Bamboo leave extract ameliorated 12-O-tetradecanoylphorbol-13-acetate (TPA) induced ear inflammation by reducing MAP kinase levels and NF-κB activation in mice model.

Sasa coreana Nakai (SCN) is a medicinal plant commonly used against inflammation. However, the underlined mechanisms against skin inflammation is poorly understood. The present study investigated the effects of SCN leave extract on ear inflammation. To this aim, six-week-old male ICR mice was subjected to 12-O-tetradecanoyl-phorbol-13-acetate induce ear edema, which were then topically treated with the leave extract of SCN. Ear thickness, weight, and morphological changes were recorded to ensure the induction of ear edema. Further, histological analysis and protein expression for inflammatory markers were also recorded to validate the study. Topical treatment with SCN repressed TPA-induced ear edema in a dose-dependent manner. Further, SCN treatment significantly antagonized the protein expression of MAP kinase signaling pathway and reduced the effect of TPA-induced NF-κB activation, sequentially, deactivated its transcriptional targets in a dose-dependent manner. Collectively, the study suggested that SCN could be a useful therapeutic agent against skin inflammation.

Oleifolioside B-mediated autophagy promotes apoptosis in A549 human non-small cell lung cancer cells.

The biochemical mechanisms of cell death by oleifolioside B (OB), a cycloartane-type triterpene glycoside isolated from Dendropanax morbifera Leveille, were investigated in A549 human lung carcinoma cells. Our data indicated that exposure to OB led to caspase activation and typical features of apoptosis; however, apoptotic cell death was not prevented by z-VAD-fmk, a pan-caspase inhibitor, demonstrating that OB-induced apoptosis was independent of caspase activation. Subsequently, we found that OB increased autophagy, as indicated by an increase in monodansylcadaverine fluorescent dye-labeled autophagosome formation and in the levels of the autophagic form of microtubule-associated protein 1 light chain 3 and Atg3, an autophagy-specific gene, which is associated with inhibiting phospho-nuclear factor erythroid 2-related factor 2 (Nrf2) expression. However, pretreatment with bafilomycin A1, an autophagy inhibitor, attenuated OB-induced apoptosis and dephosphorylation of Nrf2. The data suggest that OB-induced autophagy functions as a death mechanism in A549 cells and OB has potential as a novel anticancer agent capable of targeting apoptotic and autophagic cell death and the Nrf2 signaling pathway.

Differential stress-response expression of two flavonol synthase genes and accumulation of flavonols in tartary buckwheat.

Flavonoids are ubiquitously present in plants and play important roles in these organisms as well as in the human diet. Flavonol synthase (FLS) is a key enzyme of the flavonoid biosynthetic pathway, acting at the diverging point into the flavonol subclass branch. We isolated and characterized a FLS isoform gene, FtFLS2, from tartary buckwheat (Fagopyrum tataricum). FtFLS2 shares 48% identity and 67% similarity with the previously reported FtFLS1, whereas both genes share 47-65% identity and 65-69% similarity with FLSs from other plant species. Using quantitative real-time PCR and high-performance liquid chromatography (HPLC), the expression of FtFLS1/2 and the production of 3 main flavonols (kaempferol, myricetin and quercetin) was detected in roots, leaves, stems, flowers and different stages of developing seeds. The relationship between the expression of the 2 FLS genes and the accumulation of the 3 basic flavonols was analyzed in 2 tartary buckwheat cultivars. FtFLS1 and FtFLS2 exhibited differential transcriptional levels between the tartary buckwheat cultivars 'Hokkai T10' and 'Hokkai T8'. Generally, higher transcript levels of FtFLS1 and FtFLS2 and a higher amount of flavonols were observed in the 'Hokkai T10' cultivar than 'Hokkai T8'. The content of flavonols showed tissue-specific accumulation between the 2 cultivars. The transcription of FtFLS1 was inhibited by the exogenous application of abscisic acid (ABA), salicylic acid (SA) and sodium chloride (NaCl), while FtFLS2 was not affected by ABA but up-regulated by SA and NaCl. These data indicate that the 2 FtFLS isoforms of buckwheat have different functions in the response of buckwheat to environmental stress.

Ganoderma lucidum ethanol extract inhibits the inflammatory response by suppressing the NF-κB and toll-like receptor pathways in lipopolysaccharide-stimulated BV2 microglial cells.

