.04 +/- 0.26)% and (0.27 +/- 0.07)%, respectively in the cultivated samples and (11.93 +/- 1.62)%, (1.03 +/- 0.26)% and (0.27 +/- 0.06)%, respectively in wild samples. The content of ethanolic extractive was 52.07 +/- 3.05% in the cultivated and (41.21 +/- 2.86)% in the wild within 95% confidence interval.. Slight difference between the fingerprint of the cultivated and wild samples was found, and the contents of baicalin and ethanolic extraction in the cultivated were higher than those in the wild.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Reproducibility of Results; Scutellaria baicalensis; Sensitivity and Specificity

We observed that treatment of prostate cancer cells for 24 h with wogonin, a naturally occurring monoflavonoid, induced cell death in a dose- and time-dependent manner. Exposure of wogonin to LNCaP cells was associated with increased intracellular levels of p21(Cip-1), p27(Kip-1), p53, and PUMA, oligomerization of Bax, release of cytochrome c from the mitochondria, and activation of caspases. We also confirmed the role of p53 by noting that knock-in in p53 expression by transfecting p53 DNA increased wogonin-induced apoptosis in p53-null PC-3 cells. To study the mechanism of PUMA up-regulation, we determined the activities of PUMA promoter in the wogonin treated and untreated cells. Increase of the intracellular levels of PUMA protein was due to increase in transcriptional activity. Data from chromatin immunoprecipitation (ChIP) analyses revealed that wogonin activated the transcription factor p53 binding activity to the PUMA promoter region. We observed that the up-regulation of PUMA mediated wogonin cytotoxicity. Further characterization of the transcriptional response to wogonin in HCT116 human colon cancer cells demonstrated that PUMA induction was p53-dependent; deficiency in either p53 or PUMA significantly protected HCT116 cells against wogonin-induced apoptosis. Also, wogonin promoted mitochondrial translocation and multimerization of Bax. Interestingly, wogonin (100 microM) treatment did not affect the viability of normal human prostate epithelial cells (PrEC). Taken together, these results indicate that p53-dependent transcriptional induction of PUMA and oligomerization of Bax play important roles in the sensitivity of cancer cells to apoptosis induced by caspase activation through wogonin.

Topics: Apoptosis; Apoptosis Regulatory Proteins; bcl-2-Associated X Protein; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Survival; Cells, Cultured; Cytochromes c; DNA Fragmentation; Epithelial Cells; Flavanones; Flavonoids; Gene Expression Regulation; Humans; Male; Prostate; Proto-Oncogene Proteins; Tumor Suppressor Protein p53

According to the notification for definition of pharmaceuticals from the Director-General of the Pharmaceutical and Food Safety Bureau, Ministry of Health Labour and Welfare of Japan, the roots of Scutellaria baicalensis (Chinese skullcap) and S. lateriflora (skullcap) are classified as "the raw materials exclusively used as pharmaceuticals", but their aerial parts are classified as "non-pharmaceuticals" so, in principle, there are no health claims for these materials and no descriptions of drug-like dosages or administration directions. Dried root of S. baicalensis is also registered in Japanese Pharmacopoeia XV as scutellaria root. Scutellaria root is considered to have the adverse drug reactions of interstitial pneumonia and drug-induced hepatopathy in kampo medicines (Japanese traditional herbal formulations), and baicalin, its major constituent, is considered to be the cause of the adverse reaction. This study was conducted to evaluate the validity of this borderline between pharmaceuticals and non-pharmaceuticals by analyzing the amounts of four flavonoids, including baicalin, in the roots, stems, and leaves of S. baicalensis and S. lateriflora, and in the commercial products herbal tea and dietary supplements prepared from S. lateriflora. These flavonoids were found in the root of S. baicalensis; its aerial parts, however, did not contain them. On the other hand, the amounts of those flavonoids in the aerial parts of S. lateriflora were larger than in the root. Herbal tea and dietary supplements of S. lateriflora obtained commercially also contained those flavonoids, and the dietary supplements contained amounts of them comparable with that in kampo medicine. These results suggest that classification that the aerial parts of S. lateriflora as non-pharmaceuticals in Japan needs reconsideration.

Topics: Dietary Supplements; Flavanones; Flavonoids; Glucuronides; Japan; Plant Extracts; Plant Leaves; Plant Roots; Plant Stems; Scutellaria; Scutellaria baicalensis; Tea

Methanol extract from cultured Scutellaria baicalensis cells inhibited the proliferation of human monocytic leukemia cell line THP-1 and human osteogenic sarcoma cell line HOS. The inhibitory effects of baicalin, baicalein and wogonin, the three major flavonoids contained in the extract, were studied. It should be noted that wogonin did not show the inhibitory effect on human fetal lung normal diploid cell line TIG-1, as compared to the inhibition observed in cancer cells. Physiological analyses in THP-1 cells showed that wogonin induced cell cycle arrest at G(2)/M phase and apoptosis. This is the first report discovering a cancer-specific apoptosis-inducing activity of wogonin.

