Dammarane triterpenes and phytosterols from Dysoxylum tpongense Pierre and their anti-inflammatory activity against liver X receptors and NF-κB activation.

Dysoxylum tpongense Pierre (local name 'Huynh Dan Bap') belonging to family Meliaceae, is a tree (3-10 m height), distributed in the mountainous areas (ca. 1000 m a.s.l.) in North Vietnam. From the dichloromethane fraction of the methanol extract of the leaves and stems of this plant, six dammarane triterpenes, one furanoid diterpene together with three sterols were isolated. Evaluation of biological activities of isolated compounds showed that cabraleahydroxylactone (5), cabraleahydroxylactone 3-acetate (6), and stigmast-4-en-3-one (10) possessed an anti-inflammatory effect against Liver X receptor (LXR) activation in HepG2 cell line model with IC50 values of 20.29 ± 3.69, 24.32 ± 2.99, and 7.09 ± 0.97 (µM), respectively. While three other triterpenoid compounds aglinin C 3- acetate (1), aglinin C (2), and 24-epi-cabraleadiol (4) presented the most significant inhibitory effect against TNF-α induced NF-κB activation in HepG2 cell line in a dose-dependent manner with IC50 values of 12.45 ± 2.37, 23.32 ± 3.25, and 13.95 ± 1.57 µM, respectively. As stigmast-4-en-3-one (10), with structure closely similar to cholesterol, acted selectively on LXRs but not on NF-kB activation pathway, this suggests that stigmast-4-en-3-one (10) can be potentially applied as an agonist on LXR signaling pathway. Pathways LXRs-NF-κB-iNOS expression have a close relationship and play a crucial role in proceeding metabolic abnormalities like atherosclerosis, obesity, inflammation, etc. Thus, the findings showed that dammarane-type triterpenoids from D. tpongense are worthy of further investigation for potential LXR agonists and potent anti-atherogenic agents against atherosclerotic lesion progression.

Nuclear Factor Kappa B Activation and Peroxisome Proliferator-activated Receptor Transactivational Effects of Chemical Components of the Roots of Polygonum multiflorum.

BACKGROUND: Polygonum multiflorum is well-known as "Heshouwu" in traditional Chinese herbal medicine. In Northeast Asia, it is often used as a tonic to prevent premature aging of the kidney and liver, tendons, and bones and strengthening of the lower back and knees. OBJECTIVE: To research the anti-inflammatory activities of components from P. multiflorum. MATERIALS AND METHODS: The compounds were isolated by a combination of silica gel and YMC R-18 column chromatography, and their structures were identified by analysis of spectroscopic data (1D, 2D-nuclear magnetic resonance, and mass spectrometry). The anti-inflammatory activities of the isolated compounds 1-15 were evaluated by luciferase reporter gene assays. RESULTS: Fifteen compounds (1-15) were isolated from the roots of P. multiflorum. Compounds 1-5 and 14-15 significantly inhibited tumor necrosis factor-α-induced nuclear factor kappa B-luciferase activity, with IC50 values of 24.16-37.56 µM. Compounds 1-5 also greatly enhanced peroxisome proliferator-activated receptors transcriptional activity with EC50 values of 18.26-31.45 µM. CONCLUSION: The anthraquinone derivatives were the active components from the roots of P. multiflorum as an inhibitor on inflammation-related factors in human hepatoma cells. Therefore, we suggest that the roots of P. multiflorum can be used to treat natural inflammatory diseases. SUMMARY: This study presented that fifteen compounds (1-15) isolated from the roots of Polygonum multiflrum exert signifiant anti inflmmatory effects by inhibiting TNF α induced NF κB activation and PPARs transcription. Abbreviation used: NF κB: Nuclear factor kappa B, PPARs: Peroxisome proliferator activated receptors, PPREs: Peroxisome proliferator response elements, TNF α: Tumor necrosis factor α, ESI-MS: Electrospray ionization mass spectrometry, HepG2: Human hepatoma cells.

