ABSTRACT
Chrysanthemum boreale is a plant widespread in East Asia, used in folk medicine to treat various disorders, such as pneumonia, colitis, stomatitis, and carbuncle. Whether the essential oil from C. boreale (ECB) and its active constituents have anti-proliferative activities in lung cancer is unknown. Therefore, we investigated the cytotoxic effects of ECB in A549 and NCI-H358 human lung cancer cells. Culture of A549 and NCI-H358 cells with ECB induced apoptotic cell death, as revealed by an increase in annexin V staining. ECB treatment reduced mitochondrial membrane potential (MMP), disrupted the balance between pro-apoptotic and anti-apoptotic Bcl-2 proteins, and activated caspase-8, -9, and -3, as assessed by western blot analysis. Interestingly, pretreatment with a broad-spectrum caspase inhibitor (z-VAD-fmk) significantly attenuated ECB-induced apoptosis. Furthermore, gas chromatography-mass spectrometry (GC/MS) analysis of ECB identified six compounds. Among them, ß-caryophyllene exhibited a potent anti-proliferative effect, and thus was identified as the major active compound. ß- Caryophyllene induced G1 cell cycle arrest by downregulating cyclin D1, cyclin E, cyclin-dependent protein kinase (CDK) -2, -4, and -6, and RB phosphorylation, and by upregulating p21CIP1/WAF1 and p27KIP1. These results indicate that ß-caryophyllene exerts cytotoxic activity in lung cancer cells through induction of cell cycle arrest.
Subject(s)
Cell Cycle Proteins/metabolism , Chrysanthemum/chemistry , Lung Neoplasms/metabolism , Polycyclic Sesquiterpenes/pharmacology , A549 Cells , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Humans , Lung Neoplasms/drug therapy , Membrane Potential, Mitochondrial/drug effects , Oils, Volatile/pharmacologyABSTRACT
[This corrects the article DOI: 10.1155/2013/859624.].
ABSTRACT
A new C-glycosylflavone, drymaritin E (6-C-(3-keto-ß-digitoxopyranosyl)-4'-O-(ß-D-glucopyranosyl)-7-methoxyl-5,4'-dihydroxylflavone) 1 was isolated from the oily upper phase (SU) of the MeOH extract from aerial parts of Drymaria cordata together with two known compounds (cassiaoccidentalin A 2 and anemonin 3) and an inseparable mixture of two known C-glycosylflavones 5,4'-dihydroxy-7-methoxyflavone-6-C-(2''-O-α-L-rhamnopyranosyl)-ß-D-glucopyranoside 4a and 5,7,3',4'-tetrahydroxyflavone-6-C-(2''-O-α-L-rhamnopyranosyl)-ß-D-glucopyranoside 4b. The alkaline hydrolysis of 3 led to a new hemisynthetic derivative, sodium anemonate (sodium 2-((1'E) 2'-sodium-carboxylate-vinyl)-5-oxo-cyclohex-1-ene carboxylate) 3a. The chemical structures were determined by spectroscopic methods ((1)H NMR, (13)C NMR, (1)H-(1)H COSY, HMBC, HSQC, and NOESY) and mass spectrometry (ESI-MS). C-glycosylflavones had significant free radical-scavenging activities on the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). However, SU and compounds 3 and 3a exhibited no activity. In particular, compound 1 exhibited a concentration-dependent radical scavenging activity on DPPH with EC50 of 31.43 µg/mL.
