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Find video protocols related to scientific articles indexed in Pubmed.
Toxicity assessment of Arisaematis Rhizoma in rats by a (1)H NMR-based metabolomics approach.
Mol Biosyst
PUBLISHED: 11-20-2014
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Arisaematis Rhizoma (AR), a famous traditional Chinese medicine, has been widely used in Asia over thousands of years. Documented with noticeable toxicity in ancient books, AR has been used to treat various diseases in the clinic. Therefore, it is important to assess the toxicity of AR dynamically and holistically. In this study, a (1)H NMR-based metabolomics approach complemented with serum chemistry and histopathology has been applied to investigate the toxicity of AR. Rats were intragastrically administered with AR (0, 0.5 and 1 g kg(-1) body weight) for 30 days, and serum and urine samples were collected. Their (1)H NMR profiles were analyzed by multivariate pattern recognition techniques to denote metabolic variations induced by AR, and 13 metabolites in urine and 6 metabolites in serum were significantly altered, which suggested that disturbances in energy metabolism, perturbation of the gut microflora environment, membrane damage, folate deficiency and injury of kidneys are produced by AR. Histopathology showed a slight vacuolization of the glomerular matrix and edema of renal tubular epithelial cells in kidneys of AR administered rats, which were evidenced by increased levels of blood urea nitrogen and creatinine in serum chemistry. Our results indicated that oral administration of crude AR was found to induce slight renal toxicity. Therefore, precautions should be made to monitor the potential nephrotoxicity of AR in clinical use. The metabolomics approach provided a promising tool for the study and better understanding of TCM-induced toxicity dynamically and holistically.
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Simultaneous enrichment and separation of flavonoids from Herba Epimedii by macroporous resins coupled with preparative chromatographic method.
Nat. Prod. Res.
PUBLISHED: 10-03-2014
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An efficient, feasible enrichment and separation method of epimedins A, B, C and icariin from Herba Epimedii was developed by the combination of microwave-assisted extraction, macroporous resins and preparative HPLC. WDX-5 macroporous resin shows better recoveries at 96.2%, 97.0%, 98.2% and 97.1% for epimedins A, B, C and icariin than other macroporous resins used in the experiments. As a result, epimedins A (5.1 mg), B (15.3 mg), C (7.6 mg) and icariin (14.3 mg) were obtained from 6.0 g crude Herba Epimedii with the recoveries at 70.8%, 68.9%, 66.7% and 95.3%, respectively. The method developed in this study may provide scientific references for the enrichment and separation of flavonoids from Herba Epimedii.
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The effects of nodakenin on airway inflammation, hyper-responsiveness and remodeling in a murine model of allergic asthma.
Immunopharmacol Immunotoxicol
PUBLISHED: 08-04-2014
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Nodakenin is a major coumarin glucoside in the root of Peucedanum decursivum Maxim, a commonly used traditional Chinese medicine for the treatment of asthma and chronic bronchitis for thousands of years.
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Analysis and pharmacokinetics studies of gastrodin and p-hydroxybenzyl alcohol in dogs using ultra fast liquid chromatography-tandem mass spectrometry method.
J Pharm Biomed Anal
PUBLISHED: 07-12-2014
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A simple, sensitive and reliable ultra fast liquid chromatography-electrospray ionization-tandem mass spectrometry (UFLC-ESI-MS/MS) method was developed for simultaneously quantifying gastrodin (p-hydroxy-methyl-phenol-?-d-glucoside) and its metabolite p-hydroxybenzyl alcohol (HBA) in dog plasma. Separation was performed on an ultra fast liquid chromatography (UFLC) system. Detection was carried out on a tandem mass spectrometry (MS/MS) in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) interface. MRM mode of precursor-product ion transitions was used for gastrodin, HBA and the internal standard (IS, bergeninum) at m/z 285.0?123.0, 123.0?105.0 and 326.9?192.2, respectively. The lower limits of quantification (LLOQ) of this method for both gastrodin and HBA were 1ng/mL, with their linear concentration ranging from 0.001 to 10?g/mL. The methods were validated for selectivity, calibration curves, accuracy and precision, extraction recoveries, matrix effects, carry-over, cross talk, dilution integrity, stability and incurred sample reanalysis (ISR). Using this validated method, pharmacokinetic behaviors of gastrodin and HBA after intragastric administration (ig) of gastrodin to dogs were studied for the first time.
