Anti-Inflammatory Effect of Artemisia argyi on Ethanol-Induced Gastric Ulcer: Analytical, In Vitro and In Vivo Studies for the Identification of Action Mechanism and Active Compounds.

Artemisia argyi is widely used as traditional medicine in East Asia. However, its effects against inflammation and gastric ulcers have not been reported yet. We analyzed anti-inflammatory activity and its molecular mechanisms of A. argyi using RAW264.7 cells line, then evaluated the curative efficacy in rats with acute gastric ulcers. Nitric oxide and IL-6 production was measured using Griess reagent and an ELISA kit. Inducible nitric oxide synthase (iNOS), interleukin (IL)-6, and mucin (MUC)1, MUC5AC, and MUC6 mRNA were determined by SYBR Green or Taqman qRT-PCR methods. The phosphorylation of ERK, JNK, p38, and c-Jun protein were detected by western blotting. RW0117 inhibited LPS-induced NO and IL-6 production. The mRNA levels of iNOS and IL-6 were strongly suppressed. The phosphorylation of ERK, JNK, and c-Jun decreased by treatment with RW0117. Oral administration of RW0117 recovered the amount of mucin mRNA and protein level that was decreased due to gastric ulcers by HCl-EtOH. A. argyi exhibited strong anti-inflammatory effects and contributed to the modulation of HCl-EtOH-induced gastric ulcer in rats.

Chemical constituents of the root bark of Ulmus davidiana var. japonica and their potential biological activities.

The root bark of Ulmus davidiana var. japonica (Ulmaceae), commonly known as yugeunpi, has been used as a traditional Korean medicine for the treatment of gastroenteric and inflammatory disorders. As part of continuing projects to discover bioactive natural products from traditional medicinal plants with pharmacological potential, phytochemical investigation of the root bark of this plant was carried out. This led to the successful isolation of a new chromane derivative (1) and 22 known compounds: catechin derivatives (2-5), megastigmane glycoside (6), dihydrochalcone glycosides (7 and 8), flavanone glycosides (9 and 10), coumarins (11 and 12), lignan derivatives (13-17), and phenolic compounds (18-23). The structure of the new compound (1) was determined with 1D and 2D NMR spectroscopy and HR-ESIMS, and its absolute configurations were achieved by chemical reactions and the gauge-including atomic orbital (GIAO) NMR chemical shifts calculations. All the isolated compounds were evaluated for their potential biological activities including neuro-protective, anti-neuroinflammatory, and anti-Helicobacter pylori activities. Among the isolates, compounds 1, 8, and 20 displayed stronger potency by causing a greater increase in the production and the activity of nerve growth factor (NGF) in C6 glioma cells (147.04 ±â€¯4.87, 206.27 ±â€¯6.70, and 143.70 ±â€¯0.88%, respectively), whereas compounds 11, 14, and 19 inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated murine microglial cells (IC50 of 18.72, 12.31, and, 21.40 µM, respectively). In addition, compounds 1, 11, 18, and 20 showed anti-H. pylori activity with MIC values of 25 or 50 µM against two strains of H. pylori 51 and 43504. These findings provide scientific evidence that supports the traditional usage of U. davidiana var. japonica root bark in the treatment of gastroenteric and inflammatory disorders.

Bioactivity evaluations of betulin identified from the bark of Betula platyphylla var. japonica for cancer therapy.

Identification of bioactive natural products with anticancer activity as well as alleviating effects on chemotherapy-induced side effects has significant implications for cancer treatment. Betula platyphylla var. japonica, commonly known as Asian white birch, has been used in Chinese traditional medicine for a variety of purposes. In this study, the medicinal properties of betulin from B. platyphylla var. japonica useful for cancer management were investigated. LC/MS analysis revealed that betulin is a main chemical component of the EtOH extract of B. platyphylla var. japonica bark, and betulin was isolated from EtOH extract using an LC/MS-guided isolation method. Its structure was identified with 1H and 13C NMR spectroscopic data and LC/MS analysis and then compared to the previously reported spectroscopic and physical data. We first verified the cytotoxicity of betulin against three human lung adenocarcinoma cell lines, A549, H1264, and Calu-6, with IC50 values ranging from 18.7 to 39.6 µM. Regarding alleviation of side effects associated with anticancer chemotherapy, betulin ameliorated cisplatin-induced renal cell damage to 80% of the control value from the concentration of 5 µM. In addition, betulin showed anti-gastritis activity against ethanol-induced gastric damage in rats and notably reduced the gastric damage index compared to control in a concentration-dependent manner. These findings provide the first experimental evidence for potential use of B. platyphylla var. japonica as a functional food for cancer treatment that simultaneously alleviates the side effects of chemotherapy.