Subject(s)
Antioxidants/pharmacology , Caryophyllaceae , Flavonoids/pharmacology , Plant Extracts/pharmacology , Animals , Antioxidants/chemistry , Antioxidants/isolation & purification , Dose-Response Relationship, Drug , Erythrocytes/drug effects , Erythrocytes/metabolism , Flavones/chemistry , Flavones/isolation & purification , Flavones/pharmacology , Flavonoids/chemistry , Flavonoids/isolation & purification , Male , Plant Components, Aerial , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Rats , Rats, WistarABSTRACT
Phytochemical studies on the constituents of the rhizomes of Imperata cylindrica (Gramineae) were performed using high-performance liquid chromatography (HPLC). We also aimed to search for any biologically active substance capable of inhibiting nitric oxide (NO) formation in lipopolysaccharide (LPS)-activated macrophage 264.7 cells, by testing four compounds isolated from this plant. Four compounds, including a new chromone, isoeugenin, along with ferulic acid, p-coumaric acid, and caffeic acid were isolated and identified by NMR spectroscopy. The structure of isoeugenin was determined as 7-hydroxy-5-methoxy-2-methylchromone by the 2D-NMR technique. Among the four compounds, isoeugenin has the lowest IC50 value on the inhibition of NO production in LPS-activated macrophage RAW264.7 cells (IC50, 9.33 µg/mL). In addition, isoeugenin significantly suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and proinflammatory cytokines mRNA levels. Taken together, these results suggest that the anti-inflammatory activity of isoeugenin is associated with the down-regulation of iNOS, COX-2, and pro-inflammatory cytokines in RAW264.7 cells. Accordingly, our results suggest that the new chromone isoegenin should be considered a potential treatment for inflammatory disease.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Chromones/pharmacology , Enzyme Inhibitors/pharmacology , Macrophages/drug effects , Nitric Oxide Synthase Type II/antagonists & inhibitors , Plant Extracts/pharmacology , Poaceae/chemistry , Rhizome/chemistry , Animals , Anti-Inflammatory Agents/isolation & purification , Chromones/chemistry , Cyclooxygenase 2/chemistry , Cytokines/genetics , Enzyme Inhibitors/isolation & purification , Lipopolysaccharides/pharmacology , Macrophages/cytology , Macrophages/metabolism , Mice , Nitric Oxide/metabolism , RAW 264.7 Cells , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
CONTEXT: Artemisia iwayomogi Kitamura (Compositae) has been very widely used for the treatment of acute or chronic hepatitis, jaundice, and gastritis. In the course of our continuing efforts to identify and quantify peroxynitrite scavengers from Compositae plants, A. iwayomogi was used in this study. OBJECTIVE: The present study was aimed to identify and quantify the peroxynitrite scavengers of A. iwayomogi. MATERIALS AND METHODS: Silica gel and ODS were used for column chromatography. The isolated compounds were quantified using an HPLC equipped with a Capcell Pak C18 column (5 µm, 250 mm × 4.6 mm i.d.), and the method was validated for the quality control. Peroxynitrite (ONOO(-))-scavenging activities of the compounds and extracts were evaluated on the measurement of highly fluorescent rhodamine 123 converted from non-fluorescent dihydrorhodamine (DHR)-123 under the presence of peroxynitrite. RESULTS: Based on the spectroscopic evidences, a new compound, 2"-O-caffeoylrutin (2"-O-trans-caffeic acid ester of quercetin 3-O-α-L-rhamnopyranosyl(1 â 6)-ß-D-glucopyranoside) was isolated and determined together with patuletin 3-O-glucoside, scopolin, scopoletin, rutin, 3,4-dicaffeoylquinic acid, and chlorogenic acid. All of them were potent peroxynitrite scavengers (IC50 ≤ 1.88 µg/mL). DISCUSSION AND CONCLUSION: The peroxynitrite scavengers were mainly distributed in the EtOAc fraction rather than the ether and BuOH fractions. The 70% MeOH extract exhibited a high peroxynitrite-scavenging activity. Through the validation, the present HPLC method was verified to be sufficiently sensitive, accurate, precise, and stable. Therefore, this method can be used for the quality control of A. iwayomogi.
Subject(s)
Artemisia , Free Radical Scavengers/isolation & purification , Peroxynitrous Acid/isolation & purification , Plant Components, Aerial , Plant Extracts/isolation & purification , Free Radical Scavengers/chemistry , Peroxynitrous Acid/chemistry , Plant Extracts/chemistryABSTRACT
The aim of the present study was to evaluate the comparative anti-inflammatory activities of Ixeris dentata (ID), Ixeris dentata var. albiflora (IDA), and Ixeris sonchifolia (IS) and to identify the main compounds present in extracts. The anti-inflammatory activity was evaluated through lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 murine macrophages. Five main compounds consisting of chlorogenic acid, caffeic acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, and luteolin were used for simultaneous high-performance liquid chromatography quantification. The total phenolic content present in ID (30 mg/g GAE), IDA (35.33 mg/g GAE), and IS (43.79 mg/g GAE) was correlated to the corresponding LPS-induced NO production inhibitory effect in RAW 264.7 cells as expressed with IC(50) values 26.19, 21.43, and 7.59 µg/mL, respectively. Luteolin 7-O-glucoside was found as the major compound in ID (8.76 mg/g dry weight) and IDA (10.35 mg/g dry weight) and luteolin 7-O-glucuronide was the major compound in IS (34.66 mg/g dry weight). Luteolin 7-O-glucoside and luteolin 7-O-glucuronide inhibited LPS-induced NO production with IC(50) values of 30 and 4.5 µM, respectively. Furthermore, luteolin, luteolin 7-O-glucoside, and luteolin 7-O-glucuronide suppressed the expression of iNOS and COX-2, and t-BHP-induced ROS generation in LPS-stimulated RAW 264.7 cells. These results clearly showed that the anti-inflammatory potential of ID, IDA, and IS extract are primarily due to their contents of luteolin 7-O-glucoside and luteolin 7-O-glucuronide, respectively.