Topics: Apoptosis; Cell Cycle; Cell Division; Cell Line; Cell Line, Tumor; Cell Proliferation; Diploidy; DNA Fragmentation; Drugs, Chinese Herbal; Flavanones; Flavonoids; G2 Phase; Growth Inhibitors; Humans; Neoplasms; Plant Extracts; Scutellaria baicalensis

Quantitative analysis and chromatographic fingerprinting for the quality evaluation of a Chinese herb Scutellaria baicalensis Georgi using capillary electrophoresis (CE) technique was developed. The separation was performed with a 50.0cm (42.0cm to the detector window)x75mum i.d. fused-silica capillary, and the CE fingerprint condition was optimized using the combination of central composite design and multivariate analysis. The optimized buffer system containing 15mM borate, 40mM phosphate, 15mM SDS, 15% (v/v) acetonitrile and 7.5% (v/v) 2-propanol was employed for the method development, and the baseline separation was achieved within 15min. The determination of the major active components (Baicalin, Baicalein and Wogonin) was carried out using the optimized CE condition. Good linear relationships were provided over the investigated concentration ranges (the values of R(2): 0.9997 for Baicalin, 0.9992 for Baicalein, and 0.9983 for Wogonin, respectively). The average recoveries of these target components ranged between 96.1-105.6%, 98.6-105.2%, and 96.3-105.0%, respectively. CE fingerprints combined with the quantitative analysis can be used for the quality evaluation of S. baicalensis.

Topics: 2-Propanol; Acetonitriles; Buffers; Capillary Electrochromatography; China; Drugs, Chinese Herbal; Flavanones; Flavonoids; Models, Statistical; Molecular Structure; Plant Extracts; Quality Control; Reproducibility of Results; Scutellaria baicalensis; Solvents; Time Factors

The pharmacokinetics of active components such as baicalein, wogonin and oroxylin A were evaluated after oral administration of a purified extract of Scutellaria baicalensis GEORGI (PF-2405) containing the high contents of baicalein, wogonin and oroxylin A to rats. Following oral administration of PF-2405 at 10, 20 and 40 mg/kg dose (equivalent to 4.5, 9.0 and 18 mg/kg baicalein), a major constituent baicalein and its active metabolite baicalin showed dose-linear pharmacokinetics as evidenced by unaltered dose-normalized AUC, dose-normalized Cmax, Ae(0-30h) and GI(30h) values. Following oral administration of PF-2405 at three doses (equivalent to 0.4, 0.8 and 1.6 mg/kg wogonin), dose-normalized Cmax and dose-normalized AUC were comparable between the 20 and 40 mg/kg PF2405 doses, but plasma concentrations of wogonin at 10 mg/kg of PF-2405 were not measurable as they were below limit of quantitation (LOQ; 18 pmol/mL). Following oral administration of PF-2405 at the three doses (equivalent to 1.5, 3.0 and 6.0 mg/kg oroxylin A), the concentrations of oroxylin A in plasma, urine and gastrointestine samples were below the assay LOQ (18 pmol/mL). Significant differences in AUCs, Ae(0-30h) and GI(30h) values for baicalein and baicalin were observed after oral administration of pure baicalein (18 mg/kg) and PF-2405 (40 mg/kg). The increases in AUCs of baicalein and baicalin after oral administration of PF-2405 may have been due to the significant decrease in GO(30h) values for baicalein.

Topics: Administration, Oral; Animals; Chromatography, Liquid; Flavanones; Flavonoids; Gastrointestinal Tract; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Scutellaria baicalensis; Tandem Mass Spectrometry

Scutellaria baicalensis Georgi has been used for the treatment of diverse chronic inflammatory diseases including respiratory disease in oriental medicine and its major components - baicalin, baicalein and wogonin - were reported to have various biological effects. This study investigated whether baicalin, baicalein and wogonin affect basal and ATP-induced mucin release from cultured airway epithelial cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled using (3)H-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of each agent to assess the effects on (3)H-mucin release. The results were as follows: (1) Baicalein did not affect both basal and ATP-induced mucin release significantly. (2) Baicalin and wogonin increased basal mucin release at the highest concentrations (10(-3) m). (3) However, baicalin and wogonin significantly inhibited ATP-induced mucin release. It is concluded that baicalin and wogonin can slightly increase basal mucin release whereas they can inhibit ATP-induced mucin release, by directly acting on airway mucin-secreting cells. It is suggested that baicalin and wogonin be further investigated for the possible use as mucoregulators during the treatment of chronic airway diseases.

Topics: Animals; Anti-Asthmatic Agents; Cells, Cultured; Cricetinae; Drugs, Chinese Herbal; Epithelial Cells; Flavanones; Flavonoids; Male; Mesocricetus; Mucins; Prostaglandin Antagonists; Respiratory Mucosa; 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

Scutellaria baicalensis Georgi (S. baicalensis) is a widely used herb in traditional medical systems of China and Japan. The major constituents of S. baicalensis are flavonoids: baicalein, baicalin, wogonin, which have been associated with various properties, for example: antioxidant, anti-inflammatory, antithrombotic, antibacterial, antiviral. Atherosclerosis is a multiple factors disease, in which the main roles play oxidant stress, endothelium dysfunction and disorders ofhemostasis. It is supposed that flavonoids from S. baicalensis roots in the nearest future could be use in the primary and secondary prevention of atherosclerosis.