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A new acorane sesquiterpene from the aerial parts of Psychotria yunnanensis.
Nat. Prod. Res.
PUBLISHED: 07-10-2014
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Psycacoraone A (1), a new acorane-type sesquiterpene possessing rare spirobicyclic carbon skeleton, was isolated from the aerial parts of Psychotria yunnanensis. Its structure was elucidated on the basis of spectroscopic methods including HR-ESI-MS, 1D and 2D NMR. The absolute configuration of 1 was determined by calculation of electronic circular dichroism using density functional theory.
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NMR-based metabolomics approach to study the chronic toxicity of crude ricin from castor bean kernels on rats.
Mol Biosyst
PUBLISHED: 07-04-2014
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Ricin, a large, water soluble toxic glycoprotein, is distributed majorly in the kernels of castor beans (the seeds of Ricinus communis L.) and has been used in traditional Chinese medicine (TCM) or other folk remedies throughout the world. The toxicity of crude ricin (CR) from castor bean kernels was investigated for the first time using an NMR-based metabolomic approach complemented with histopathological inspection and clinical chemistry. The chronic administration of CR could cause kidney and lung impairment, spleen and thymus dysfunction and diminished nutrient intake in rats. An orthogonal signal correction partial least-squares discriminant analysis (OSC-PLSDA) of metabolomic profiles of rat biofluids highlighted a number of metabolic disturbances induced by CR. Long-term CR treatment produced perturbations on energy metabolism, nitrogen metabolism, amino acid metabolism and kynurenine pathway, and evoked oxidative stress. These findings could explain well the CR induced nephrotoxicity and pulmonary toxicity, and provided several potential biomarkers for diagnostics of these toxicities. Such a (1)H NMR based metabolomics approach showed its ability to give a systematic and holistic view of the response of an organism to drugs and is suitable for dynamic studies on the toxicological effects of TCM.
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(1)H NMR-based metabolomics study on a goldfish model of Parkinson's disease induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP).
Chem. Biol. Interact.
PUBLISHED: 03-26-2014
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A goldfish (Carassius auratus) model of Parkinson's disease (PD) was constructed by a single dose of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) according to previously reported methods. Global metabolite changes in brain of the MPTP induced goldfish model of PD were investigated. (1)H NMR-based metabolomics combined with various statistical methods such as orthogonal partial least squares discriminant analysis (OPLS-DA) and two-dimensional statistical total correlation spectroscopy (2D-STOCSY) found significant increase of leucine, isoleucine, valine, alanine, alanylalanine, creatinine, myo-inositol, 18:2 fatty acid, total fatty acids, arachic alcohol, taurine and significant decrease of N-acetylaspartate, (phospho)creatine, (phospho)choline, betaine, glutamine, 3-hexenedioate, acetamide, malonate, isocitrate, scyllo-inositol, phosphatidylcholines, cholesterols, n-3 fatty acids, polyunsaturated fatty acids (PUFAs) in brain of MPTP induced PD goldfish. These disturbed metabolite levels were involved in oxidative stress, energy failure, neuronal cell injury and death, consistent with those observed in clinical PD patients, and rodents and primates model of PD, indicating that the acute MPTP model of goldfish was an ideal and valuable model for PD research. In addition, several unusual metabolites in brain were significantly changed between MPTP induced PD and control goldfish, which might also play an important role in the pathogenesis of PD. This study also demonstrated the applicability and potential of (1)H NMR-based metabolomics approach for evaluation of animal models of disease induced by chemicals, such as MPTP-induced PD goldfish.
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Baicalin inhibits hypoxia-induced pulmonary artery smooth muscle cell proliferation via the AKT/HIF-1?/p27-associated pathway.