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Asteraceae/chemistry , Inflammation/drug therapy , Macrophages/drug effects , Phytotherapy , Plant Extracts/therapeutic use , Polyphenols/therapeutic use , Animals , Anti-Inflammatory Agents/analysis , Anti-Inflammatory Agents/pharmacology , Chromatography, High Pressure Liquid , Cyclooxygenase 2/metabolism , Inflammation/chemically induced , Lipopolysaccharides , Macrophages/metabolism , Mice , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/metabolism , Plant Extracts/chemistry , Plant Extracts/pharmacology , Polyphenols/analysis , Polyphenols/pharmacology , Reactive Oxygen Species/metabolismABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Rubus coreanus Miquel (Rosaceae), the Korean black raspberry, has traditionally been used to treat inflammatory diseases including diarrhea, asthma, stomach ailment, and cancer. Although previous studies showed that the 19α-hydroxyursane-type triterpenoids isolated from Rubus coreanus exerted anti-inflammatory activities, their effects on ulcerative colitis and mode of action have not been explored. This study was designed to assess the anti-inflammatory effects and the molecular mechanisms involving19α-hydroxyursane-type triterpenoid-rich fraction from Rubus coreanus (TFRC) on a mice model of colitis and lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. MATERIALS AND METHODS: Experimental colitis was induced by DSS for 7 days in ICR mice. Disease activity indices (DAI) took into account body weight, stool consistency, and gross bleeding. Histological changes and macrophage accumulation were observed by immunohistochemical analysis. Pro-inflammatory markers were determined using immunoassays, RT-PCR, and real time PCR. Signaling pathway involving nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) activation was determined by luciferase assay and Western blotting. RESULTS: In DSS-induced colitis mice, TFRC improved DAIs and pathological characteristics including colon shortening and colonic epithelium injury. TFRC suppressed tissue levels of pro-inflammatory cytokines and reduced macrophage infiltration into colonic tissues. In LPS-induced RAW 264.7 macrophages, TFRC inhibited the production of NO, PGE2, and pro-inflammatory cytokines by down-regulating the activation of NF-κB and p38 MAPK signaling. CONCLUSION: The study demonstrates that TFRC has potent anti-inflammatory effects on DSS-induced colonic injury and LPS-induced macrophage activation, and supports its possible therapeutic and preventive roles in colitis.
Subject(s)
Colitis/prevention & control , Dextran Sulfate/toxicity , Lipopolysaccharides/toxicity , Macrophages/drug effects , Plant Extracts/pharmacology , Rubus/chemistry , Triterpenes/analysis , Animals , Base Sequence , Cell Line , Colitis/chemically induced , Cytokines/biosynthesis , Cytokines/genetics , DNA Primers , Inflammation Mediators/metabolism , Macrophages/metabolism , Mice , Plant Extracts/chemistry , Polymerase Chain ReactionABSTRACT
BACKGROUND: Allium victorialis var. platyphyllum is an edible perennial herb and has been used as a vegetable or as a Korean traditional medicine. Allium species have received much attention owing to their diverse pharmacological properties, including antioxidative, anti-inflammatory, and anticancer activities. However, A. victorialis var. platyphyllum needs more study. METHODS: The chemopreventive potential of A. victorialis var. platyphyllum methanol extracts was examined by measuring 12-O-tetra-decanoylphorbol 13-acetate (TPA)-induced superoxide anion production in the differentiated HL-60 cells, TPA-induced mouse ear edema, and Ames/Salmonella mutagenicity. The apoptosis-inducing capabilities of the extracts were evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, 4',6-diamidino-2-phenylindole staining, and the DNA fragmentation assay in human colon cancer HT-29 cells. Antimetastatic activities of the extracts were also investigated in an experimental mouse lung metastasis model. RESULTS: The methanol extracts of A. victorialis var. platyphyllum rhizome (AVP-R) and A. victorialis var. platyphyllum stem (AVP-S) dose-dependently inhibited the TPA-induced generation of superoxide anion in HL-60 cells and TPA-induced ear edema in mice, as well as 7,12-dimethylbenz[a]anthracene (DMBA) and tert-butyl hydroperoxide (t-BOOH) -induced bacterial mutagenesis. AVP-R and AVP-S reduced cell viability in a dose-related manner and induced apoptotic morphological changes and internucleosomal DNA fragmentation in HT-29 cells. In the experimental mouse lung metastasis model, the formation of tumor nodules in lung tissue was significantly inhibited by the treatment of the extracts. CONCLUSIONS: AVP-R and AVP-S possess antioxidative, anti-inflammatory, antimutagenic, proapoptotic, and antimetastatic activities. Therefore, these extracts can serve as a beneficial supplement for the prevention and treatment of cancer.