Topics: Animals; Anti-Inflammatory Agents; Anticoagulants; Antioxidants; Atherosclerosis; China; Drugs, Chinese Herbal; Flavanones; Flavones; Flavonoids; Free Radical Scavengers; Humans; Japan; Plant Roots; Plants, Medicinal; Scutellaria baicalensis

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

To develop an HPLC-electrochemical detection (HPLC-ECD) method for determination of contents of three effective ingredients baicalin, baicalein and wogonin in Scutellaria baicalensis simultaneously.. 70% ethanol extract of S. baicalensis was separated on a Kromasil C18 column (4.6 mm x 250 mm, 5 microm) using a mixture solution of CH3OH and H2O-THF-H3PO4(80:10:0.2) as mobile phase and detected by Waters2465 electrochemical detector at + 800 mV ( vs. ISAAC). Temperature of column was 30 degrees C.. The linear ranges of baicalin, baicalein and wogonin were 0.045 6-1.14 microg (r = 0.999 6), 0.013-0.325 microg (r = 1) and 0.009 5-0.047 5 microg ( r = 0.999 9), respectively. The average recoveries of baicalin, baicalein and wogonin were 97.9% (n = 5, RSD 2.3%), 97.2% (n = 5, RSD 3.3%) and 103.5% (n = 5, RSD 3.5%), respectively. The contents of three effective ingredients from 14 different germplasm resourses and 9 different growing areas of S. baicalensis were determined, the quality of S. baicalensis was evaluated.

Topics: China; Cluster Analysis; Ecosystem; Flavanones; Flavonoids; Plant Roots; Plants, Medicinal; Scutellaria baicalensis

Hemin-nitrite-H2O2 system may play a role in liver oxidative injury in some pathological events. In this paper, the effects of the three active components of the root of Scutellaria baicalensis Georgi, i.e. baicalin, baicalein and wogonin, on hemin-nitrite-H2O2 induced liver injury were studied in liver homogenate, liver microsome and human hepatoblastoma cell line HepG2 cells. It was found that hemin-nitrite-H2O2 could induce liver homogenate protein nitration, lipid peroxidation and liver microsome protein oxidation; it also caused a decrease of HepG2 cells viability. Baicalein, baicalin and wogonin could inhibit protein nitration and lipid peroxidation in liver homogenate as well as in HepG2 cells in a dose-dependent manner, the inhibition order was baicalein>baicalin>>wogonin. These three flavonoids also inhibited the oxidation of protein in liver microsome, the decrease of cell viability and the content of GSH in HepG2 cells, among which baicalin represented the most inhibitory effect. Besides, hemin-H2O2 induced cell injury could be augmented with the existence of nitrite, indicating protein nitration involved in hemin-nitrite-H2O2 induced liver injury. These results demonstrated hemin-nitrite-H2O2 could induce liver injury through oxidizing or nitrating different biomolecules. Baicalein, baicalin and wogonin could inhibit hemin-nitrite-H2O2 induced liver injury in dose-dependent manners by inhibiting oxidation and nitration.

Topics: Animals; Cell Line, Tumor; Cell Survival; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Flavanones; Flavonoids; Glutathione; Hemin; Humans; Hydrogen Peroxide; Lipid Peroxidation; Liver; Liver Diseases; Microsomes, Liver; Nitrates; Nitrites; Nitrosation; Oxidation-Reduction; Plant Extracts; Proteins; Rats; Scutellaria baicalensis

Topics: Animals; Cardiotonic Agents; Cells, Cultured; Chickens; Flavanones; Flavonoids; Heart; Seeds

To establish an assay method for the determination of three kinds of biologically active components, five compounds (emodin, chrysophanol, baicalin, wogonin and berberine hydrochloride) simultaneously in Sanhuang tablets.. HPLC was carried out, using a C18 column (150 mm x4. 6 mm ID, 5 microm) set at 30 degrees C, acetonitrile-0.02 mol x L(-1) acetic ammonium (adjusted pH to 3. 50 with acetic acid glacial) as mobile phase (using gradient) with flowing rate 1. 00 mL x min(-1) and detected at 270 nm.. The calibration curve of emodin was linear from 0.020 7 microg to 0.207 microg with r = 0.999 9, the average recovery was 99.65% with RSD 1.25%. The calibration curve of chrysophanol was linear from 0.052 microg to 0.52 microg with r = 0.999 9, and the average recovery was 100.36% with RSD 0.96%. The calibration curve of baicalin was linear from 0.250 5 microg to 2.505 microg with r =0.999 8, and the average recovery was 100.22% with RSD 1.29%. The calibration curve of wogonin was linear from 0.047 6 microg to 0.476 microg with r = 0.999 9, and the average recovery was 98.97% with RSD 1.20%. The calibration curve of berberine hydrochloride was linear from 0.053 12 microg to 0.531 2 microg with r = 0.999 5, and the average recovery was 96.02% with RSD 2.02%. The established method had also been used in the determination of the 5 compounds in 10 different batches of Sanhuang tablets.. This method was proved to be accurate and quick, and can be used for the quality control of the preparation all-around.