Optimization of Extraction Conditions for Phenolic Acids from the Leaves of Melissa officinalis L. Using Response Surface Methodology.

BACKGROUND: Melissa officinalis L. is a well-known medicinal plant from the family Lamiaceae, which is distributed throughout Eastern Mediterranean region and Western Asia. OBJECTIVE: In this study, response surface methodology (RSM) was utilized to optimize the extraction conditions for bioactive compounds from the leaves of M. officinalis L. MATERIALS AND METHODS: A Box-Behnken design (BBD) was utilized to evaluate the effects of three independent variables, namely extraction temperature (°C), methanol concentration (%), and solvent-to-material ratio (mL/g) on the responses of the contents of caffeic acid and rosmarinic acid. RESULTS: Regression analysis showed a good fit of the experimental data. The optimal condition was obtained at extraction temperature 80.53°C, methanol concentration 29.89%, and solvent-to-material ratio 30 mL/g. CONCLUSION: These results indicate the suitability of the model employed and the successful application of RSM in optimizing the extraction conditions. This study may be useful for standardizing production quality, including improving the efficiency of large-scale extraction systems. SUMMARY: The optimum conditions for the extraction of major phenolic acids from the leaves of Melissa officinalis L. were determined using response surface methodologyBox-Behnken design was utilized to evaluate the effects of three independent variablesQuadratic polynomial model provided a satisfactory description of the experimental dataThe optimized condition for simultaneous maximum contents of caffeic acid and rosmarinic acid was determined. Abbreviations used: RSM: Response surface methodology, BBD: Box-Behnken design, CA: Caffeic acid, RA: Rosmarinic acid, HPLC: High-performance liquid chromatography.

In vitro assessment of selected Korean plants for antioxidant and antiacetylcholinesterase activities.

CONTEXT: Antiacetylcholinesterase (AChE) drugs have been a main therapeutic treatment for Alzheimer's disease because increased AChE levels play a key role in reducing neurotransmission. OBJECTIVES: Extracts from 35 Korean plants were selected and screened for antioxidant and anti-cholinesterase activity to explore new sources derived from Korean natural resources that could be used as AD therapeutic agents. MATERIALS AND METHODS: The antioxidant effect of extracts from 35 selected Korean plants was determined using two most common free radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS). Additionally, the effect of extracts, identified as antioxidants, on acetylcholinesterase inhibition was assessed by an acetylcholinesterase assay kit. RESULTS: Out of 36 extracts of 35 plants tested, Oenothera biennis L. (9.09 µg/mL), Saururus chinensis (Lour.) Baill. (9.52 µg/mL) and Betula platyphylla var. japonica (9.85 µg/mL) showed strong DPPH scavenging activity. Twelve other extracts also exerted moderate free radical scavenging activities with IC50 values ranging from 10 to 50 µg/mL. Antioxidant capacity detected by ABTS assay was only significant in O. biennis (23.40 µg/mL), while the other extracts were weak or unable to reduce the production of ABTS. Based on the antioxidant activities of these plant extracts, 19 extracts with IC50 values less than 100 µg/mL in DPPH assay were selected for further AChE inhibition assay. Among the extracts tested, the IC50 value for Prunella vulgaris var. lilacina NAKAI (18.83 µg/mL) in AChE inhibitory activity was the lowest, followed by O. biennis (20.09 µg/mL) and Pharbitis nil Chosy (22.79 µg/mL). CONCLUSIONS: Considering complex multifactorial etiology of AD, the extracts of P. vulgaris var. lilacina (aerial part), O. biennis (seed) and P. nil (seed) may be safe and ideal candidates for future AD modifying therapies.

Triterpene saponins from Pleurospermum kamtschaticum and their biological activity.

Eleven new triterpene saponins (1-11), together with fourteen known triterpene and triterpene saponins (12-25) were isolated from a MeOH extract of Pleurospermum kamtschaticum HOFFMANN (Umbelliferae). The chemical structures of the new compounds (1-11) were determined by means of MS, (1)H-NMR, (13)C-NMR, correlated spectroscopy (COSY), heteronuclear multiple bond correlation (HMBC), total correlated spectroscopy (TOCSY) and nuclear Overhauser effect spectroscopy (NOESY) to be pleurosaponin A (1)-K (11). The isolated compounds were tested for their cytotoxicity against four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, HCT15) in vitro using the sulforhodamine B bioassay (SRB) assay. All compounds showed little cytotoxicity against tested cell lines (IC(50) >30 µM).