ABSTRACT
The herbs of Euphorbia supina (Euphorbiaceae) have been used to treat hemorrhage, chronic bronchitis, hepatitis, jaundice, diarrhea, gastritis, and hemorrhoids as a medicinal herb. This work is aimed to qualitatively and quantitatively analyze the polyphenols with peroxynitrite-scavenging activities. The eight compounds: gallic acid, methyl gallate, avicularin, astragalin, juglanin, isoquercitrin 6â³-gallate, astragalin 6â³-gallate, and ellagic acid, were isolated from E. supina and used for HPLC analysis and peroxynitrite (ONOO(-))-scavenging assay. Simultaneous analysis of the eight compounds was performed on MeOH extract and its fractions. The contents in MeOH extract and peroxynitrite-scavenging activities of the dimer of gallic acid, ellagic acid (15.64 mg/g; IC50 0.89 µM), and two galloylated flavonoid glycosides, astragalin 6â³-gallate (13.72 mg/g; IC50 1.43 µM) and isoquercitrin 6â³-gallate (16.99 mg/g; IC50 1.75 µM), were high, compared to other compounds. The legendary uses of E. supina could be attributed to the high content of polyphenols, particularly ellagic acid, isoquercitrin 6â³-gallate, and astragalin 6â³-gallate as active principles.
Subject(s)
Drugs, Chinese Herbal/analysis , Ellagic Acid/analysis , Euphorbia , Flavonoids/analysis , Glycosides/analysis , Peroxynitrous Acid/metabolism , Drugs, Chinese Herbal/chemistry , Ellagic Acid/chemistry , Flavonoids/metabolism , Free Radical Scavengers/analysis , Free Radical Scavengers/metabolism , Glycosides/chemistryABSTRACT
The plant Polygonum aviculare L. (Polygonaceae) is an annual herbaceous plant which is known to be beneficial for treating gastroduodenal ulcer, hypertension, diarrhea, hemorrhage, and hemorrhoids. Ten phenolic compounds, including nine flavonoids (myricetin, quercetin, kaempferol, myricitrin, desmanthin-1, isoquercitrin, quercitrin, avicularin, juglanin), and gallic acid were used for simultaneous HPLC quantification and peroxynitrite-scavenging assay. Simultaneous quantification of these substances were performed on five extracts (EtOH-, MeOH-, 70% MeOH-, 30% MeOH-, and H2O extracts) as well as on the three fractions (Et2O-, EtOAc-, and BuOH fractions), under the condition of a Capcell Pak C18 column (5µm, 250mm×4.6mm i.d.) and a gradient elution of 0.05% trifluoroacetic acid (TFA) and MeOHCH3CN (60:40). Of the three fractions, the EtOAc fraction displayed the highest content of flavonoids (sum, 208.9mg/g) with the strongest peroxynitrite-scavenging activity (IC50, 2.68µg/mL). The activities of the eight compounds (myricitrin, isoquercitrin, avicularin, quercitrin, myricetin, desmanthin-1, quercetin, and kaempferol) were comparable to that of the positive control (l-penicillamine; IC50: 1.03µg/mL). These results suggest that folkloric medicinal uses of P. aviculare are mainly attributed to flavonoids, such as particularly highly contained myricetin, myricitrin, and desmanthin-1.
Subject(s)
Antioxidants/chemistry , Flavonoids/chemistry , Free Radical Scavengers/chemistry , Peroxynitrous Acid/chemistry , Polygonum/chemistry , Chromatography, High Pressure Liquid/methods , Phenols/chemistry , Plant Extracts/chemistryABSTRACT
Aster glehni (AG) is a Korean traditional herb that grows in Ulleungdo Island, Republic of Korea. None of the several reports on AG include a determination of the effect of AG on adipogenesis. The primary aim of this study was to determine whether AG attenuates adipogenesis in mouse 3T3-L1 cells and epididymal fat tissue. AG blocked the differentiation of 3T3-L1 preadipocytes in a concentration-dependent manner and suppressed the expression of adipogenesis-related genes such as PPAR γ , C/EBP α , and SREBP1c, the master regulators of adipogenesis. Male C57BL/6J mice were divided randomly and equally into 4 diet groups: control diet (CON), high-fat diet (HFD), HFD with 1% AG extract added (AG1), and HFD with 5% AG extract added (AG5). The experimental animals were fed HFD and the 2 combinations for 10 weeks. Mice fed HFD with AG gained less body weight and visceral fat-pad weight than did the mice fed HFD alone. Moreover, AG inhibited the expression of important adipogenic genes such as PPAR γ , C/EBP α , SREBP1c, LXR, and leptin in the epididymal adipose tissue of the mice treated with AG1 and AG5. These findings indicate antiadipogenic and antiobesity effects of AG and suggest its therapeutic potential in obesity and obesity-related diseases.
