Antiasthmatic Effects of Herbal Complex MA and Its Fermented Product MA128.

This study was conducted to determine if oral administration of the novel herbal medicine, MA, and its Lactobacillus acidophilus fermented product, MA128, have therapeutic properties for the treatment of asthma. Asthma was induced in BALB/c mice by systemic sensitization to ovalbumin (OVA) followed by intratracheal, intraperitoneal, and aerosol allergen challenges. MA and MA128 were orally administered 6 times a week for 4 weeks. At 1 day after the last ovalbumin exposure, airway hyperresponsiveness was assessed and samples of bronchoalveolar lavage fluid, lung cells, and serum were collected for further analysis. We investigated the effect of MA and MA128 on airway hyperresponsiveness, pulmonary eosinophilic infiltration, various immune cell phenotypes, Th2 cytokine production, OVA-specific IgE production, and Th1/Th2 cytokine production in this mouse model of asthma. In BALB/c mice, we found that MA and MA128 treatment suppressed eosinophil infiltration into airways and blood, allergic airway inflammation and AHR by suppressing the production of IL-5, IL-13, IL-17, Eotaxin, and OVA-specific IgE, by upregulating the production of OVA-specific Th1 cytokine (IFN-γ), and by downregulating OVA-specific Th2 cytokine (IL-4) in the culture supernatant of spleen cells. The effectiveness of MA was increased by fermentation with Lactobacillus acidophilus.

Effectiveness of the Novel Herbal Medicine, KIOM-MA, and Its Bioconversion Product, KIOM-MA128, on the Treatment of Atopic Dermatitis.

This study was conducted to determine if oral administration of the novel herbal medicine, KIOM-MA, and its Lactobacillus acidophilus-fermented product, KIOM-MA128, has therapeutic properties for the treatment of atopic dermatitis (AD). Using AD-induced BALB/c mice by Ovalbumin and aluminum hydroxide, the effectiveness of KIOM-MA and KIOM-MA128 on AD was evaluated. Oral administration of KIOM-MA and KIOM-MA128 reduced major clinical signs of AD including erythema/darkening, edema/papulation, excoriations, lichenification/prurigo, and dryness. Interestingly, KIOM-MA128 more significantly improved AD-related symptoms including decrease of IgE level in the plasma as well as reduction of scratching behavior, skin severity in the AD BALB/c model. HPLC analysis showed the significant changes in the constituent patterns between KIOM-MA and KIOM-MA128. Our results suggest that both KIOM-MA and KIOM-MA128 have potential for therapeutic reagent for the treatment of AD, and further, the efficacy is significantly enhanced by L. acidophilus fermentation via increases in its indicator molecule.

Fermentation by Lactobacillus enhances anti-inflammatory effect of Oyaksungisan on LPS-stimulated RAW 264.7 mouse macrophage cells.

BACKGROUND: Oyaksungisan (OY) has been used as a traditional drug in east-Asian countries. However, its effect on inflammation still remains unknown. In this study, to provide insight into the biological effects of OY and OY fermented by Lactobacillus, we investigated their effects on lipopolysaccharide (LPS)-mediated inflammation in the RAW 264.7 murine macrophage cells. METHODS: The investigation was focused on whether OY and fermented OYs could inhibit the production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin (PG) E2 as well as the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α, interleukin (IL)-6, nuclear factor (NF)-κB and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. RESULTS: We found that OY inhibits a little LPS-induced NO, PGE2, TNF-α and IL-6 productions as well as the expressions of iNOS and COX-2. Interestingly, the fermentation significantly increased its inhibitory effect on the expression of all pro-inflammatory mediators. Furthermore, the fermented OYs exhibited elevated inhibition on the translocation of NF-κB p65 through reduced IκBα degradation as well as the phosphorylations of extracellular signal-regulated kinase (ERK), p38 and c-Jun NH2-terminal kinase (JNK) MAPKs than untreated control or original OY. CONCLUSIONS: Finally, the fermentation by Lactobacillus potentiates the anti-inflammatory effect of OY by inhibiting NF-κB and MAPK activity in the macrophage cells.