A new phenolic derivative with soluble epoxide hydrolase and nuclear factor-kappaB inhibitory activity from the aqueous extract of Acacia catechu.

One novel phenolic compound, (4S,5R)-4-(3,4-dihydroxyphenyl)-5-(3-oxobutyl)dihydrofuran-2(3H)-one (1), as well as 12 known compounds (2-13) was obtained from the aqueous extract of Acacia catechu and their chemical structures were determined by spectroscopic analysis. Compounds 8 and 9 exhibited significant soluble epoxide hydrolase (sEH) inhibitory activities with IC50 values of 26.6 ± 0.5 and 24.4 ± 5.6 µM, respectively. Compounds 7-10 showed significant inhibitory effects on TNFα-induced nuclear factor kappa B (NF-κB) transcriptional activity in a dose-dependent manner, with IC50 values ranging from 11.15 to 19.45 µM.

Interleukin-18 enhances breast cancer cell migration via down-regulation of claudin-12 and induction of the p38 MAPK pathway.

Interleukin-18 (IL-18) was recently reported to have a pro-tumor effect in various cancers. Increased IL-18 levels in the serum of cancer patients correlated with malignancy, and IL-18 acts a crucial factor for cell migration in gastric cancer and melanoma. Claudins, which are the most important tight junction proteins, are also linked with cancer progression and metastasis. However, the relationship between claudins and IL-18 is not well-understood. Here, we show that the migratory ability of MCF-7 cells was reduced when endogenous IL-18 expression was inhibited with IL-18 siRNA. Moreover, exogenous IL-18 enhanced breast cancer cell migration and suppressed the expression of the tight junction proteins claudin-1, claudin-3, claudin-4, and claudin-12 in MCF-7 cells. Knockdown of claudin-3, claudin-4, and claudin-12, but not claudin-1, increased breast cancer migration with maximal effects observed in claudin-12 siRNA-transfected cells. To investigate whether the mitogen-activated protein kinase (MAPK) signaling pathway is involved in IL-18-induced cell migration and claudin-12 expression, cells were pretreated with SB203580 (an inhibitor of p38 MAPK) or PD98059 (an inhibitor of ERK1/2) prior to the addition of IL-18. Although pretreatment of MCF-7 cells with SB203580 blocked both the enhanced cell migration and the decreased claudin-12 expression, PD98059 only blocked cell migration and did not affect claudin-12 expression. In addition, exogenous IL-18 induced rapid phosphorylation of p38 MAPK. These results suggest that IL-18 is an important factor inducing breast cancer cell migration through down-regulation of claudin-12 and activation of the p38 MAPK pathway.

Peroxisome proliferator-activated receptor transactivational effects in HepG2 cells of cembranoids from the soft coral Lobophytum crassum Von Marenzeller.

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that regulate the expression of multiple genes involved in metabolic, anti-inflammatory, and developmental processes. This study evaluated the PPARs transactivational effects of thirteen cembranoid diterpenoids 1-13 from the soft coral Lobophytum crassum, using PPAR-responsive elements-luciferase reporter and GAL4-PPAR chimera assays. All isolated compounds activated the transcription of PPARs in a dose-dependent manner, with EC50 values ranging from 2.07 ± 1.73 to 130.20 ± 1.85 µM. Moreover, compounds 6-9 affected the transactivation of all three PPAR types, PPARα, γ, ß(δ), in a dose-dependent manner, with EC50 values in a ranging from 11.92 ± 1.23 to 122.50 ± 2.12 µM. These results provide a scientific rationale for further studies on the soft coral L. crassum and its diterpenoid constituents to develop medicinal products against inflammatory and metabolic diseases.

Sterol fatty acid esters from the mushroom Hericium erinaceum and their PPAR transactivational effects.