ABSTRACT
Traditionally, Hemistepta lyrata is consumed as a mountainous vegetable or a medicinal herb to treat inflammation, fever, hemorrhage, and hemorrhoids. In order to provide the scientific evidence of traditional uses of this plant, we identified and quantified thirteen active substances (caffeic acid, chlorogenic acid, and 3,5-di-O-caffeoylquinic acid as caffeoylquinic acids; apigenin, isorhoifolin, acacetin, linarin, diosmetin, diosmin, pectolinarigenin, and pectolinarin as flavones or their glycosides; kaempferol 3-O-rutinoside and rutin as flavonol glycosides) from H. lyrata and evaluated their peroxynitrite-scavenging activity. The chromatographic separation was performed on a Capcell Pak C18 column (5µm, 250mm×4.6mm i.d.) with a gradient elution of 0.05% TFA (trifluoroacetic acid) and 0.05% TFA in MeOH-CH(3)CN (60:40). Validation of HPLC methods on the linearity, LOD, LOQ, intra-day and inter-day variabilities, recovery, and repeatability proved that this method is selective, sensitive, precise, accurate, and reproducible. In peroxynitrite-scavenging assay, caffeic acid derivatives (chlorogenic acid, caffeic acid, and 3,5-di-O-caffeoylquinic acid) exhibited relatively lower IC(50) values than other substances tested. And HPLC simultaneous quantification showed that the 70% MeOH extract and the BuOH fraction contain a higher quantity of caffeic acid derivatives (17.82 and 30.09mg/g, consecutively). Therefore, caffeic acid derivatives could be the main contributors to the peroxynitrite-scavenging activity of H. lyrata than other phenolic substances.
Subject(s)
Asteraceae/chemistry , Chromatography, High Pressure Liquid/methods , Flavonoids/analysis , Quinic Acid/analogs & derivatives , Flavonoids/isolation & purification , Flavonoids/pharmacology , Inhibitory Concentration 50 , Limit of Detection , Medicine, Traditional , Peroxynitrous Acid/metabolism , Quinic Acid/analysis , Quinic Acid/isolation & purification , Quinic Acid/pharmacology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Animal experiments were performed to develop Salvia plebeia (Labiatae) as a medicinal herb with sedative and gastroprotective activities; the former activity was measured using a pentobarbital-induced assay and the latter activity was measured in two gastric lesion-induced assays (HCl/EtOH-induced and indomethacin/bethanechol-induced assays) in mice. The MeOH extract and its EtOAc fraction were effective, although the former was less active than the latter. Rosmarinic acid (RA) isolated from S. plebeia was active in the same method at 10 and 20 mg/kg (p.o.). HPLC quantification demonstrated that RA comprised the largest proportion (28.5% of the MeOH extract, 33.0% of EtOAc extract; 4.46% of dry weight) of S. plebeia. The contents of five other compounds were much less than that of RA, although the contents of the three glycosides, 6-hydroxyluteolin 7-O-glucoside (0.28% of dry weight), cynaroside (0.35%) and nepitrin (0.43%) were higher than those of the two aglycones, quercetin (0.024%) and eupatilin (0.058%). The HPLC method was validated in terms of linearity, precision, accuracy and reproducibility. These results suggest that the main polyphenol, RA, plays a major role in the sedative and gastroprotective effects of S. plebeia.
Subject(s)
Chromatography, High Pressure Liquid/methods , Plant Extracts/pharmacology , Polyphenols/pharmacology , Salvia/chemistry , Animals , Anti-Ulcer Agents/administration & dosage , Anti-Ulcer Agents/isolation & purification , Anti-Ulcer Agents/pharmacology , Disease Models, Animal , Dose-Response Relationship, Drug , Hypnotics and Sedatives/administration & dosage , Hypnotics and Sedatives/isolation & purification , Hypnotics and Sedatives/pharmacology , Male , Mice , Mice, Inbred ICR , Peptic Ulcer/drug therapy , Peptic Ulcer/pathology , Plant Extracts/administration & dosage , Plant Extracts/chemistry , Polyphenols/isolation & purification , Reproducibility of ResultsABSTRACT
On high performance liquid chromatography, the caffeoylquinic acid (CQ) occupying the highest proportion of the water-ethanol (7:3) extract of Aster glehni (Compositae) leaves was 3-Op-coumaroylquinic acid (46.10 ± 4.22 mg/g of dried weight) among CQs tested. The IC50 of the water-ethanol (7:3) extract was 4.23 ± 0.24 µg/mL in the peroxynitrite (ONOO(-))-scavenging assay. Phytochemical isolation from A. glehni extract yielded three kaempferol glycosides. The water-ethanol (7:3) extract and both p-coumaric acid and caffeic acid, phenylpropanoid moieties of CQs, had sedative effects in pentobarbital-induced sleeping time in mice and anticonvulsant effects in pentylenetetrazole-induced mice. Furthermore, the phenolic substance-rich W-E (7:3) extract of A. glehni could be used to treat anxiety or convulsion partly due to its peroxynitrite-scavenging mechanism.