A new flavonol glycoside from Hylomecon vernalis.

Purification of a MeOH extract from the aerial parts of Hylomecon vernalis Maxim. (Papaveraceae) using column chromatography furnished a new acetylated flavonol glycoside (1), together with twenty known phenolic compounds (2-21). Structural elucidation of 1 was based on 1D- and 2D-NMR spectroscopy data analysis to be quercetin 3-O-[4‴-O-acetyl-α-L-arabinopyranosyl]-(1‴→6″)-ß-D-galactopyranoside (1). The structures of compounds 2-21 were elucidated by spectroscopy and confirmed by comparison with reported data; quercetin 3-O-[2‴-O-acetyl-α-L-arabinopyranosyl]-(1‴→6″)-ß -D-galactopyranoside (2), quercetin 3-O-α-L-arabinopyranosyl-(1‴→6″)-ß-D-galactopyranoside (3), quercetin 3-O-ß -D-galactopyranoside (4), kaempferol 3,7-O-α-L-dirhamnopyranoside (5), diosmetin 7-O-ß -D-glucopyranoside (6), diosmetin 7-O-ß -D-xylopyranosyl-(1‴→6″)-ß-D-glucopyranoside (7), p-hydroxybenzoic acid (8), protocatechuic acid (9), caffeic acid (10), 6-hydroxy-3,4-dihydro-1-oxo-ß -carboline (11), (Z)-3-hexenyl-ß -D-glucopyranoside (12), (E)-2-hexenyl-ß -D-glucopyranoside (13), (Z)-3-hexenyl-α-Larabinopyranosyl-(1″→6')-ß-D-glucopyranoside (14), oct-1-en-3-yl-α-L-arabinopyranosyl-(1″→6')-ß-D-glucopyranoside (15), benzyl-ß-D-apiofuranosyl-(1″→6')-ß-D-glucopyranoside (16), benzyl-α-L-arabinopyranosyl-(1″→6')-ß-D-glucopyranoside (17), benzyl-ß-D-xylopyranosyl-(1″→6')-ß-Dglucopyranoside (18), 2-phenylethyl-α-L-arabinopyranosyl-(1″→6')-ß-D-glucopyranoside (19), 2-phenylethyl-ß-D-apiofuranosyl-(1″→6')-ß-D-glucopyranoside (20), and aryl-ß-D-glucopyranoside (21). Compounds 2-21 were isolated for the first time from this plant. The isolated compounds were tested for cytotoxicity against four human tumor cell lines in vitro using a Sulforhodamin B bioassay.

Phenolic constituents of Acorus gramineus.

The purification of a MeOH extract from the rhizome of Acorus gramineus (Araceae) using column chromatography furnished two new stereoisomers of phenylpropanoid, acoraminol A (1) and acoraimol B (2). It also furnished 17 known phenolic compounds, ß-asarone (3), asaraldehyde (4), isoacoramone (5), propioveratrone (6), (1'R,2'S)-1',2'-dihydroxyasarone (7), (1'S,2'S)-1',2'-dihydroxyasarone (8), 3',4'-dimethoxycinnamyl alcohol (9), 3',4',5'-trimethoxycinnamyl alcohol (10), kaempferol 3-methyl ether (11), 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-1,3-propanediol (12), hydroxytyrosol (13), tyrosol (14), (2S,5S)-diveratryl-(3R,4S)-dimethyltetrahydrofuran (15), (7S,8R)-dihydrodehydrodiconiferyl alcohol (16), 7S,8S-threo-4,7,9,9'-tetrahydroxy-3,3'-dimethoxy-8-O-4'-neolignan (17), 7S,8R-erythro-4,7,9,9'-tetrahydroxy-3,3'-dimethoxy-8-O-4'-neolignan (18), and dihydroyashsbushiketol (19). The structures of the new compounds were elucidated by analysis of spectroscopic data including 1D and 2D NMR data. The absolute configurations of 1 and 2 were determined using the convenient Mosher ester procedure. Compounds 5-19 were isolated for the first time from this plant source. The isolated compounds were tested for cytotoxicity against four human tumor cell lines in vitro using a Sulforhodamine B (SRB) bioassay.

Three new megastigmane glucopyranosides from the Cardamine komarovii.