Six new (erinarols A-F, 1-6) and five known (7-11) ergostane-type sterol fatty acid esters were isolated from the methanol extract of the dried fruiting bodies of Hericium erinaceum. Their chemical structures were elucidated using chemical and physical methods as well as through comparison of NMR and mass spectral data with those reported previously. This is the first comprehensive investigation on ergostane-type sterol fatty acid esters from H. erinaceum. The isolated compounds were evaluated for their PPAR transactivational effects using a luciferase reporter system. Compounds 1 and 2 significantly activated the transcriptional activity of PPARs in a dose-dependent manner, with EC50 values of 8.2 and 6.4 µM, respectively. Moreover, compounds 1 and 2 also activated PPARα and PPARγ transcriptional activity, with stimulation from 1.3- to 3.9-fold at 20 µM concentrations.

Antitumor effects of dammarane-type saponins from steamed Notoginseng.

Six dammarane-type saponins were extracted from steamed Panax notoginseng. Their chemical structures were identified spectroscopically as ginsenosides Rh1 (1), Rg1 (2), 20 (S)-Rg3 (3), 20 (R)-Rg3 (4), Rb3 (5), and Rb1 (6). Compounds (0.1-10 µM) were tested for inhibition of tumor necrosis factor-α (TNF)-induced nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) luciferase reporter activity using a human kidney 293T cell-based assay. Ginsenoside Rb3 (5) showed the most significant activity with an IC50 of 8.2 µM. This compound also inhibited the induction of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) messenger Ribonucleic acid (mRNA) in a dose-dependent manner after HepG2 cells had been treated with TNF-α (10 ng/mL).

Phenolic components from the stem of Acanthopanax koreanum and their inhibitory effects on NF-kappa B.

Two novel phenolic glucosides (1-2), as well as seven known compounds (3-9), were isolated from the stem of Acanthopanax koreanum; their chemical structures were determined by chemical and spectroscopic methods and subsequently compared with previously reported data. Their inhibition of nuclear factor kappa B (NF-κB) was measured in human embryonic kidney (293T) cells by using an NF-κB luciferase assay.

NF-κB inhibitory activity of polyoxygenated steroids from the Vietnamese soft coral Sarcophyton pauciplicatum.

Chromatographic purification of the methanolic extract from the soft coral Sarcophyton pauciplicatum led to the isolation of three polyhydroxylated steroids 1-3, including a new compound, sarcopanol A (1). Their structures were elucidated by spectroscopic analysis and by comparison of the spectroscopic data with those of similar compounds previously reported in literature. The anti-inflammatory effects of isolated compounds were evaluated using nuclear factor kappa B (NF-κB) luciferase and reverse transcription polymerase chain reaction (RT-PCR). The effect of isolated compounds on cell growth was evaluated by MTS assays. Compounds 1 and 2 significantly inhibited TNFα/INFγ-induced NF-κB transcriptional activity in human keratinocyte (HaCaT) cells in a dose-dependent manner, with EC50 values of 8.27±3.28 and 26.07±5.59 µM, respectively. Furthermore, the transcriptional inhibition of these compounds was confirmed by a decrease in cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and intercellular adhesion molecule-1 (ICAM-1) gene expression levels in HaCaT cells.

The Nuclear Orphan Receptor NR4A1 is Involved in the Apoptotic Pathway Induced by LPS and Simvastatin in RAW 264.7 Macrophages.

Macrophage death plays a role in several physiological and inflammatory pathologies such as sepsis and arthritis. In our previous work, we showed that simvastatin triggers cell death in LPS-activated RAW 264.7 mouse macrophage cells through both caspase-dependent and independent apoptotic pathways. Here, we show that the nuclear orphan receptor NR4A1 is involved in a caspase-independent apoptotic process induced by LPS and simvastatin. Simvastatin-induced NR4A1 expression in RAW 264.7 macrophages and ectopic expression of a dominant-negative mutant form of NR4A1 effectively suppressed both DNA fragmentation and the disruption of mitochondrial membrane potential (MMP) during LPS- and simvastatin-induced apoptosis. Furthermore, apoptosis was accompanied by Bcl-2-associated X protein (Bax) translocation to the mitochondria. Our findings suggest that NR4A1 expression and mitochondrial translocation of Bax are related to simvastatin-induced apoptosis in LPS-activated RAW 264.7 macrophages.