Int J Mol Sci
PUBLISHED: 03-25-2014
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Baicalin, a flavonoid compound purified from the dry roots of Scutellaria baicalensis Georgi, has been shown to possess various pharmacological actions. Previous studies have revealed that baicalin inhibits the growth of cancer cells through the induction of apoptosis. Pulmonary arterial hypertension (PAH) is a devastating disease characterized by enhanced pulmonary artery smooth muscle cell (PASMCs) proliferation and suppressed apoptosis. However, the potential mechanism of baicalin in the regulation of PASMC proliferation and the prevention of cardiovascular diseases remains unexplored. To test the effects of baicalin on hypoxia, we used rats treated with or without baicalin (100 mg·kg?¹ each rat) at the beginning of the third week after hypoxia. Hemodynamic and pulmonary pathomorphology data showed that right ventricular systolic pressures (RVSP), the weight of the right ventricle/left ventricle plus septum (RV/LV + S) ratio and the medial width of pulmonary arterioles were much higher in chronic hypoxia. However, baicalin treatment repressed the elevation of RVSP, RV/LV + S and attenuated the pulmonary vascular structure remodeling (PVSR) of pulmonary arterioles induced by chronic hypoxia. Additionally, baicalin (10 and 20 ?mol·L?¹) treatment suppressed the proliferation of PASMCs and attenuated the expression of hypoxia-inducible factor-? (HIF-?) under hypoxia exposure. Meanwhile, baicalin reversed the hypoxia-induced reduction of p27 and increased AKT/protein kinase B phosphorylation p-AKT both in vivo and in vitro. These results suggested that baicalin could effectively attenuate PVSR and hypoxic pulmonary hypertension.
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Identification and analysis of gastrodin and its five metabolites using ultra fast liquid chromatography electrospray ionization tandem mass spectrometry to investigate influence of multiple-dose and food.
J Chromatogr A
PUBLISHED: 03-04-2014
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A reliable and highly sensitive ultra performance liquid chromatography electrospray ionization tandem mass spectrometry (UFLC-ESI-MS/MS) analytical method was developed for identification and quantification of gastrodin (GAS) and its metabolites in rat plasma. Five metabolites were identified: p-formylphenyl-?-d-glucopyranoside (M1), p-hydroxybenzonic acid (M2), p-hydroxybenzyl alcohol (M3), p-formaldehydephenyl-?-d-glucopyranoside (M4), p-hydroxybenzaldehyde (M5). The molecular structures of metabolites were proposed based on the characters of their precursor ions, product ions and chromatographic retention time. Four of them were reported firstly in rat plasma. This method involved the addition of bergeninum as the internal standard (IS), UFLC separation, and quantification by MS/MS system using negative electrospray ionization in the multiple reaction monitoring (MRM) mode. The lower limit of quantification of gastrodin and five metabolites were all 1ng/mL. The method was linear in the concentration range of 0.001-10?g/mL. The intra- and inter-day precisions (R.S.D %) were within 15.0% for all analytes. No interference was noted due to endogenous substances. All analytes were stable in rat plasma stored at room temperature and 4°C for at least 4h, -20°C combined with three freeze-thaw cycles for at least 1 month. By this method, the influence of multiple-dose and food on the pharmacokinetics behaviors of GAS and its metabolites were studied for the first time. We hope pharmacokinetic data of present study may inspire rational clinical usage of GAS.
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NMR-based metabolomics approach to study the toxicity of lambda-cyhalothrin to goldfish (Carassius auratus).
Aquat. Toxicol.