Ganoderma lucidum is a traditional Oriental medicine that has been widely used as a tonic to promote longevity and health in Korea and other Asian countries. Although a great deal of work has been carried out on the therapeutic potential of this mushroom, the pharmacological mechanisms of its anti-inflammatory actions remain unclear. In this study, we evaluated the inhibitory effects of G. lucidum ethanol extract (EGL) on the production of inflammatory mediators and cytokines in lipopolysaccharide (LPS)-stimulated murine BV2 microglia. We also investigated the effects of EGL on the LPS-induced activation of nuclear factor kappaB (NF-κB) and upregulation of toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88). Elevated levels of nitric oxide (NO), prostaglandin E(2) (PGE(2)) and pro-inflammatory cytokine production were detected in BV2 microglia following LPS stimulation. We identifed that EGL significantly inhibits the excessive production of NO, PGE(2) and pro-inflammatory cytokines, including interleukin (IL)-1ß and tumor necrosis factor-α in a concentration-dependent manner without causing cytotoxicity. In addition, EGL suppressed NF-κB translocation and transcriptional activity by blocking IκB degradation and inhibiting TLR4 and MyD88 expression in LPS-stimulated BV2 cells. Our results indicate that the inhibitory effects of EGL on LPS-stimulated inflammatory responses in BV2 microglia are associated with the suppression of the NF-κB and TLR signaling pathways. Therefore, EGL may be useful in the treatment of neurodegenerative diseases by inhibiting inflammatory mediator responses in activated microglia.

Neuroprotective effects of triterpene glycosides from glycine max against glutamate induced toxicity in primary cultured rat cortical cells.

To examine the neuroprotective effects of Glycine max, we tested its protection against the glutamate-induced toxicity in primary cortical cultured neurons. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. From such fractionation, two triterpene glycosides, 3-O-[α-l-rhamnopyranosyl(1-2)-ß-d-glucopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (1) and 3-O-[ß-d-glucopyranosyl(1-2)-ß-d-galactopyranosyl(1-2)-ß-d-glucuronopyranosyl]olean-12-en-3ß,22ß,24-triol (2) were isolated with the methanol extracts with of air-dried Glycine max. Among these compounds, compound 2 exhibited significant neuroprotective activities against glutamate-induced toxicity, exhibiting cell viability of about 50% at concentrations ranging from 0.1 µM to 10 µM. Therefore, the neuroprotective effect of Glycine max might be due to the inhibition of glutamate-induced toxicity by triterpene glycosides.

Oleifolioside A, a New Active Compound, Attenuates LPS-Stimulated iNOS and COX-2 Expression through the Downregulation of NF-κB and MAPK Activities in RAW 264.7 Macrophages.

Oleifolioside A, a new triterpenoid compound isolated from Dendropanax morbifera Leveille (D. morbifera), was shown in this study to have potent inhibitory effects on lipopolysaccharide (LPS-)stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Consistent with these findings, oleifolioside A was further shown to suppress the expression of LPS-stimulated inducible nitric oxide synthase (iNOS) and cyclooxigenase-2 (COX-2) in a dose-dependent manner at both the protein and mRNA levels and to significantly inhibit the DNA-binding activity and transcriptional activity of NF-κB in response to LPS. These results were found to be associated with the inhibition of the degradation and phosphorylation of IκB-α and subsequent translocation of the NF-κB p65 subunit to the nucleus. Inhibition of NF-κB activation by oleifolioside A was also shown to be mediated through the prevention of p38 MAPK and ERK1/2 phosphorylation. Taken together, our results suggest that oleifolioside A has the potential to be a novel anti-inflammatory agent capable of targeting both the NF-κB and MAPK signaling pathways.

Oleifolioside A mediates caspase-independent human cervical carcinoma HeLa cell apoptosis involving nuclear relocation of mitochondrial apoptogenic factors AIF and EndoG.

Apoptosis, the main type of programmed cell death, plays an essential role in a variety of biological events. Whereas "classical" apoptosis is dependent on caspase activation, caspase-independent death is increasingly recognized as an alternative pathway. To develop new anticancer agents, oleifolioside A was isolated from Dendropanax morbifera Leveille and the biochemical mechanisms of oleifolioside A-induced apoptosis in HeLa cells were investigated. Exposure to oleifolioside A resulted in caspase activation and typical features of apoptosis, although cell death was not prevented by caspase inhibition. Oleifolioside A treatment induced up-regulation of Bad, loss of mitochondrial membrane potential, nuclear relocation of mitochondrial factors, apoptosis-inducing factor (AIF), endonuclease G (EndoG), and apoptosis induction. This is the first report of anticancer activity of oleifolioside A, and nuclear translocation of AIF and EndoG in oleifolioside A-treated HeLa cells might represent an alternative death signaling pathway in the absence of caspase activity.

JNP3, a new compound, suppresses PMA-induced tumor cell invasion via NF-κB down regulation in MCF-7 breast cancer cells.