Inhibition of TNF-α-Mediated NF-κB Transcriptional Activity by Dammarane-Type Ginsenosides from Steamed Flower Buds of Panax ginseng in HepG2 and SK-Hep1 Cells.

Panax ginseng is a medicinal herb that is used worldwide. Its medicinal effects are primarily attributable to ginsenosides located in the root, leaf, seed, and flower. The flower buds of Panax ginseng (FBPG) are rich in various bioactive ginsenosides, which exert immunomodulatory and anti-inflammatory activities. The aim of the present study was to assess the effect of 18 ginsenosides isolated from steamed FBPG on the transcriptional activity of NF-κB and the expression of tumor necrosis factor-α (TNF-α)-stimulated target genes in liver-derived cell lines. Noticeably, the ginsenosides Rk3 and Rs4 exerted the strongest activity, inhibiting NF-κB in a dose-dependent manner. SF and Rg6 also showed moderately inhibitory effects. Furthermore, these four compounds inhibited the TNF-α-induced expression of IL8, CXCL1, iNOS, and ICAM1 genes. Consequently, ginsenosides purified from steamed FBPG have therapeutic potential in TNF-α-mediated diseases such as chronic hepatic inflammation.

NF-κB inhibitory activities of glycosides and alkaloids from Zanthoxylum schinifolium stems.

Zanthoxylum schinifolium is an aromatic shrub, the pericarp and leaves of which are widely used in culinary applications in East Asian countries. In the present study, one new neolignan glycoside, zanthoxyloside A (1) together with 16 known glycosides (2-12) and alkaloids (13-17), were isolated from methanol extract of the stems of Z. schinifolium. The absolute configuration of one known monoterpenoid glycoside (2) was determined. The structures of the isolated compounds were established by one dimensional (1D), 2D NMR and mass spectrometry. The nuclear factor-κB (NF-κB) inhibitory activities of the isolated compounds stimulated with tumor necrosis factor alpha (TNFα) were measured using a luciferase reporter system. Compounds 1, 5, 16, and 17 exhibited significant inhibition of NF-κB activation in a dose-dependent manner. Furthermore, compounds 1, 5, 16, and 17 inhibited TNFα-induced expression of inducible nitric oxide synthase (iNOS) and intercellular adhesion molecule-1 (ICAM-1) mRNA and dose-dependent inhibition of iNOS promoter activity.

Cembranoid diterpenes from the soft coral Lobophytum crassum and their anti-inflammatory activities.

Nine cembranoid diterpenes 1-9, including four new compounds, crassumols D-G (1-4), were isolated from the methanol extract of the Vietnamese soft coral Lobophytum crassum. Spectroscopic methods were used to elucidate the structures of these compounds. Compound 5 exhibited a potent inhibitory effect on tumor necrosis factor-alpha (TNFα)-induced nuclear factor-kappa B (NF-κB) transcriptional activation in HepG2 cells and significantly inhibited the mRNA expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in a dose-dependent manner.

New anti-inflammatory cembranoid diterpenoids from the Vietnamese soft coral Lobophytum crassum.

Four new cembranoid diterpenes lobocrasols A-D (1-4), were isolated from the methanol extract of the soft coral Lobophytum crassum. Their structures were elucidated by spectroscopic analysis and by comparison of the spectroscopic data with those of similar compounds previously reported in literature. The anti-inflammatory effects of isolated compounds were evaluated using NF-κB luciferase and reverse transcription polymerase chain reaction (RT-PCR). Compounds 1 and 2 significantly inhibited TNFα-induced NF-κB transcriptional activity in HepG2 cells in a dose-dependent manner, with IC50 values of 6.30±0.42 and 6.63±0.11µM, respectively. Furthermore, the transcriptional inhibition of these compounds was confirmed by a decrease in cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) gene expression levels in HepG2 cells.