PUBLISHED: 08-30-2013
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In this study, a (1)H nuclear magnetic resonance (NMR) based metabolomics approach was applied to investigate the toxicity of lambda-cyhalothrin (LCT) in goldfish (Carassius auratus). LCT showed tissue-specific damage to gill, heart, liver and kidney tissues of goldfish. NMR profiling combined with statistical methods such as orthogonal partial least squares discriminant analysis (OPLS-DA) and two-dimensional statistical total correlation spectroscopy (2D-STOCSY) was developed to discern metabolite changes occurring after one week LCT exposure in brain, heart and kidney tissues of goldfish. LCT exposure influenced levels of many metabolites (e.g., leucine, isoleucine and valine in brain and kidney; lactate in brain, heart and kidney; alanine in brain and kidney; choline in brain, heart and kidney; taurine in brain, heart and kidney; N-acetylaspartate in brain; myo-inositol in brain; phosphocreatine in brain and heart; 2-oxoglutarate in brain; cis-aconitate in brain, and etc.), and broke the balance of neurotransmitters and osmoregulators, evoked oxidative stress, disturbed metabolisms of energy and amino acids. The implication of glutamate-glutamine-gamma-aminobutyric axis in LCT induced toxicity was demonstrated for the first time. Our findings demonstrated the applicability and potential of metabolomics approach for the elucidation of toxicological effects of pesticides and the underlying mechanisms, and the discovery of biomarkers for pesticide pollution in aquatic environment.
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Labdane diterpenes from Chloranthus serratus.
Fitoterapia
PUBLISHED: 06-27-2013
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Five new labdane diterpenes (1-5), serralabdanes A-E, were isolated from the whole plant of Chloranthus serratus. Their structures were elucidated by spectroscopic methods, and the absolute configuration of the 12,13-diol moiety in serralabdane C (3) was determined by observing the induced circular dichroism (ICD) after addition of dimolybdenum teracetate in DMSO solution. Serralabdanes A-E (1-5) showed inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW264.7 cells.
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Pentasaccharide resin glycosides from Ipomoea cairica and their cytotoxic activities.
Phytochemistry
PUBLISHED: 06-27-2013
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Six partially acylated pentasaccharide resin glycosides, cairicosides A-F, were isolated from the aerial parts of Ipomoea cairica. These compounds were characterized as a group of macrolactones of simonic acid A, partially acylated with different organic acids. The lactonization site of 11S-hydroxyhexadecanoic acid (jalapinolic acid) was bound to the second saccharide moiety at C-3 in cairicosides A-E, while at C-2 in cairicoside F. Structures were established by spectroscopic and chemical methods. Compounds cairicosides A-E exhibited moderate cytotoxicity against a small panel of human tumor cell lines with IC50 values in the range of 4.28-14.31?M.
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¹H NMR-based metabolomics approach to evaluate the effect of Xue-Fu-Zhu-Yu decoction on hyperlipidemia rats induced by high-fat diet.
J Pharm Biomed Anal
PUBLISHED: 02-09-2013
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An NMR-based metabolomics approach was conducted to holisticly explore the effect of XFZYD (a traditional Chinese medicine formula) on high-fat diet induced hyperlipidemia rats with one of the commonly used antihyperlipidemic agents, simvastatin as the positive control. NMR spectra from blood plasma combined with statistical analysis revealed compounds distinguishing hyperlipidemia rats from normal control rats. XFZYD could ameliorate hyperlipidemia by intervening in some major metabolic pathways, such as decreasing the accumulation of ketone body (?-hydroxybutyrate) and acetyl-glycoproteins, enhancing glutathione (GSH) biosynthesis, partially reversing energy and lipid metabolism disturbance. Oral administration of XFZYD could also be helpful to hyperlipidemia rats in bettering the serum chemistry profile. The combined results demonstrated that XFZYD could ameliorate the hyperlipidemic symptoms in a global scale and restore the abnormal metabolic state to a near normal level in a time-dependent pattern.
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A rapid and sensitive LC-MS/MS assay for the determination of berbamine in rat plasma with application to preclinical pharmacokinetic study.