Oploxynes A and B, polyacetylenes from the stems of Oplopanax elatus.

Two new polyacetylenes, oploxynes A (1) and B (2), and the known oplopandiol (3) and falcarindiol (4) were isolated from the stem of Oplopanax elatus. The structures of compounds 1 and 2 were determined to be 9,10-epoxyheptadeca-4,6-diyne-3,8-diol and 10-methoxyheptadeca-4,6-diyne-3,8,9-triol, respectively, on the basis of their UV, MS, and NMR data. The absolute configurations of these compounds were determined using the modified Mosher's method and acetonide formation. Oploxyne A (1), oplopandiol (3), and falcarindiol (4) inhibited the formation of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in lipopolysaccharide (LPS)-induced murine macrophage RAW 267.7 cells.

Polyacetylenes from the roots of cultivated-wild ginseng and their cytotoxicity in vitro.

Column chromatographic separation of the roots of cultivated-wild ginseng (Jangnoisam) led to the isolation of seven polyacetylenes (1-7). Their structures were determined by spectroscopic methods to be panaxynol (1), ginsenoyne-A (2), panaxydol (3), 10-methoxy heptadeca-1-ene-4, 6-dyne-3, 9-diol (4) (3R, 9R, 10R)-panaxytriol (5), panaxyne (6), and ginsenoyne-C (7). These compounds were isolated from this source for the first time. The compounds were tested for their cytotoxic activity against four human cancer cell lines (A549, SK-OV-3, SK-MEL-2, and HCT-15) in vitro using the SRB method. Panaxydol (3) and panaxyne (6) showed significant and selective cytotoxicity against SK-OV-3 with ED50 values 2.93 and 1.40 microM, respectively.

A new phenolic amide from the roots of Paris verticillata.

A new phenolic amide 8, together with the nine known phenolic compounds 1-7, 9 and 10 were isolated from the MeOH extract of the roots of Paris verticillata. The structure of the new compound 8 was determined to be 1-N-feruloylaminobutyl-4-rho-hydroxybenzamide by spectroscopic methods. The isolated compounds were tested for cytotoxicity against four human tumor cell lines using the SRB assay.

Isolation of limonoids and alkaloids from Phellodendron amurense and their multidrug resistance (MDR) reversal activity.

Three limonoids and five alkaloids were isolated from the chloroform layer of the MeOH extract of the bark of Phellodendron amurense (Rutaceae). The structures of the compounds isolated were determined to be obacunone (1), limonin (2), 12alpha-hydroxylimonin (3), gamma-fagarine (4), oxyberberine (5), canthin-6-one (6), 4-methoxy-N-methyl-2-quinolone (7) and oxypalmatine (8) based on the physicochemical and spectroscopic data. Compounds 3, 5, 7, and 8 were first isolated from the Phellodendron amurense. The isolated compounds were then tested for their cytotoxicity against five human tumor cell lines in vitro using the SRB method. Compound 5 showed significant cytotoxicity against the five tumor cell lines with ED50 values ranging from 0.30 to 3.0 microg/mL. The marginal or noncytotoxic compounds (1, 2, 3, 4, and 7) were examined for their P-gp related MDR reversal activities. Compound 1 showed significant P-gp MDR inhibition activity in MES-SA/DX5 and HCT15 cells with an ED50 value of 0.028 microg/mL and 0.0011 microg/mL, respectively.

Two new phenolic constituents of Humulus japonicus and their cytotoxicity test in vitro.

Two new phenolic constituents (4 and 6), together with four known constituents, methyl ferulate (1), eugenyl-beta-D-glucopyranoside (2), apigenin-7-O-beta-D-glucopyranoside (3), and (E)-resveratrol-3-O-beta-D-glucopyranoside (5) were isolated from the MeOH extract of the aerial part sof Humulus japonicus. The structures of the new compounds were determined by spectroscopic methods to be divarin-3-O-beta-glucopyranoside (4), and lariciresinol-9-O-beta-xylopyranoside (6). Compounds 1 and 3 exhibited moderate cytotoxicity against two human cancer cell lines (SK-OV-3 and HCT15) with ED50 values ranging from 8.84 to 8.79 microM.