Topics: Anthraquinones; Berberine; Chromatography, High Pressure Liquid; Coptis; Drug Combinations; Drugs, Chinese Herbal; Emodin; Flavanones; Flavonoids; Plants, Medicinal; Quality Control; Reproducibility of Results; Rheum; Scutellaria baicalensis; Tablets

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of baicalein, baicalin, oroxylin A and wogonin, Scutellaria baicalensis active components in rat plasma was developed. After liquid-liquid extraction with 2-(3,4-dimethoxy-phenyl)-5,7-dihydroxy-chromen-4-one as internal standard, baicalein, baicalin, oroxylin A and wogonin were eluted from an Atlantis C(18) column within 7 min with isocratic mobile phase consisting of methanol and 0.1% formic acid (60:40, v/v). The analytes were detected using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode. The standard curves were linear (r=1.000) over the concentration ranges of 5-500 ng/ml for baicalein, wogonin and oroxylin A and 5-5000 ng/ml for baicalin. The coefficients of variation and relative errors of baicalein, wogonin, oroxylin A and baicalin for intra- and inter-assay at three or four quality control (QC) levels were 0.8-6.1% and -4.0 to 5.8%, respectively. The lower limits of quantification for baicalein, wogonin, oroxylin A and baicalin were 5ng/ml using 50 microl of plasma sample. This method was successfully applied to the pharmacokinetic study of baicalein, baicalin, wogonin and oroxylin A after an intravenous administration of Scutellariae radix extract to male Sprague-Dawley rats.

Topics: Animals; Chromatography, Liquid; Flavanones; Flavonoids; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Scutellaria; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Microemulsion electrokinetic chromatography (MEEKC) using 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM-BF4) ionic liquid (IL) as additive was developed for the analysis of baicalin, wogonin and baicalein in Scutellariae radix and its preparation. After conducting a series of optimizations, baseline separation was obtained for the analytes within 5min under the optimum conditions (sodium dodecyl sulfate (SDS) 0.88% (m/v) ethyl acetate 0.8% (v/v) butan-1-ol 0.2% (v/v) and the buffer composition were 25% acetonitrile (v/v), 7.5 mM BMIM-BF4 and 10 mM NaH2PO4, pH 8.2, applied voltage 17.5 kV and detection at 254 nm), the method has been successfully applied to the determination and quantification of the analytes in the extracts of S. radix (cooked), S. radix (raw) and Qingfeiyihuowan which was the preparation including S. radix.

Topics: Borates; Chromatography, Micellar Electrokinetic Capillary; Drugs, Chinese Herbal; Flavanones; Flavonoids; Imidazoles; Molecular Structure; Plant Extracts; Reproducibility of Results; Scutellaria baicalensis

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

In vitro manipulation of plant regeneration in the Chinese medicinal species Scutellaria baicalensis Georgi (Huang-qin) resulted in 26 chemically distinct germplasm lines. Antioxidant potential, growth rate and concentration of baicalin, baicalein, melatonin, and wogonin were the selective markers used to identify elite lines. Metabolomic analysis of a subset of the most distinct lines revealed that Huang-qin extracts contained over 2,000 compounds including 781 determined to be of putative medicinal importance as determined by a database search, as well as previously unidentified amino-derivatives of baicalin and wogonin. Huang-qin also contained a metabolite with the same net formula as hyperforin, previously thought to be unique to Hypericum perforatum L. Together these results provide new insights into the biochemical complexity of an important medicinal species and demonstrate the power of in vitro manipulation in combination with untargeted metabolomic screening for the production of new germplasm.

Topics: Antioxidants; Drugs, Chinese Herbal; Flavanones; Flavonoids; Genetic Variation; Melatonin; Scutellaria baicalensis

The objective of the study was to investigate the effects of baicalin, baicalein, and wogonin (plant flavonoids) on interleukin-6 (IL-6) and interleukin-8 (IL-8) protein production, mRNA expression, and nuclear factor-kappaB (NF-kappaB) binding activities induced by interleukin-1beta (IL-1beta) in human retinal pigment epithelial cell line (ARPE-19) cells. To induce IL-6 and IL-8 mRNA expression and protein levels, IL-1beta was added to serum-free medium of ARPE-19 cells and incubated. The flavonoids were added to the medium. IL-6 and IL-8 in the media were measured using enzyme-linked immunosorbent assay. Both IL-6 and IL-8 mRNA were measured by semiquantitative reverse transcription polymerase chain reaction. The binding activities of the transcription factor NF-kappaB complexes to IL-6 and IL-8 were measured by electrophoretic mobility shift assay. IL-6 and IL-8 in the culture media of ARPE-19 cells were increased by IL-1beta in a dose-dependent manner. Baicalin did not suppress IL-1beta-induced IL-6 and IL-8 production, but dexamethasone, baicalein, and wogonin, significantly suppressed IL-6 and IL-8 production. Elevation of IL-6 and IL-8 mRNA was not suppressed by baicalin but was significantly suppressed by dexamethasone, baicalein, and wogonin. NF-kappaB binding activities were not suppressed by baicalin and baicalein, but was suppressed by wogonin. Wogonin and baicalein inhibited IL-1beta-induced IL-6 and IL-8 mRNA and protein production in ARPE-19 cells. The data suggest that wogonin may inhibit IL-6 and IL-8 mRNA expression via the suppression of NF-kappaB binding activities.