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
PUBLISHED: 02-04-2013
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Berbamine (BBM), a natural compound from Chinese herb Berberis amurensis, has recently received a great deal of attention due to its anti-leukemia activity. In this study, a rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of BBM in rat plasma was developed for the first time. Caffeine was used as an internal standard. Chromatographic separation was performed on an ODS column with gradient elution using methanol-1% formic acid as mobile phase at a flow rate of 0.3mL/min. Quantification was through tandem mass spectrometry with positive electrospray ionization (ESI) and multiple reaction monitoring (MRM) at m/z 305.2?566.3 and 195.1?138.0 for BBM and IS, respectively. The lower limit of quantification was 1ng/mL with a linear range of 1-1000ng/mL. The intra- and inter-day assay precision (RSD) ranged from 2.0-6.4% to 2.5-5.5%, respectively, and the intra- and inter-day assay accuracy (RE) was between -5.8-6.0% and -6.5-1.4%, respectively. The validated method was successfully applied to the preclinical pharmacokinetic studies of BBM in rats. The elimination half-lives (t1/2) were (472.4±66.1), (509.6±97.0) and (486.2±94.6) min after single intravenous administration of 2, 4 and 8mg/kg BBM, respectively. The area under the plasma concentration versus time curve (AUC0-24h) and initial plasma concentration (C0) were linearly related to dose.
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A three-dimensional water quality modeling approach for exploring the eutrophication responses to load reduction scenarios in Lake Yilong (China).
Environ. Pollut.
PUBLISHED: 01-16-2013
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Lake Yilong in Southwestern China has been under serious eutrophication threat during the past decades; however, the lake water remained clear until sudden sharp increase in Chlorophyll a (Chl a) and turbidity in 2009 without apparent change in external loading levels. To investigate the causes as well as examining the underlying mechanism, a three-dimensional hydrodynamic and water quality model was developed, simulating the flow circulation, pollutant fate and transport, and the interactions between nutrients, phytoplankton and macrophytes. The calibrated and validated model was used to conduct three sets of scenarios for understanding the water quality responses to various load reduction intensities and ecological restoration measures. The results showed that (a) even if the nutrient loads is reduced by as much as 77%, the Chl a concentration decreased only by 50%; and (b) aquatic vegetation has strong interaction with phytoplankton, therefore requiring combined watershed and in-lake management for lake restoration.
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Complete 1H and 13C NMR data assignment of protolimonoids from the stem barks of Aphanamixis grandifolia.
Magn Reson Chem
PUBLISHED: 04-20-2011
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Seven new protolimonoids, named aphagranins A-G (1-7), along with four known compounds, were isolated from the ethanol extract of the stem barks of Aphanamixis grandifolia. Structure elucidation and signal assignments were achieved on the basis of spectral and chemical evidences.
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Inhibition of P-glycoprotein at the blood-brain barrier by phytochemicals derived from traditional Chinese medicine.
Cancer Genomics Proteomics
PUBLISHED: 07-27-2010
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The blood-brain barrier (BBB) is a key determinant for drug transport through brain vessels. It restricts the pharmacological efficacy in numerous neurological diseases, including brain tumors. A major functional constituent of BBB is P-glycoprotein, which is also a major obstacle for effective chemotherapy of brain tumors. An appealing strategy is to selectively modulate BBB function using P-glycoprotein inhibitors. We assessed 57 chemically defined compounds derived from medicinal plants used in traditional Chinese medicine for their potential to inhibit P-glycoprotein. Nine phytochemicals inhibited P-glycoprotein in porcine brain capillary endothelial cells (PBCECs) and multidrug-resistant CEM/ADR5000 cells as shown by a calcein fluorescence assay. The cytotoxicity of the 57 phytochemicals was measured by a growth inhibition assay. Seven compounds inhibiting P-glycoprotein at lower doses were cytotoxic to drug-sensitive parental CCRF-CEM cells at higher doses. Of them, five were not cross-resistant to CEM/ADR5000 cells (baicalein, bufalin, glybomine B, deoxyserofendic acid, and shogaol). Bufalin was chosen as a lead compound. Of a further six bufalin-related compounds, scillarenin showed improved features in comparison to bufalin. It was cytotoxic to cancer cells at a nanomolar range. COMPARE and hierarchical cluster analyses of microarray-based mRNA expression were used to investigate determinants of sensitivity or resistance of the bufalin-related compounds downstream of P-glycoprotein. CEM/ADR5000 cells were not cross-resistant, but were collaterally sensitive towards scillarenin. Finally, scillarenin inhibited P-glycoprotein in PBCECs. Taken together, these data show that scillarenin is a potential novel candidate for P-glycoprotein inhibition at BBB, and, thereby, may improve the efficacy of therapy regimens in treating brain diseases.