Lignan and terpene constituents from the aerial parts of Saussurea pulchella.

Chromatographic separation of the MeOH extract from the aerial parts of Saussurea pulchella led to the isolation of seven terpenes (1-4, 11-13), and eight phenolics (5-10, 14-15). Their structures were determined by spectroscopic means to be (3S)-3-O-(3',4'-diangeloyl-beta-D-glucopyranosyloxy)-3,7-trimethylocta-1,6-diene (1), 7delta-methoxy- 4(14)- oppositen-1beta-ol (2) 4(15)- eudesmene-1beta, 6alpha-diol (3), 3alpha-hydroxy-5, 6-epoxy-7-megastigmen-9-one (4), (+)-syringaresinol (5), (7S, 8R, 8'R)-5,5'-dimethoxylariciresinol (6), 8alpha-hydroxypinoresinol (7), (7'R, 8'R)-2,2'- dimethoxy-4- (3-hydroxyl-propenyl)-4'-(1,2,3-trihydroxypropyl)-biphenyl ether (8), 4-allyl-2,6- dimethoxyphenyl glucoside (9), 2-methoxy-4-(2-propenyl)phenyl beta-D-glucoside (10), (-)-oplopan-4-one- 10alpha-O-beta-D-glucoside (11), linalyl-O-beta-D-glucoside (12), amarantholidoside IV (13), (+)-1-hydroxypinoresinol 1-O-beta-D-glucoside (14), and syringin (15). Compounds 1-3 and 8-13 were first isolated from the genus Saussurea. The isolated compounds were examined for cytotoxic activity against four human cancer cell lines in vitro using the sulforhodamin B bio assay method.

A new sesquiterpene glycoside from the aerial parts of Saussurea triangulata.

Column chromatographic separation of a MeOH extract of the aerial parts of Saussurea triangulata led to the isolation of a new sesquiterpene glycoside 6, together with three quinic acid derivatives, two phenolics, two sesquiterpene glycosides and two flavonoids. The new compound 6 was identified as amarantholidol A glycoside by spectroscopic and chemical methods.

Cytotoxic terpenes and lignans from the roots of Ainsliaea acerifolia.

The chromatographic separation of the MeOH extract of the roots of Ainsliaea acerifolia (Compositae) led to the isolation of six known terpenes and two known lignans. Their structures were identified by spectroscopic methods as mokko lactone (1), betulonic acid (2), betulinic acid (3), zaluzanin C (4), 1beta-hydroperoxygermacra-4(15),5,10(14)-triene (5), pluviatilol (6), (+)-syringaresinol (7), and glucozaluzanin C (8). Compounds 1 approximately 4 and 8 showed non-specific significant cytotoxicity against five human tumor cell lines with ED50 values ranging from 0.36 approximately 5.54 microg/mL.

Protoberberine alkaloids and their reversal activity of P-gp expressed multidrug resistance (MDR) from the rhizome of Coptis japonica Makino.

Six protoberberine alkaloids were isolated from the chloroform layer of the rhizome of Coptis japonica Makino (Ranunculaceae). The structures of the isolated compounds were determined to be 6-([1,3]dioxolo[4,5-g]isoquinoline-5-carbonyl)-2,3-dimethoxy-benzoic acid methyl ester (1), oxyberberine (2), 8-oxo-epiberberine (3), 8-oxocoptisine (4), berberine (5) and palmatine (6) by physicochemical and spectroscopic methods. The compound 3 (8-oxo-epiberberine) was first isolated from natural sources. The compounds were tested for cytotoxicity against five tumor cell lines in vitro by SRB method, and also tested for the MDR reversal activities. Compound 4 was of significant P-gp MDR inhibition activity with ED50 value 0.018 microg/mL in MES-SA/DX5 cell and 0.0005 microg/mL in HCT15 cell, respectively.