Topics: Anti-Inflammatory Agents; Cell Line; Cell Survival; Dexamethasone; Dose-Response Relationship, Drug; Flavanones; Flavonoids; Humans; Interleukin-1; Interleukin-6; Interleukin-8; NF-kappa B; Pigment Epithelium of Eye; Polymerase Chain Reaction; RNA, Messenger

Neuronal injury in the central nervous system following ischemic insult is believed to result from glutamate toxicity and glucose deprivation. In this study, polyphenols isolated from Scutellaria baicalensis Georgi, including baicalin, baicalein, and wogonin, were investigated for their neuroprotective effects against glutamate/NMDA (Glu/NMDA) stimulation and glucose deprivation in primary cultured rat brain neurons. Cell death was accessed by lactate dehydrogenase (LDH) release assay for necrosis, and mitochondrial activity was accessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction activity assay. It was found that both baicalin and baicalein decreased LDH release of the cultured neurons after 24 h treatment, whereas wogonin profoundly increased LDH release after 2 h treatment and resulted in neuronal death after 24 h. Glu/NMDA treatment profoundly increased LDH release and moderately decreased MTT reduction activity in an NMDA receptor-dependent manner. Both baicalin and baicalein significantly reduced Glu/NMDA-increased LDH release, in which baicalein is much more potent than baicalin. Glu/NMDA-increased intracellular calcium was also significantly attenuated by baicalin and baicalein. Baicalin and baicalein did not affect glutamate receptor binding activity, but baicalein did moderately decrease Glu/NMDA-induced nitric oxide (NO) production. In the glucose deprivation (GD) study, baicalein but not baicalin showed significant protective effects on the GD-increased LDH release, without affecting the GD-induced NO production, in cultured rat brain neurons. These results suggest that baicalein is the most effective compound among three polyphenols tested in preventing neurotoxicity induced by both glutamate and GD, whereas baicalin was only effective in preventing glutamate toxicity. Wogonin might have a neurotoxic effect on the brain.

Topics: Animals; Brain Ischemia; Calcium Signaling; Cell Survival; Cells, Cultured; Central Nervous System; Drugs, Chinese Herbal; Excitatory Amino Acid Agonists; Female; Flavanones; Flavonoids; Glucose; Glutamic Acid; L-Lactate Dehydrogenase; Mitochondria; Nerve Degeneration; Neurons; Neuroprotective Agents; Nitric Oxide; Phenols; Polyphenols; Pregnancy; Rats; Rats, Sprague-Dawley; Receptors, Glutamate; Scutellaria baicalensis

Micellar electrokinetic chromatography (MEKC) conditions were developed to analyze the constituents of Scutellariae Radix (SR) and Scutellaria baicalensis roots. Using the MEKC method, the major flavonoid constituents of baicalin, baicalein and wogonin of wild and cultivated S. baicalensis roots were compared. In a preliminary comparison of electropherogram, one special peak was found in a wild sample but not in a 2-year-cultivated one. The compound corresponding to the peak was isolated and identified as a phenylethanoid glycoside, acteoside, by comparing the 1H- and 13C-NMR spectral data with that of the authentic compound. This is the first time acteoside has been isolated from the Scutellaria genus. It could only be found in SR derived from wild S. baicalensis roots and 4-year-cultivated plants, but not in plant materials cultivated for 3 years. Applying the MEKC method established in this study, rapid and simultaneous determinations of acteoside together with 3 flavonoids in samples were achieved. The method can thus be used for the quality control of SR in a shorter analysis period than HPLC.

Topics: Chromatography, High Pressure Liquid; Chromatography, Micellar Electrokinetic Capillary; Flavanones; Flavonoids; Glucosides; Phenols; Plant Roots; Scutellaria; Spectrophotometry, Ultraviolet

To describe the difference between native and nonative herbs by determining contents of seven kinds of flavone for twenty-five samples from seventeen areas.. HPLC. Fluid phase: MEOH-H2O-CH3COOH(ICE) (41:59:0.2) and (50:50:0.2). Detection wavelength: 275.. The contents of baicalin are 6%-9%, wogenin are 2%-8%, baicalein are 0.1%-1.6%, neobaicalein are 0.01%-0.2%, wogonin are 0.01%-0.3%, visidulin and oroxylin are trace amounts or undetected.. The native and nonative herbs have no distinct differce in absolute component ratio. The ratio of baicalin and wogenin is under three. The ratio of baicalin and baicalein, baicalin and wogonin is between twenty and fifty.