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Cytotoxic tirucallane C26 triterpenoids from the stem barks of Aphanamixis grandifolia.
Phytochemistry
PUBLISHED: 03-26-2010
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Five tirucallane type C(26) triterpenoids, accompanied by two known compounds, 3?-hydroxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone and 3-oxo-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone, were isolated from the stem barks of Aphanamixis grandifolia. Their structures were established mainly by means of a combination of 1D and 2D NMR spectroscopic and mass spectrometry techniques as 3?-hydroxyl-21?-methoxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone, 3?-hydroxy-21?-methoxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone, 3-oxo-21?-methoxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone, 3-oxo-21?-methoxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone, and 3-oxo-21?-ethoxy-24,25,26,27-tetranortirucall-7-ene-23(21)-lactone. All isolates were in vitro evaluated for their cytotoxic activities against five tumor cell lines (MCF-7, HeLa, HepG2, SGC-7901 and BGC-823).
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Quantitative analysis of four major diterpenoids in Andrographis paniculata by 1H NMR and its application for quality control of commercial preparations.
J Pharm Biomed Anal
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A quantitative proton nuclear magnetic resonance technique (qHNMR) has been successfully introduced to quantify andrographolide, dehydroandrographolide, deoxyandrographolide and neoandrographolide in Andrographis paniculata, a commonly used important traditional Chinese medicine. Creative use of trifluoroacetic acid-d, which satisfactorily resolved the overlapping signals of these compounds in crowded regions of ? 4.5-5.6 ppm in (1)H NMR spectrum, made their quantification possible. Optimization of other experimental conditions, including internal standard, NMR pulse sequence, and NMR relaxation delay time, finally established the (1)H NMR based quantification approach, which was validated with satisfactory accuracy, precision, repeatability, and recovery. Except for deoxyandrographolide and neoandrographolide in two compound recipes, this method was successfully applied to quantify the four major components in fourteen raw herb materials and five commercial preparations, providing quantification results in good agreement with those determined by HPLC. The inherent advantages of qHNMR, such as its rapidity and simplicity, make itself a feasible alternative to HPLC for the quality control of A. paniculata raw material and herbal preparations.
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Effects of (±)-praeruptorin A on airway inflammation, airway hyperresponsiveness and NF-?B signaling pathway in a mouse model of allergic airway disease.
Eur. J. Pharmacol.
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The root of Peucedanum praeruptorum Dunn is a traditional Chinese medicine commonly used to treat asthma in China. (±)-praeruptorin A (PA) is the most abundant constituent of P. praeruptorum Dunn, the effects of which on asthma were investigated using a murine model of allergic airway disease. BALB/c mice were sensitized and challenged by ovalbumin to induce airway inflammation. PA was administered intragastrically before every OVA challenge. Airway responsiveness was measured by a lung function analysis system. The number of total leukocytes in bronchoalveolar lavage fluid was counted using a hemocytometer, and differential cell counts were determined using Diff-Quick-stained smears. Histopathology of lung tissue was analyzed by hematoxylin-eosin and Congo red staining. Levels of inflammatory mediators in bronchoalveolar lavage fluid and immunoglobulins in serum were measured by enzyme-linked immunosorbent assay. The expression of pulmonary eotaxin was detected by immunohistochemistry and reverse transcription polymerase chain reaction. The activation of NF-?B was evaluated by electrophoretic mobility shift assay and western blot analysis. Compared with model group, PA significantly reduced airway hyperresponsiveness and airway eosinophilic inflammation, improved pathologic lesion of the lungs, reduced levels of interleukin (IL)-4, IL-5, IL-13 and LTC? in bronchoalveolar lavage fluid and immunoglobulin (Ig) E in serum, and inhibited eotaxin protein and mRNA expression, I?B? degradation, NF-?B nuclear translocation, NF-?B DNA-binding activity and RelA/p65 phosphorylation in lung, which suggested that PA can significantly suppress OVA-induced airway inflammation and airway hyperresponsiveness in mice, showing great therapeutic potential for the treatment of allergic asthma.
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