Three new megastigmane glucopyranosides, komaroveside A [(3S,4R,5Z,7E)-3,4-dihydroxy-5,7-megastigmadien-9-one-3-O-ß-D-glucopyranoside] (1), komaroveside B [(3S,4S,5S,6R,7E)-5,6-epoxy-3,4-dihydroxy-7-megastigmen-9-one-3-O-ß-D-glucopyranoside] (2) and komaroveside C [(3S,4S,5S,6R,7E,9S)-5,6-epoxy-3,4,9-trihydroxy-7-megastigmen-3-O-ß-D-glucopyranoside] (3) were isolated, together with eight known compounds, from Cardamine komarovii. The identification of these compounds and the elucidation of their structures were based on 1D- and 2D-NMR spectral data analysis. The isolated compounds were tested for their cytotoxicity against four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, HCT15) in vitro using the sulforhodamine B bioassay.

New triterpenoids from the tubers of Corydalis ternata: structural elucidation and bioactivity evaluation.

Two new triterpene glycosides, coryternic acid 3-O-ß-D-glucuronopyranoside (1) and coryternic acid 3-O-ß-D-glucuronopyranoside-6'-O-methyl ester (2), were isolated from a MeOH extract of the tubers of Corydalis ternata. Acidic hydrolysis of 1 and 2 yielded a new triterpene as their aglycone, coryternic acid (3). The structures of these new compounds were determined through spectral analysis, including extensive 2D-NMR data. In this study we reported that triterpenoids were first isolated from the genus Corydalis. Compound 2 exhibited significant cytotoxicity against the A549, SK-OV-3, SK-MEL-2, and HCT-15 cell lines (IC50 = 15.16, 17.07, 13.32, and 11.95 µM, respectively) and significantly reduced NO production in lipopolysaccharide (LPS)-activated microglia/BV-2 cells with an IC50 value of 16.2 µM without cell toxicity.

Cytoprotective and anti-inflammatory effects of spinasterol via the induction of heme oxygenase-1 in murine hippocampal and microglial cell lines.

Spinasterol, which is isolated from the aerial parts of Aster scaber Thunb. (Asteraceae), is involved in various biological activities. In this study, we report the efficacy of spinasterol in effectively modulating the regulation of antioxidative and anti-inflammatory activity through the upregulation of heme oxygenase (HO)-1 in murine hippocampal HT22 cells and BV2 microglia. We showed that spinasterol increased the cellular resistance of HT22 cells to oxidative injury caused by the glutamate-induced cytotoxicity by extracellular signal-regulated kinase (ERK) pathway-dependent expression of HO-1. Furthermore, spinasterol suppressed the lipopolysaccharide (LPS)-induced expression of pro-inflammatory enzymes and inflammatory mediators in BV2 microglia. Spinasterol also suppressed the production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß) through extracellular signal-regulated kinase (ERK) pathway-dependent expression of HO-1. These results suggest that spinasterol has a therapeutic potential against neurodegenerative diseases that are caused by oxidative stress and neuroinflammation.

Benzylisoquinoline alkaloids from the tubers of Corydalis ternata and their cytotoxicity.

Chemical investigation of the tubers of Corydalis ternata resulted in the isolation and characterization of four new benzylisoquinoline alkaloids, epi-coryximine (1) and coryternatines A-C (2-4), along with 10 known alkaloids (5-14). Their structures were established on the basis of extensive spectroscopic data analyses and comparison with spectroscopic data reported. In addition, the cytotoxicities of the alkaloids (1-14) were evaluated by determining their inhibitory effects on several human tumor cell lines (A549, SK-OV-3, SK-MEL-2, and HCT-15) using the SRB assay. Compound 8 showed significant cytotoxicity against A549, SK-OV-3, SK-MEL-2, and HCT-15 cell lines (IC(50)=8.34, 5.14, 7.87, and 2.86 microM, respectively). The four new compounds (1-4) exhibited selective cytotoxicity against the HCT-15 cell line.

The ameliorating effect of the extract of the flower of Prunella vulgaris var. lilacina on drug-induced memory impairments in mice.

Prunella vulgaris var. lilacina is widely distributed in Korea, Japan, China, and Europe, and its flowers are used to treat inflammation in traditional Chinese medicine. In the present study, we studied the effects of the ethanolic extract of the flower of P. vulgaris var. lilacina (EEPV) on drug-induced learning and memory impairment using the passive avoidance, the Y-maze, and the Morris water maze tasks in mice. EEPV (25 or 50 mg/kg, p.o.) significantly ameliorated scopolamine-induced cognitive impairments in the passive avoidance and Y-maze tasks (P<0.05). In the Morris water maze task, EEPV (25 mg/kg, p.o.) significantly shortened escape latencies in training-trials. Furthermore, swimming times within the target zone during the probe-trial were significantly increased as compared with scopolamine-treated mice (P<0.05). In addition, the reduced latency induced by MK-801 treatment in the passive avoidance task was ameliorated by EEPV (25 mg/kg, p.o.) (P<0.05). Additionally, the ameliorating effect of EEPV on scopolamine-induced memory dysfunction was antagonized by a sub-effective dose of MK-801. These results suggest that EEPV would be useful for treating cognitive impairments induced by cholinergic dysfunction, and that it exerts its effects via NMDA receptor signaling.