Topics: Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Plant Roots; Plants, Medicinal; Quality Control; Scutellaria baicalensis

The effects of the flavonoids from Scutellaria baicalensis Georgi (baicalein, baicalin and wogonin) in cultured human hepatoma cells (Hep G2, Hep 3B and SK-Hep1) were compared by MTT assay and flow cytometry. All three flavonoids dose-dependently decreased the cell viabilities accompanying the collapse of mitochondrial membrane potential and the depletion of glutathione content. However, the influence of baicalein, baicalin or wogonin on cell cycle progression was different. All three flavonoids resulted in prominent increase of G2/M population in Hep G2 cells, whereas an accumulation of sub G1 (hypoploid) peak in Hep 3B cells was observed. In SK-Hep1 cells, baicalein and baicalin resulted in dramatic boost in hypoploid peak, but wogonin made mainly in G1 phase accumulation. These data, together with the previous findings in other hepatoma cell lines, suggest that baicalein, baicalin and wogonin might be effective candidates for inducing apoptosis or inhibiting proliferation in various human hepatoma cell lines.

Topics: Apoptosis; Carcinoma, Hepatocellular; Cell Cycle; DNA, Neoplasm; Drugs, Chinese Herbal; Flavanones; Flavonoids; Flow Cytometry; Glutathione; Humans; Lamiaceae; Medicine, Chinese Traditional; Mitochondria; Molecular Structure; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured

The anti-inflammatory effect of three flavonoids from the root of Scutellaria baicalensis (baicalein, baicalin and wogonin) was evaluated in a murine model of acute experimental colitis induced by dextran sulfate sodium (DSS). Baicalein, but not baicalin or wogonin, given orally at 20 mg/kg for ten days, ameliorates all the considered inflammatory symptoms of the induced colitis, such as body weight loss, blood haemoglobin content, rectal bleeding and other histological and biochemical parameters. The effect of baicalein was similar to that of sulfasalazine, the reference drug given at 50 mg/kg.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Body Weight; Colitis; Dextran Sulfate; Female; Flavanones; Flavonoids; Inflammation; Lamiaceae; Mice; Mice, Inbred BALB C; Plant Extracts; Plant Roots; Sulfasalazine

We previously reported that oroxylin A, a polyphenolic compound, was a potent inhibitor of lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). In the present study, three oroxylin A structurally related polyphenols isolated from the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, were examined for their effects on LPS-induced nitric oxide (NO) production and iNOS and COX-2 gene expressions in RAW 264.7 macrophages. The results indicated that these three polyphenolic compounds inhibited LPS-induced NO production in a concentration-dependent manner without a notable cytotoxic effect on these cells. The decrease in NO production was in parallel with the inhibition by these polyphenolic compounds of LPS-induced iNOS gene expression. However, these three compounds did not directly affect iNOS enzyme activity. In addition, wogonin, but not baicalin or baicalein, inhibited LPS-induced prostaglandin E2 (PGE2) production and COX-2 gene expression without affecting COX-2 enzyme activity. Furthermore, N-nitro-L-arginine (NLA) and N-nitro-L-arginine methyl ester (L-NAME) pretreatment enhanced LPS-induced iNOS (but not COX-2) protein expression, which was inhibited by these three polyphenolic compounds. Wogonin, but not baicalin or baicalein, similarly inhibited PGE2 production and COX-2 protein expression in NLA/LPS or L-NAME/LPS-co-treated RAW 264.7 cells. These results indicated that co-treatment with NOS inhibitors and polyphenolic compounds such as wogonin effectively blocks acute production of NO and, at the same time, inhibits expression of iNOS and COX-2 genes.

Topics: Animals; Antioxidants; Blotting, Western; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Drug Interactions; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Gene Expression; Isoenzymes; Lipopolysaccharides; Macrophages; Mice; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitroarginine; Prostaglandin-Endoperoxide Synthases

To investigate the antitumor effects of Scutellariae radix and its components baicalein, baicalin, and wogonin on human bladder cancer cell lines (KU-1 and EJ-1) and a murine bladder cancer cell line (MBT-2).. Bladder cancer cells were incubated with various concentrations of the agents. Antiproliferative activity against the bladder cancer cell lines was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diplenyl tetrazolium bromide assay. In an in vivo experiment, the mice were subcutaneously injected with MBT-2 cells, and Scutellariae radix was orally administered at a dose of 2 or 10 mg per mouse one time daily for 10 days from day 11 to day 20.. All the drugs inhibited cell proliferation in a dose-dependent manner, but baicalin exhibited the greatest antiproliferative activity. The concentration of baicalin necessary to obtain 50% inhibition was 3.4 microg/mL for KU-1, 4.4 microg/mL for EJ-1, and 0.93 microg/mL for MBT-2. For KU-1 and MBT-2, the percentage of cell survival significantly decreased (P <0.05) at a baicalin concentration of 1 microg/mL. In an in vivo experiment, antitumor effects of Scutellariae radix on C3H/HeN mice implanted with MBT-2 were investigated. All the control mice showed a progressive increase in tumor volume, reaching 2.81 +/- 0.18 cm(3) on day 20 and 5.36 +/- 0.44 cm(3) on day 25. However, when Scutellariae radix was orally administered at a dose of 10 mg per mouse one time daily for 10 days from day 11 to day 20, the tumor volume was 1.99 +/- 0.19 cm(3) on day 20 and 3.86 +/- 0.26 cm(3) on day 25, a significant inhibition of tumor growth (P <0.05). Conclusions. These results suggest that Chinese herbal medicines may become an attractive and promising treatment for bladder cancer.