The expression of matrix metalloproteinase (MMPs)-9 is critical for cell migration and can lead to invasion and metastasis of cancer cells. In the present study, we examined the inhibitory effects of JNP3, a new compound which was isolated from traditional Chinese medicine, on cell invasion and MMP-9 activation in phorbol myristate acetate (PMA)-induced MCF-7 cells. Treatment with JNP3 significantly and selectively inhibited PMA-induced MMP-9 secretion, mRNA expression and protein levels, and these results led to reduction of cell invasion and migration in PMA-induced MCF-7 cells. The results of MMP-9 promoter assay and EMSA showed that JNP3 specifically inhibited PMA-induced MMP-9 gene expression by blocking NF-κB-dependent transcriptional activity. In addition, PMA-induced phosphorylation of ERK1/2 and JNK were suppressed by JNP3 treatment, whereas the phosphorylation of p38 MAPK was not affected by JNP3. These results suggest that JNP3 can be potential anti-cancer agents through specific inhibition of NF-κB-dependent MMP-9 gene expression.

Inhibitory effects of three oleanolic acid glycosides from Achyranthes japonica on the complement classical pathway.

The present study evaluated the anticomplement effects of isolated compounds from Achyranthes japonica in the classical pathway of the complement system. Using column chromatography, three compounds: achyranthoside C dimethyl ester, achyranthoside C butyl dimethyl ester, and achyranthoside E dimethyl ester were isolated and evaluated for in vitro anticomplement activity. Achyranthoside C dimethyl ester showed the most potent inhibitory activity against the complement system, with 50% inhibitory concentrations (IC(50)) values of 26.2 µg/mL. This is the first report of anticomplement activity of isolated compounds from A. japonica.

Anticomplement activity of organic solvent extracts from Korea local Amarantaceae spp.

The study evaluated the anticomplement activity from various solvent extracts of nine Amarantaceae plants (Achyranthes japonica (Miq.) Nakai, Amaranthus mangostanus L., Amaranthus retroflexus L., Amaranthus spinosus L., Celosia argentea var. spicata., Amaranthus lividus L., Celosia cristata L., Amaranthus viridis L., Gomphrena globosa L.) from South Korea on the classical pathway. We have evaluated various organic solvent extract from nine Amarantaceae plants with regard to its anticomplement activity on the classical pathway. Achyranthes japonica chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) value of 73.1µg/ml. This is the first report of anticomplement activity from Amarantaceae plants.

Inhibitory effects of organic solvent extracts from Korean local plants on the complement classical pathway.

The study evaluated the anticomplement effects from organic solvent extracts of five Compositae plants (Ligularia fischeri (Ledeb.) Turez, Ligularia taquetii (H.Lev. & Vaniot) Nakai, Ainsliaea acerifolia Sch.Bip, Aster scaber Thunb, Aster koraiensis Nakai, Synurus deltoides Aiton) from South Korea on the classical pathway complement system. We have evaluated organic solvent extracts from five Compositae with regard to its anticomplement activity. Chloroform extracts from L. taquetii showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 73.2 µg/mL. This is the first report of anticomplement activity from L. taquetii.

Anticomplement activity of various solvent extracts from Korea local Artemisia spp.

The study evaluated the anticomplement activity from various solvent extracts of eight Artemisia plants (Artemisia capillaris Thunb., Artemisia fukudo Makino., Artemisia japonica Thunb., Artemisia montana (Nakai) Pamp., Artemisia keiskeana Miq., Artemisia rubripes Nakai., Artemisia stolonifera (Maxim.) Kom., and Artemisia sylvatica Max.) from South Korea on the classical pathway (CP). We have evaluated various organic solvent extract from eight Artemisia plants with regard to its anticomplement activity on the CP. A. rubripes and A. montana chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC50) values of 54.3 and 64.2 µg/mL. This is the first report of anticomplement activity from Artemisia plants.

Anticomplement activity of isolated compounds from Artemisia montana.

The study evaluated the anticomplement activity from isolated compounds from Artemisia montana (Nakai) Pamp. from South Korea on the classical pathway. In a previous work, A. montana (Nakai) Pamp. chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of A. montana (Nakai) Pamp. led to the isolation of four compounds. Their structures were characterized to be ezoartemin, yamayomoginin, ezomontanin and 11,13-dihydroezomontanin by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from A. montana (Nakai) Pamp.

Inhibition effects of isolated compounds from Artemisia rubripes Nakai of the classical pathway on the complement system.

The study evaluated the anticomplement activity from isolated compounds from Artemisia rubripes Nakai from South Korea on the classical pathway. In the previous works, Artemisia rubripes chloroform extracts showed inhibitory activity against complement system. The chromatographic separation of a chloroform chloride extract of Artemisia rubripes led to the isolation of three compounds. Their structures were characterized to be scopoletin (1), 11,(13)-triene-6,12-olide (2), and 1ß,6α-dihydroxy-4(15)-eudesmene (3) by spectroscopic data. This is the first report of anticomplement activity of isolated compounds from Artemisia rubripes.