Subject(s)
Anticonvulsants/pharmacology , Free Radical Scavengers/pharmacology , Hypnotics and Sedatives/pharmacology , Phenols/pharmacology , Plant Extracts/pharmacology , Seizures/prevention & control , Sleep/drug effects , Animals , Anticonvulsants/chemistry , Anticonvulsants/isolation & purification , Aster Plant/chemistry , Chromatography, High Pressure Liquid , Disease Models, Animal , Ethanol/chemistry , Free Radical Scavengers/chemistry , Free Radical Scavengers/isolation & purification , Hypnotics and Sedatives/chemistry , Hypnotics and Sedatives/isolation & purification , Kaempferols/isolation & purification , Kaempferols/pharmacology , Male , Mice , Mice, Inbred ICR , Pentobarbital/pharmacology , Pentylenetetrazole , Peroxynitrous Acid/chemistry , Phenols/chemistry , Phenols/isolation & purification , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Leaves , Quinic Acid/analogs & derivatives , Quinic Acid/isolation & purification , Quinic Acid/pharmacology , Seizures/chemically induced , Solvents/chemistry , Time Factors , Water/chemistryABSTRACT
As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we have evaluated the anti-inflammatory effects of euscaphic acid (19α-hydroxyursane-type triterpenoids, EA) isolated from roots of Rosa rugosa and its underlying molecular mechanisms in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. EA concentration-dependently reduced the production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß) induced by LPS in RAW 264.7 macgophages. Consistent with these data, expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein and iNOS, COX-2, TNF-α, and IL-1ß mRNA were inhibited by EA in a concentration-dependent manner. In addition, EA attenuated LPS-induced DNA binding and transcriptional activity of nuclear factor-kappa B (NF-κB), which was accompanied by a parallel reduction of degradation and phosphorylation of inhibitory kappa Bα (IκBα) and consequently by decreased nuclear translocation of p65 subunit of NF-κB. Pretreatment with EA significantly inhibited the LPS-induced phosphorylation of IκB kinase ß (IKKß), p38, and JNK, whereas the phosphorylation of ERK1/2 was unaffected. Furthermore, EA interfered with the LPS-induced clustering of TNF receptor-associated factor 6 (TRAF6) with interleukin receptor associated kinase 1 (IRAK1) and transforming growth factor-ß-activated kinase 1 (TAK1). Taken together, these results suggest that EA inhibits LPS-induced inflammatory responses by interference with the clustering of TRAF6 with IRAK1 and TAK1, resulting in blocking the activation of IKK and MAPKs signal transduction to downregulate NF-κB activations.
Subject(s)
Inflammation/drug therapy , Macrophages/immunology , NF-kappa B/metabolism , Toll-Like Receptor 4/metabolism , Triterpenes/pharmacology , Animals , Cell Line , Cyclooxygenase 2/biosynthesis , Dinoprostone/biosynthesis , Extracellular Signal-Regulated MAP Kinases/metabolism , I-kappa B Proteins/metabolism , Interleukin-1 Receptor-Associated Kinases/metabolism , Interleukin-1beta/biosynthesis , JNK Mitogen-Activated Protein Kinases/metabolism , Lipopolysaccharides/immunology , MAP Kinase Kinase Kinases/metabolism , MAP Kinase Signaling System/drug effects , Macrophages/drug effects , Macrophages/metabolism , Mice , NF-KappaB Inhibitor alpha , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type II/biosynthesis , Phosphorylation/drug effects , Phytotherapy , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rosa/chemistry , TNF Receptor-Associated Factor 6/drug effects , TNF Receptor-Associated Factor 6/metabolism , Transcription Factor RelA/metabolism , Transcription, Genetic/drug effects , Tumor Necrosis Factor-alpha/biosynthesis , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
The leaves of Saussurea grandifolia (Compositae) are used as chwinamul, a well-known Korean mountainous vegetable, or to treat hepatitis and hematuria. Since the methanolic extract of S. grandifolia leaves exhibit a potent peroxynitrite-scavenging effect, phytochemical and high-performance liquid chromatographic (HPLC) analyses were employed to identify and simultaneously quantify the active components: chlorogenic acid and 3,5-di-O-caffeoylquinic acid (3,5-DQ) as caffeoylquinic acids, and quercetin, isoquercitrin (quercetin-3-glucoside), saxifragin (quercetin-5-glucoside), and rutin (quercetin-3-rutinoside). Validation of HPLC methods was performed to verify the linearity, LOD, LOQ, intra-day and inter-day variabilities, recovery, and repeatability to ensure that this method is selective, sensitive, precise, accurate and reproducible. In particular, leaves contained saxifragin with potent peroxynitrite-scavenging activity (IC(50), 0.67 µM) as 4.65 mg/g dry weight, which is equivalent to 33.74 mg/g extract.