Topics: Animals; Antineoplastic Agents; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Humans; Male; Mice; Mice, Inbred C3H; Tumor Cells, Cultured

Scutellaria baicalensis (SB) has antibacterial and antiviral activities. Nitric oxide (NO) as a potent macrophage-derived effector molecule against a variety of bacteria, viruses and tumors has received increasing attention. The objective of this study was to determine the effect of SB on the production of NO. Stimulation of mouse peritoneal macrophages with SB after the treatment of recombinant interferon-gamma (rIFN-gamma) resulted in the increased NO production. SB had no effect on NO production by itself. When SB was used in combination with rIFN-gamma, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SB on NO production was shown 6 h after treatment with rIFN-gamma. NO production by SB was inhibited by N(G)-monomethyl-L-arginine. The increased production of NO from rIFN-(gamma) plus SB-stimulated cells was decreased by the treatment of protein kinase C inhibitor such as staurosporin. In addition, synergy between rIFN-gamma and SB was mainly dependent on SB-induced tumor necrosis factor-alpha (TNF-alpha) secretion. All the preparations of SB were endotoxin free. These results suggest that the capacity of SB to increase NO production from rIFN-gamma-primed mouse peritoneal macrophages is the result of SB-induced TNF-gamma secretion.

Topics: Animals; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Interferon-gamma; Macrophages, Peritoneal; Male; Mice; Mice, Inbred C57BL; Nitric Oxide; omega-N-Methylarginine; Recombinant Proteins; Tumor Necrosis Factor-alpha

An ion-pair high-performance liquid chromatographic method for the simultaneous determination of four flavonoids, namely baicalin, wogonoside, baicalein and wogonin, and four berberine-type alkaloids, namely berberine, coptisine, palmatine and jateorrhizine, and glycyrrhizin in Kampo medicines is described. The analysis can be accomplished within 30 min with a Wakosil-II 5C18 HG column by linear gradient elution using a mobile phase containing aqueous phosphoric acid, sodium dodecyl sulfate and acetonitrile at a flow-rate of 1.0 ml x min(-1), a thermostatic oven at 45 degrees C, and detection at 265 nm. The method was applied to quantifying these components in three Kampo decoctions: Oren-gedoku-to, San'o-shashin-to and Hange-shashin-to. The decoctions were diluted with 65% methanol at the final stage because a large quantity of precipitate, mainly from baicalin and berberine, was formed. The within-day relative standard deviations were less than 2.02% (n=10). The recoveries of these compounds were 90.3-102%. The detection limits of these compounds were 0.02-1.96 microM per injection (5 microl).

Topics: Alkaloids; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Flavanones; Flavonoids; Glycyrrhizic Acid; Medicine, Kampo

Free radical scavenging and antioxidant activities of baicalein, baicalin, wogonin and wogonoside, the four major flavonoids in the radix of Scutellaria baicalensis Georgi, were examined in different systems. ESR results showed that baicalein and baicalin scavenged hydroxyl radical, DPPH radical and alkyl radical in a dose-dependent manner, while wogonin and wogonoside showed subtle or no effect on these radicals. Ten micromol/l of baicalein and baicalin effectively inhibited lipid peroxidation of rat brain cortex mitochondria induced by Fe(2+)-ascorbic acid, AAPH or NADPH, while wogonin and wogonoside showed significant effects only on NADPH-induced lipid peroxidation. In a study on cultured human neuroblastoma SH-SY5Y cells system, it was found that 10 micromol/l of baicalein and baicalin significantly protected cells against H(2)O(2)-induced injury. Baicalein was the most effective antioxidant among the four tested compounds in every system due to its o-tri-hydroxyl structure in the A ring. Compared with a well-known flavonoid, quercetin, the antioxidant activity of baicalein was lower in DPPH or AAPH system, but a little higher in those systems which might associate with iron ion. These results suggest that flavonoids in the radix of Scutellaria baicalensis with o-di-hydroxyl group in A the ring, such as baicalein and baicalin, could be good free radical scavengers and might be used to cure head injury associated with free radical assault.

Topics: Animals; Antioxidants; Binding Sites; Brain; China; Drugs, Chinese Herbal; Electron Spin Resonance Spectroscopy; Flavanones; Flavonoids; Free Radical Scavengers; Humans; Hydrogen Peroxide; Hydroxyl Radical; Lipid Peroxidation; Mitochondria; NADP; Plants, Medicinal; Rats; Tumor Cells, Cultured

Inhibitory mechanism of the water extract of Scutellariae Radix on prostaglandin E2 (PGE2) release was examined in C6 rat glioma cells. Scutellariae Radix reduced a Ca2+ ionophore A23187-induced PGE2 release by inhibition of arachidonic acid (AA) liberation. Sho-saiko-to and San'o-shashin-to, which contain Scutellariae Radix, also inhibited PGE2 release. A23187 caused phosphorylation of mitrogen-activated protein kinase (MAPK), resulting in activation of cytosolic phospholipase A2 (cPLA2). Scutellariae Radix and baicalein inhibited the phosphorylation of MAPK. Baicalein, but not baicalin, inhibited A23187-induced PGE2 release. These results suggest that baicalein in Scutellariae Radix reduces AA liberation through the inhibition of the MAPK-cPLA2 pathway.