Isolated compounds from Sorghum bicolor L. inhibit the classical pathway of the complement.

The present study evaluated the anticomplement effects from isolated compounds of Sorghum bicolor in classical pathway complement system. Using column chromatograph, three compounds; Sorgoleone-362 (1), Sorgoleone-360 (2) and Sorgoleone-386 (3) were isolated and evaluated for in vitro anticomplement activity. Sorgoleone-386 showed inhibitory activity against complement system with 50% inhibitory concentrations (IC(50)) values of 148.3µg/ml. This is the first report of anticomplement activity of isolated compounds from Sorghum bicolor.

Fermented Viola mandshurica inhibits melanogenesis in B16 melanoma cells.

We assessed the effects of chloroform extract of fermented Viola mandshurica (CEFV) on melanogenesis B16 melanoma cells. CEFV treatment significantly decreased melanin content and tyrosinase activity in dose-dependent manners. To elucidate the mechanism of the inhibitory effects of CEFV on melanogenesis, we performed RT-PCR and Western blotting for melanogenesis-related genes such as tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF). CEFV strongly inhibited mRNA as well as the protein expression of tyrosinase and MITF, but had no significant effect on TRP-1 or TRP-2 expressions. It markedly decreased the phosphorylation of cAMP responsive element binding protein (CREB), and induced the duration of extracellular signal-regulated kinase (ERK) activation, leading to reduction of MITF expression and subsequently that of tyrosinase. Therefore, we suggest that CEFV induces downregulation of melanogenesis through decreased CREB phosphorylation and ERK activation.

Inhibitory effects of isolated compounds from black coloured rice bran on the complement classical pathway.

The present study evaluated the anticomplement effects of isolated compounds from black coloured rice bran in the classical pathway of the complement system. Using column chromatography, three compounds: oryzafuran, quercetin and protocatechuic acid, were isolated and evaluated for in vitro anticomplement activity. Oryzafuran showed the most potent inhibitory activity against the complement system, with 50% inhibitory concentration (IC50) values of 126.2 µg/mL. This is the first report of anticomplement activity of isolated compounds from black coloured rice bran.

Phenol glycosides with in vitro anti-Helicobacter pylori activity from Hypericum erectum Thunb.

To evaluate the anti-Helicobacter pylori activity of Hypericum erectum methanol extracts in order to provide the primary evidence for their use in clinical practice. An ethyl acetate fraction of H. erectum suspension-cell cultures inhibited the growth of H. pylori in vitro, with a MIC50 range of 38.7-63.2 µg/mL, comparable to metronidazole (MIC50 = 43.2 µg/mL). To further investigate the involved active compounds of the H. erectum extracts, four phenol glycosides were isolated for the current study: quercetin-3'-O-ß-D-galactopyranoside, quercetin-3'-O-(2''-acetyl)-ß-D-glucopyranoside, 4,6-dihydroxy-2-methoxyphenyl-1-O-ß-D-glucopyranoside and 4-hydroxy-2,6-dimethoxyphenyl-1-O-α-L-rhamnopyranosyl(1-6)-ß-D-glucopyranoside. The MIC50 values of 4,6-dihydroxy-2-methoxyphenyl-1-O-ß-D-glucopyranoside and 4-hydroxy-2,6-dimethoxyphenyl-1-O-α-L-rhamnopyranosyl(1-6)-ß-D-glucopyranoside from ATCC43504 strains were 7.3 and 27.3 µg/mL, respectively. The other two phenol glycosides did not show anti-H. pylori activity. The results of this work suggest that H. erectum has some therapeutic potential against H. pylori infection, which could be explored for patients with gastroduodenal disorders.

Protective effects of methoxyflavone derivatives from black galingale against glutamate induced neurotoxicity in primary cultured rat cortical cells.

To examine the neuroprotective effects of black galingale, its protection was tested against glutamate-induced neurotoxicity in primary cortical cultured neurons. It was found that an aqueous extract of this medicinal plant exhibited significant protection against glutamate-induced toxicity in primary cultured rat cortical cells. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. By such fractionation, bioactive methoxyflavone derivatives were isolated from the methanol extracts from the air-dried rhizomes of black galingale. 5-Hydroxy-3,7,3',4'-tetramethoxyflavone exhibited significant neuroprotective activities against glutamate-induced toxicity, exhibiting cell viability of about 60-70%, at concentrations ranging from 0.1 µm to 10 µm. Therefore, the neuroprotective effect of black galingale might be due to the inhibition of glutamate-induced toxicity by the methoxyflavone derivatives it contains.