Subject(s)
Chemistry, Pharmaceutical/standards , Free Radical Scavengers/analysis , Peroxynitrous Acid/analysis , Plant Extracts/analysis , Quercetin/analysis , Saussurea/chemistry , Chemistry, Pharmaceutical/methods , Chromatography, High Pressure Liquid/methods , Chromatography, High Pressure Liquid/standards , Free Radical Scavengers/chemistry , Glycosides/analysis , Peroxynitrous Acid/chemistry , Plant Leaves/chemistry , Reproducibility of ResultsABSTRACT
To identify an analgesic/anti-inflammatory component from the leaves of Rhododendron aureum (Ericaceae), phytochemical isolation and pharmacological assays (writhing assays and vascular permeability assay for analgesic action in mice; carrageenan-induced paw edemaand TPA-induced ear edema assays of anti-inflammatory action in rats) were performed. Four compounds were isolated from the active fraction (BuOH fraction) by silica gel column chromatography and identified as (-)-rhododendrol, (-)-rhododendrin, avicularin and hyperoside by spectroscopic methods. Rhododendrin, the main compound of the BuOH fraction, exhibited significant analgesic actions in mice and anti-inflammatory actions in rats. This compound accounted for 3.1% of the MeOH extract and 0.48% of dried leaves, respectively, on HPLC analysis. These results suggest that rhododendrin is the major biologically active substance in the leaves of R. aureum with analgesic/anti-inflammatory activity.
Subject(s)
Analgesics/pharmacology , Anti-Inflammatory Agents/pharmacology , Plant Extracts/pharmacology , Rhododendron/chemistry , Analgesics/isolation & purification , Animals , Anti-Inflammatory Agents/isolation & purification , Chromatography, High Pressure Liquid , Disease Models, Animal , Glycosides/isolation & purification , Glycosides/pharmacology , Guinea Pigs , Inflammation/drug therapy , Male , Medicine, Traditional , Mice , Mice, Inbred ICR , Pain/drug therapy , Pain Measurement , Plant Leaves , Rats , Rats, Sprague-DawleyABSTRACT
The unripe fruits of Rubus coreanus (Rosaceae) are used in traditional Chinese medicine to relieve kidney dysfunction. In the present study, we evaluated the protective effects of the triterpenoid glycoside niga-ichigoside F1 (NIF1) and of its aglycone 23-hydroxytormentic acid (23-HTA) isolated from the unripe fruits of Rubus coreanus (Rosaceae) against cisplatin-induced cytotoxicity in renal epithelial LLC-PK1 cells. Pretreating LLC-PK1 cells with 23-HTA or NIF1 was found to prevent cisplatin-induced cytotoxicity and apoptosis. In addition, 23-HTA or NIF1 pretreatment significantly improved the changes associated with cisplatin toxicity by increasing levels of glutathione (GSH) and decreasing levels of malondialdehyde (MDA) and reactive oxygen species (ROS). The activity of antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) was significantly lower in cisplatin-treated LL-PK1 cells, and 23-HTA or NIF1 treatment notably increased the these enzyme activity and protein and mRNA levels of CAT and manganese SOD (MnSOD). Moreover, cisplatin caused a significant decrease in nuclear levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and pretreatment with 23-HTA or NIF1 significantly suppressed the cisplatin-induced translocation of Nrf2 in LLC-PK1 cells. Taken together, these results suggest that 23-HTA ameliorates cisplatin-induced toxicity via modulation of antioxidant enzymes through activation of Nrf2 in LLC-PK1 cells.