Cytotoxic phenolic constituents of Acer tegmentosum maxim.

The chromatographic separation of the MeOH extract from the twigs of Acer tegmentosum led to the isolation of ten phenolic compounds. The structures of these compounds were determined using spectroscopic methods as 3,7,3',4'-tetramethyl-quercetin (1), 5,3'-dihydroxy-3,7,4'-trimethoxy flavone (2), 2,6-dimethoxy-p-hydroquinone (3), (-)-catechin (4), morin-3-O-alpha-L-lyxoside (5), p-hydroxy phenylethyl-O-beta-D-glucopyranoside (6), 3,5-dimethoxy-4-hydroxy phenyl-1-O-beta-D-glucoside (7), fraxin (8), 3,5-dimethoxy-benzyl alcohol 4-O-beta-D-glucopyranoside (9) and 4-(2,3-dihydroxy propyl)-2,6-dimethoxy phenyl beta-D-glucopyranoside (10). The compounds were examined for their cytotoxic activity against five cancer cell lines. Compound 3 exhibited good cytotoxic activity against five human cancer cell lines with ED50 values ranging from 1.32 to 3.85 microM.

Phospholipids from Bombycis corpus and their neurotrophic effects.

Three phospholipids (4-6) and three aromatic amines (1-3) were obtained from the methanol extract of Bombycis corpus. Based on spectral data, their structures have been elucidated as nicotiamide (1), cytidine (2), adenine (3), 1-O-(9Z-octadecenoyl)-2-O-(8Z,11Z-octadecadienoyl) sn-glycero-3-phosphorylcholine (4), 1,2-di-O-hexadecanoyl-sn-glycero-3-phosphorylcholine (5) and 1,2-di-O-9Z-octadecenoyl-sn-glycero-3-phosphorylcholine (6). We examined the effects of compounds on synthesis of NGF in cultured astrocytes. By RT-PCR analysis, expresison of NGF mRNA in astrocytes cultured in serum-starvation increased after the addition of phospholipid (10 microM). The NGF content in the culture medium was significantly increased by compound 5, compared with the control value. These results suggest that three phospholipid compounds isolated from the methanol extract of Bombycis corpus may exert neurotrophic effects by stimulation of NGF synthesis in astrocytes.

Anti-inflammatory effect of Citrus Unshiu peel in LPS-stimulated RAW 264.7 macrophage cells.

Citrus Unshiu peel (CUP) has been traditionally used in East Asia as a drug for the treatment of vomiting and dyspepsia. However, its effects on inflammation remain unknown. In this study, we investigated the effects of CUP on the production of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The research focused on determining whether CUP could inhibit the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and the activation of nuclear factor (NF)-κB, mitogen-activated protein kinases (MAPKs), as well as the secretion of nitric oxide (NO), prostaglandin (PG) E(2), tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in LPS-stimulated RAW 264.7 cells. We found that CUP represses LPS-induced iNOS and COX-2 gene expression as well as NO, PGE(2), TNF-α and IL-6 production. Additionally, CUP inhibited the LPS-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38, and c-Jun NH(2)-terminal kinase (JNK) MAPK, and suppressed IκBα degradation and nuclear translocation of NF-κB. Collectively, our results indicate that CUP inhibits the production of various inflammatory mediators via blockade of MAPK phosphorylation pursuant to the inhibition of IκBα degradation and the nuclear translocation of NF-κB. These findings are the first to clarify the mechanism underlying the anti-inflammatory effect exerted by CUP in RAW 264.7 macrophage cells stimulated by inflammatory agents.

Fermentation improves anti-inflammatory effect of sipjeondaebotang on LPS-stimulated RAW 264.7 cells.