New benzamide derivatives and NO production inhibitory compounds from Limonia acidissima.

Three new benzamide derivatives, N-{[ P-(3,7-dimethyl-6 R,7-dihydroxy-4 R-octadecanoyloxy-2-octenyloxy)phenyl]ethyl} benzamide (1), N-{[ P-(3,7-dimethyl-6 R,7-dihydroxy-4 R-9'''( E)-octadecenoyloxy-2-octenyloxy)phenyl]ethyl} benzamide (2), and N-{[ P-(3,7-dimethyl-6 R,7-epoxy-4 R-9'''( E)-octadecenoyloxy-2-octenyloxy)phenyl]ethyl} benzamide (3), together with 10 known compounds (4-13), were isolated from the bark of Limonia acidissima. The structures of these new compounds were determined through spectral analyses, including extensive 2D NMR data. Among the isolates, 13 alpha,14 beta,17 alpha-lanosta-7,9,24-triene-3 beta,16 alpha-diol (8), 4-methoxy-1-methyl-2(1 H)-quinolinone (10), and 13 alpha,14 beta,17 alpha-lanosta-7,24-diene-3 beta,11 beta,16 alpha-triol (13) potently inhibited nitric oxide (NO) production in microglia cells.

Triterpenoic acids of Prunella vulgaris var. lilacina and their cytotoxic activities in vitro.

The column chromatographic separation of the MeOH extract from the aerial parts of Prunella vulgaris var. lilacina Nakai led to the isolation of fifteen triterpenoic acids (2-6, 9-13, 16-20), four flavonoids (14, 21-23), four phenolics (7, 8, 15, 24), and a diterpene (1). Their structures were determined by spectroscopic methods to be trans-phytol (1), oleanic acid (2) ursolic acid (3), 2alpha,3alpha,19alpha-trihydroxyurs-12en-28oic acid (4), 2alpha,3alpha-dihydroxyurs-12en-28oic acid (5), maslinic acid (6), caffeic acid (7), phydroxy cinnamic acid (8), 2alpha,3alpha,19alpha,23-tetrahydroxyurs-12en-28oic acid (9), 2alpha,3alpha,23-trihydroxyurs-12en-28oic acid (10), 2alpha,3beta-dihydroxyurs-12en-28oic acid (11), 2alpha,3beta,24-trihydroxyolea-12en-28oic acid (12), (12R, 13S)-2alpha,3alpha,24,trihydroxy-12,13-cyclo-taraxer-14-en-28oic acid (13), quercertin 3-O-beta-D-glucopyranoside (14), rosmarinic acid (15), 2alpha,3alpha,24-trihydroxyurs-12,20(30)-dien-28oic acid (16), 2alpha,3alpha,24-trihydroxyolea-12en-28oic acid (17), 2alpha,3beta,19alpha,24-tetrahydroxyurs-12en-28oic acid 28-O-Dglucopyranoside (18), 2alpha,3alpha,19alpha,24-tetrahydroxyurs-12en-28oic acid 28-O-D-glucopyranoside (19), prunvuloside A (20), kaempferol 3-O-alpha-L-rhamnopyranosyl(1-->6)-beta-D-glucopranoside (21), kaempferol 3-O-beta-D-glucopyranoside (22), quercertin 3-O-alpha-L-rhamnopyranosyl(1-->6)-beta-D-glucopyranoside (23), and 2-hydroxy-3-(3',4'-dihydroxyphenly)propanoic acid (24). Compounds 1, 8-12, 17, 21, 23, and 24 were isolated from this plant source for the first time. The isolated compounds were evaluated for their cytotoxicity against A549, SK-OV-3, SK-MEL-2, and HCT15 cells in vitro using the sulforhodamin B bioassay (SRB) method. Compound 3 exhibited moderate cytotoxic activity against A549, SK-OV-3, SK-MEL-2, and HCT15 cells, with ED(50) values of 3.71, 3.65, 13.62, and 5.44 microM, respectively.