Inhibition of NF-κB transcriptional activation in HepG2 cells by diterpenoids from the soft coral Sinularia maxima.

Anti-inflammatory transcriptional effects of nineteen compounds (1-19) from the soft coral Sinularia maxima were evaluated using NF-κB luciferase and reverse transcriptase polymerase chain reaction. Compounds 1, 2, 4, 8, 15, 17, and 18 significantly inhibited TNFα-induced NF-κB transcriptional activity in HepG2 cells in a dose-dependent manner, with IC50 values ranging from 15.81 ± 2.29 to 29.10 ± 1.54 µM. Furthermore, the transcriptional inhibitory function of these compounds was confirmed by a decrease in intercellular adhesion molecule-1 and inducible nitric oxide synthase gene expression levels in HepG2 cells. These results provide a scientific rationale for the use of the soft coral S. maxima warrant further studies to develop new agents for the prevention and treatment of inflammatory.

NF-κB inhibitory activity of sucrose fatty acid esters and related constituents from Astragalus membranaceus.

Twelve compounds, including six sucrose fatty acid esters (1-6), four galactosyl acylglycerols (7-10), and two sphingolipids (11 and 12), were isolated from the roots of Astragalus membranaceus . Their structures were identified on the basis of spectroscopic analysis. Among the isolated sucrose fatty acid esters, 6'-O-linoleyl sucrose (1) was identified as a new compound, and 6'-O-palmitoyl sucrose (2) and 6-O-palmitoyl sucrose (3) were isolated from nature for the first time. This is the first report on sucrose fatty acid ester components from A. membranaceus. The nuclear factor-κB (NF-κB) inhibitory activity of isolated compounds was measured in HepG2 cells stimulated with TNF-α using a luciferase reporter system. Among them, compounds 1-6 exhibited significant inhibition of NF-κB activation in a dose-dependent manner, with IC50 values ranging from 4.4 to 24.7 µM. Compounds 1-6 also exhibited inhibition of TNF-α-induced expression of iNOS and ICAM-1 mRNA and dose-dependent inhibition of iNOS promoter activity, with IC50 values ranging from 3.3 to 5.0 µM. These data demonstrate the potential of sucrose fatty acid esters from A. membranaceus to prevent and treat inflammatory diseases.

Sterols isolated from seeds of Panax ginseng and their antiinflammatory activities.

BACKGROUND: Panax ginseng C. A. Meyer, a perennial herb from the Araliaceae family, is a commonly used medicinal plant. Many studies have been conducted on the biologically active constituents of whole parts of P. ginseng (i.e., roots, leaves, flower buds, and fruits). However, the seeds of P. ginseng have not been intensively investigated. A new sterol glucoside,3-O-b-d-glucopyranosyl-5,22,24-stigmastatrienol (1), and a known sterol, 5,22-stigmastadienol (2), were isolated from seeds of P. ginseng and were evaluated for their inhibitory activities on tumor necrosis factor (TNF)α-induced nuclear factor (NF)-κB and inducible nitric oxide synthase (iNOS) transcription in transfected HepG2 cells. The present work deals with the isolation, identification, and antiinflammatory activities of the two compounds. MATERIALS AND METHODS: The compounds were isolated by a combination of silica gel and YMC R-18 column chromatography, and their structures were identified by analysis of spectroscopic data (1D, 2D-NMR, and MS). The antiinflammatory activities of the isolated compounds 1 and 2 were evaluated by luciferase reporter gene assays. RESULTS: Two sterols have been isolated from the seeds of P. ginseng. Compound 1 is a previously unreported glucosidyl sterol. Compounds 1 and 2 both inhibited NFκB-luciferase activity, with IC50 values of 8.1 and 4.8΅M, respectively. They also inhibited iNOS-luciferase activity in TNFα-induced HepG2 cells, with IC50 values of 2.2 and 2.9΅M, respectively. CONCLUSION: The two isolatedsterols have inhibitory effects on inflammation-related factors in HepG2 cells, as determined by luciferase reporter gene assays. Thus, seeds of P. ginseng are worthy of consideration for the development and research of antiinflammatory agents.