Sipjeondaebotang (SJ) has been used as a traditional drug in east-Asian countries. In this study, to provide insight into the biological effects of SJ and SJ fermented by Lactobacillus, we investigated their effects on lipopolysaccharide (LPS)-mediated inflammation in macrophages. The investigation was focused on whether SJ and fermented SJ could inhibit the production of pro-inflammatory mediators such as prostaglandin (PG) E(2) and nitric oxide (NO) as well as the expressions of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB in LPS-stimulated RAW 264.7 cells. We found that SJ modestly inhibited LPS-induced PGE(2), NO and TNF-α production as well as the expressions of COX-2 and iNOS. Interestingly, fermentation significantly increased its inhibitory effect on the expression of all pro-inflammatory mediators. Furthermore, fermented SJ exhibited increased inhibition of p38 MAPK and c-Jun NH(2)-terminal kinase (JNK) MAPK phosphorylation as well as NF-κB p65 translocation by reduced IκBα degradation compared with either untreated controls or unfermented SJ. High performance liquid chromatography (HPLC) analysis showed fermentation by Lactobacillus increases liquiritigenin and cinnamyl alcohol contained in SJ, which are known for their anti-inflammatory activities. Finally, SJ fermented by Lactobacillus exerted potent anti-inflammatory activity by inhibiting MAPK and NF-κB signaling in RAW 264.7 cells.

Protective effect of Ssanghwa-tang fermented by Lactobacillus fermentum against carbon tetrachloride-induced acute hepatotoxicity in rats.

Ssanghwa-tang (SHT) is a traditional herbal medicine formula that has been used for the development of physical strength, relief of pain, and the reduction of fatigue. In this study, we fermented SHT with Lactobacillus fermentum (L. fermentum), Lactobacillus gasseri (L.gasseri), or Lactobacillus casei (L.casei) to investigate the hepatoprotective effects of SHT and fermented SHT with Lactobacillus on carbon tetrachloride (CCl(4))-induced liver injury in rats. Rats were given CCl(4) (1 ml/kg, 50% CCl(4) in olive oil) intraperitoneally and either SHT or fermented SHTs (15 ml/kg) was administered 30 min before CCl(4). At 24 hr after CCl(4) injection, the levels of transaminases in the serum were markedly increased. These increases were significantly attenuated by either SHT + L. fermentum or SHT+ L.gasseri. However, SHT and SHT + L.casei showed slight suppression of the increase of transaminases. The liver histological changes were diminished by treatment with SHT + L. fermentum. Additionally, the potential hepatoprotective effect of fermented-SHTs correlated with the amount of unknown metabolite which is produced during fermentation process with L. fermentum, L.gasseri, or L.casei. Therefore, these results suggest that the hepatoprotective effect of SHT may be improved by fermentation with L. fermentum and the intestinal bacterial enzyme activities may likely play an important role in the pharmacological action of herbal medicines.

Guaiane sesquiterpene lactones and amino acid-sesquiterpene lactone conjugates from the aerial parts of Saussurea pulchella.

Two new guaiane sesquiterpene lactones ( 1 and 2) and seven new amino acid-sesquiterpene lactone conjugates ( 3- 9), together with six known sesquiterpene lactones ( 10- 15), were isolated from the methanol extract of the aerial parts of Saussurea pulchella. Their structures were determined on the basis of spectroscopic and chemical methods to be 8alpha- O-(3'-hydroxy-3'-methylbutyryl)desacylcynaropicrin ( 1), 8alpha- O-(2', 3'-dihydroxyisobutyryl)11beta,13-dihydrodesacylcynaropicrin ( 2), and pulchellamines A, B, C, D, E, F, and G ( 3- 9). The structures of the new amino acid-sesquiterpene lactone conjugates, pulchellamines A, B, C, D, E, F, and G ( 3- 9), were confirmed by synthesis. The isolated compounds were evaluated for cytotoxic activity against four human tumor cell lines. Compounds 11 and 12 exhibited cytotoxicity against skin melanoma (SK-MEL-2) and ovary malignant ascites (SK-OV-3) human tumor cell lines with ED 50 values of 1.53 and 4.07 microM, and 2.49 and 7.42 microM, respectively.