Subject(s)
Antineoplastic Agents/toxicity , Antioxidants/pharmacology , Cisplatin/toxicity , Glycosides/pharmacology , Kidney/drug effects , Oxidative Stress/drug effects , Triterpenes/pharmacology , Animals , Apoptosis/drug effects , Cell Line , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Cell Survival/drug effects , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Fruit/chemistry , Fruit/growth & development , Gene Expression Regulation, Enzymologic/drug effects , Glutathione/metabolism , Kidney/metabolism , NF-E2-Related Factor 2/metabolism , Oxidoreductases/genetics , Oxidoreductases/metabolism , Protein Transport/drug effects , RNA, Messenger/metabolism , Rosaceae/chemistry , Sus scrofaABSTRACT
The aerial part of Rubus rigidus var. camerunensis (Rosaceae) is used to treat respiratory and cardiovascular disorders in the Cameroonian traditional medicine. The ethanol extract exhibited more potent antioxidant activity (E(max)s of 119% and 229% activity on DPPH and ß-carotene test) than aqueous extract. Bioactivity-guided fractionation of the ethanol extract based on free radical-scavenging assay (DPPH assay) afforded five flavonoid glycosides (four flavonol glycosides and an anthocyanin) and three glucosides of 19α-hydroxyursane-type triterpenoid (two monomeric and one dimeric triterpenoids). The flavonoids were identified as kaempferol 3-O-(2â³-O-E-p-coumaroyl)-ß-D-glucopyranoside (1), kaempferol-3-O-ß-D-glucopyranoside (astragalin, 2), kaempferol-3-O-α-L-arabinofuranoside (juglanin, 3), quercetin-3-O-ß-D-glucopyranoside (isoquercitrin, 4), pelargonidin-3-O-ß-D-glucopyranoside (callistephin, 5). The three triterpenoids were 2α, 3ß, 19α, 23-tetrahydroxyurs-12-ene-28-O-ß-D-glucopyranosyl ester (nigaichigoside F(1), 6), 2α, 3ß, 19α-trihydroxyurs-12-ene-23-carboxyl-28-O-ß-D-glucopyranosyl ester (suavissimoside R(1), 7) as monomeric triterpenoids and coreanoside F(1) (8) as a dimeric triterpenoid. The flavonoids exhibited potent antioxidant activities (66 to 93.56% against DPPH radical) and they were also active on ß-carotene test. Coreanoside F(1) exhibited a 63% antioxidant activity, meanwhile the other two triterpenoids showed a weak activity. Three important facts on structure-activity relationship were observed: Compound 8, a dimeric triterpenoid glycoside, strongly enhanced antioxidant activity of its monomers, compound 3 with 3-O-α-L-arabinofuranyl has much more potent activity than compound 2 with 3-O-ß-D-glucopyranosyl, and antocyanin (5) is more potent than its corresponding flavonol glycosides.
Subject(s)
Flavonoids/isolation & purification , Free Radical Scavengers/isolation & purification , Glycosides/isolation & purification , Plant Extracts/isolation & purification , Rosaceae/chemistry , Triterpenes/isolation & purification , Biphenyl Compounds/chemistry , Dimerization , Flavonoids/chemistry , Flavonoids/pharmacology , Free Radical Scavengers/chemistry , Free Radical Scavengers/pharmacology , Free Radicals/chemistry , Glycosides/chemistry , Glycosides/pharmacology , Molecular Structure , Picrates/chemistry , Plant Components, Aerial/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Triterpenes/chemistry , Triterpenes/pharmacologyABSTRACT
Endometriosis, a disease affecting 5-15% of women of reproductive age, is characterized by the ectopic growth of endometrial tissue. Costunolide, a sesquiterpene lactone, has anti-proliferative and pro-apoptotic activities that may be efficacious in therapy for endometriosis. In the present study, we investigated the effect of costunolide on the cell growth and apoptosis of endometriotic cells. We found that costunolide significantly inhibited the cell viability of 11Z and 12Z human endometriotic epithelial cells. Interestingly, endometriotic cells were more sensitive to costunolide treatment than immortalized endometrial cells (HES). Costunolide induced apoptosis of 11Z cells in a time-dependent manner as shown by accumulation of sub-G1 population. In addition, treatment with costunolide induced the activation of caspase-3, -8, and -9 in a dose- and time-dependent manner. Pretreatment with the broad caspase inhibitor z-VAD-fmk significantly reversed the costunolide-induced inhibition of cell viability in 11Z cells. We further demonstrated that costunolide inhibited the activation of Akt and nuclear factor kappa B (NFκB) and the expression of anti-apoptotic factors B-cell lymphoma-extra lage (Bcl-xL) and X-linked inhibitor of apoptosis protein (XIAP) in 11Z cells. These results suggest that costunolide induces apoptosis in human endometriotic epithelial cells by inhibiting the prosurvival NFκB and Akt pathway, leading to the downregulation of anti-apoptotic protein Bcl-xL and XIAP and the activation of caspases.