Oleanane-triterpenoids from Panax stipuleanatus inhibit NF-κB.

In continuation of our research to find biological components from Panax stipuleanatus, four oleanane-type triterpenes (12 to 15) were isolated successively. Fifteen oleanane-type saponins (1 to 15) were evaluated for nuclear factor (NF)-κB activity using a luciferase reporter gene assay in HepG2 cells. Compounds 6 to 11 inhibited NF-κB, with IC50 values between 3.1 to 18.9 µM. The effects on inducible nitric oxide synthase and cyclooxygenase-2 by compounds 8, 10, and 11 were also examined using reverse transcription-polymerase chain reaction. Three compounds (8, 10, and 11) inhibited NF-κB activity by reducing the concentration of inflammatory factors in HepG2 cells.

A new lupane-type triterpene from the seeds of Panax ginseng with its inhibition of NF-κB.

A new lupane-triterpene, 3ß-cis-feruloyloxy-16ß-hydroxylup-20(29)-ene (1) were isolated from the ethyl acetate extract of Panax ginseng seeds along with the known compound, 3ß-transferuloyloxy-16ß-hydroxylup-20(29)-ene (2). Compound 2 was isolated from this plant for the first time. Their chemical structures were determined by mass spectroscopy and one-dimensional and two-dimensional magnetic resonance spectra. The bioactive effects of these compounds on TNF-α-induced NF-κB transcription were evaluated in transfected HepG2 cells. Effects on the expression of NF-κB target genes were also examined using a reverse transcription-polymerase chain reaction. Both compounds 1 and 2 were inhibited NF-κB activity in HepG2 cells by decreasing the cellular concentrations of inflammatory factors iNOS and COX2.

Anti-inflammatory and PPAR transactivational effects of secondary metabolites from the roots of Asarum sieboldii.

Phytochemical study on the roots of Asarum sieboldii resulted in the isolation of one new compound, (1R,2S,5R,6R)-5'-O-methylpluviatilol (1) and 12 known compounds (2-13). Their structures were determined by extensive spectroscopic methods, including 1D and 2D NMR, and MS spectra. The absolute configuration of compound 1 was established using CD spectrum. Compounds 4, 5, and 12/13 significantly inhibited TNFα-induced NF-κB transcriptional activity in HepG2 cells in a dose-dependent manner, with IC(50) values ranging from 6.4 to 9.4 µM. Furthermore, the transcriptional inhibitory function of these compounds was confirmed based on decreases in COX-2 and iNOS gene expression in HepG2 cells. Compounds 1-3, 6,7, 10, and 11 significantly activated the transcriptional activity of PPARs in a dose-dependent manner, with EC(50) values ranging from 1.7 to 20.9 µM. Compounds 7, 10, and 11 exhibited significant dose-dependent PPARα transactivational activity, with EC(50) values of 19.5, 15.7, and 4.0 µM, respectively. Compounds 1, 6, 7, 10, and 11 activated PPARγ transcriptional activity, with EC(50) values ranging from 3.6 to 22.6 µM, whereas compounds 10 and 11 significantly increased PPARß(δ) transactivational activity, with EC(50) values of 22.6 and 4.9 µM, respectively. These results provide a scientific support for the use of the roots of A. sieboldii and warrant further studies to develop new agents for the prevention and treatment of the inflammatory and metabolic diseases.