Topics: Amino Acid Sequence; Animals; Antineoplastic Agents; Arachidonic Acid; Calcimycin; Calcium-Calmodulin-Dependent Protein Kinases; Dinoprostone; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavanones; Flavonoids; Glioma; Ionophores; Molecular Sequence Data; Phosphorylation; Prostaglandin Antagonists; Rats; Tumor Cells, Cultured

The effect of extracts of scutellariae radix (Scutellaria baicalensis Georgi) and its flavonoids, baicalin, baicalein and wogonin, on induction of quinone reductase (QR) in the Hepa 1c1c7 murine hepatoma cell line was examined. A significant and dose-dependent induction of QR activity was observed in the methanol extract of scutellariae radix and baicalin. HPCL analysis showed that baicalin was contained as a main component in the methanol extract of scutellariae radix, indicating that baicalin may be the major active principle of QR induction mediated by scutellariae radix extract. To elucidate the mechanism of baicalin-mediated induction of QR enzyme activity, the effect on QR mRNA levels in Hepa 1c1c7 cell cultures was investigated. Using reverse transcriptase-polymerase chain reaction techniques, time- and dose-dependent induction of QR mRNA levels by baicalin were demonstrated in Hepa 1c1c7 cells. On the basis of these results, the scutellariae radix extract or baicalin can be regarded as a readily available, promising, novel cancer chemopreventive agent.

Topics: Animals; Anticarcinogenic Agents; Base Sequence; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; Enzyme Induction; Flavanones; Flavonoids; Mice; Molecular Sequence Data; NAD(P)H Dehydrogenase (Quinone); Tumor Cells, Cultured

High-performance liquid chromatography was employed to determine the contents of several marker substances such as gentiopicroside, mangiferin, palmatine, berberine, baicalin, wogonin and glycyrrhizin in Sann-Joong-Kuey-Jian-Tang. The separation was performed on a Cosmosil 5C18-AR column by gradient elution with 0.03% (v/v) phosphoric acid-acetonitrile (0 min, 90:10; 10 min, 87:13; 17-27 min, 77:23; 40 min, 62:38; 50 min, 55:45) as the mobile phase at a flow-rate of 1.0 ml/min, with detection at 254 nm. n-Propylparaben was used as the internal standard and seven regression equations revealed linear relationships between the peak-area ratios (marker substances/internal standard) and concentrations. The repeatability and reproducibility (relative standard deviation) of the method were in the ranges 0.02-1.78% and 1.44-4.95%, respectively.

Topics: Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Iridoid Glucosides; Iridoids; Pyrans; Reproducibility of Results; Xanthenes; Xanthones

Five extracts (n-hexane, chloroform, ethyl acetate, n-butanol and water) of Scutellaria rivularis Benth. were evaluated for their anti-inflammatory activity against carrageenan-induced paw edema in rats and compared with indomethacin. The result indicated that chloroform extract proved to be the most effective in all of the extracts. Consequently, three major components (baicalin, baicalein and wogonin) of the chloroform extract were further tested for their anti-inflammatory activity using the same model. It was found that baicalin exhibits the greatest inhibition activity against carrageenan-induced rat paw edema.

Topics: Animals; Anti-Infective Agents; Carrageenan; Flavanones; Flavonoids; Inflammation; Male; Medicine, Chinese Traditional; Pain Measurement; Rats; Rats, Wistar

The effects of baicalein, baicalin and wogonin, the flavonoids from Scutellaria baicalensis, on the proliferative responses of cultured rabbit vascular smooth muscle cells were studied. The proliferative response was determined from the uptake of tritiated thymidine. In rabbit vascular smooth muscle cells, all three flavonoids dose dependently inhibited the proliferative response induced by 5% fetal calf serum at the dose range of 10(-6) to 10(-4) M. Baicalin and wogonin were less effective than baicalein as inhibitors of the serum-induced smooth muscle cell proliferation, indicating that the three hydroxyl groups on positions 5, 6 and 7 seem to be necessary and sufficient for full inhibitory activity against the proliferative response of smooth muscle cells. Baicalein had a greater inhibitory effect on the proliferative reponse stimulated by platelet-derived growth factor than on serum-stimulated proliferation. Baicalein, a flavonoid with antiproliferative and lipoxygenase-inhibitory activities, may be useful as another template for the development of better drugs to prevent the pathological changes of atherosclerosis and restenosis.

Topics: Animals; Cell Division; Cells, Cultured; Depression, Chemical; Drugs, Chinese Herbal; Flavanones; Flavonoids; Muscle, Smooth, Vascular; Platelet-Derived Growth Factor; Rabbits; Thymidine