MOESM4 of A novel coumarin, (+)-3′-angeloxyloxy-4′-keto-3′,4′-dihydroseselin, isolated from Bupleurum malconense (Chaihu) inhibited NF-κB activity

Additional file 4. 1H NMR spectrum of Pd-Ib

Immunosuppressive Decalin Derivatives from Red Yeast Rice

Five new decalin derivatives (<b>1</b>–<b>5</b>), together with two known compounds (<b>6</b> and <b>7</b>), were isolated from the ethyl acetate extract of red yeast rice. Their structures were elucidated by means of NMR and mass spectroscopic analyses. Monascusic lactone A (<b>1</b>) is the first reported naturally occurring decalin derivative possessing a spiro lactone at the C-1 position. The immunosuppressive effects of all these isolates (<b>1</b>–<b>7</b>) on human T cell proliferation were investigated, and all, especially monascusic acids B (<b>2</b>), C (<b>3</b>), D (<b>4</b>), and A (<b>6</b>) and heptaketide (<b>7</b>), suppressed human T cell proliferation in a dose-dependent manner from 10 to 100 μM. This is the first report on the immunosuppressive activity of decalin derivatives

Cytotoxic Dehydromonacolins from Red Yeast Rice

Two new dehydromonacolins (<b>1</b> and <b>3</b>), together with nine known monacolins (<b>4</b>–<b>12</b>), were isolated from red yeast rice. Compounds <b>4</b>–<b>6</b> were isolated from a natural resource for the first time. Their structures were elucidated by means of NMR and mass spectroscopic analyses. The structure of dehydromonacolin N (<b>1</b>) was further confirmed by its semisynthesis from monacolin K (lovastatin) (<b>11</b>). Dehydromonacolin J (<b>2</b>), an intermediate in the semisynthesis of <b>1</b>, was obtained as a new dehydromonacolin. The structure of dehydromonacolin L (<b>3</b>) was also confirmed by an elimination reaction of monacolin L (<b>12</b>). Compound <b>1</b>, possessing a C2 side chain, is unprecedented in the natural monacolin family and exhibited moderate cytotoxic activity against Hep G2, Caco-2, and MCF-7 cancer cell lines. Dehydromonacolin K (<b>8</b>) demonstrated the most potent cytotoxicity to all three of these cell lines. The structure–activity relationship of natural and synthesized monacolins was discussed. This is the first report on the cytotoxic effects of dehydromonacolins

Structure Elucidation and Immunomodulatory Activity of A Beta Glucan from the Fruiting Bodies of <i>Ganoderma sinense</i>

<div><p>A polysaccharide named GSP-2 with a molecular size of 32 kDa was isolated from the fruiting bodies of <i>Ganoderma sinense</i>. Its structure was well elucidated, by a combined utilization of chemical and spectroscopic techniques, to be a β-glucan with a backbone of (1→4)– and (1→6)–Glc<i>p</i>, bearing <i>terminal-</i> and (1→3)–Glc<i>p</i> side-chains at <i>O</i>-3 position of (1→6)–Glc<i>p</i>. Immunological assay exhibited that GSP-2 significantly induced the proliferation of BALB/c mice splenocytes with target on only B cells, and enhanced the production of several cytokines in human peripheral blood mononuclear cells and derived dendritic cells. Besides, the fluorescent labeled GSP-2 was phagocytosed by the RAW 264.7 cells and induced the nitric oxide secretion from the cells.</p></div

Stimulating effect of GSP-2 on the proliferation of the mouse splenocytic B and T cells.

<p>Freshly fractionated splenocyte, splenic B (solid bars) and T (open bars) cells were stimulated with, or without (<i>Medium</i>), dextran or GSP (30 µg/ml). LPS (2 µg/ml) and ConA (2 µg/ml) were included as controls. In a parallel experiment, mouse splenocytes were stimulated with, or without (<i>Medium</i>), GSP (30 µg/ml), dextran (30 µg/ml) or LPS (10 µg/ml) in triplicate wells in the presence, or absence, of PMB. ³H-TdR was added to the cultures for the last 8 hrs of incubation and then ³H-TdR incorporation (CPM) of each well counted. A: Parallel experiment to exclude the influence of the endotoxin contamination; B: Stimulating effect of GSP-2 to the mouse splenocytic B and T cells. All results are presented as mean ± SEM, *, P<0.05; **, P<0.01; ***, P<0.001 for difference from culture without treatment. (n = 9, repeated 3 times).</p

Nitric oxide production and phagocytosis of GSP-2-treated RAW 264.7 cells.

<p>A: The levels of NO production of RAW264.7 cells were assessed after incubation with GSP-2 or LPS for 24 h using Griess assay. All data are expressed as mean ± SEM of three individual experiments (n = 12). Significant difference: *, P<0.05; **, P<0.01; ***, P<0.001 for difference from culture without treatment. B: The viable RAW 264.7 cell populations were gated (left) and the FITC-positive population was shown in fluorescent-GSP-2-labeled cells (lower right histogram). C: RAW 264.7 cells were incubated with 10 µg GSP-2 (fluro-labeled) for 24 h. The cells were imaged at the 1^st and 24^th hour (lower histograms). After 24 h incubation, the cells were collected, washed and resuspended in PBS and the fluorescence of the samples was detected by flow cytometry. (Upper histograms: viable RAW 264.7 cells without fluorescent labeled GSP-2, lower histograms, 1^st hour: viable RAW 264.7 cells with fluorescent labeled GSP-2, 24^th hour).</p

GC–MS test result for the methylated sugar moieties of the polysaccharide GSP-2 ^a^,^b.

a<p>The result were tested on DB-5 GC-MS column.</p>b<p>All the sugar residues were primarily identified by their MS spectrum and further confirmed by their relative retention time to 2,3,4,6-Me4-Glc.</p

Cytokine productions of GSP-2-treated PBMCs.

<p>Culture supernatants were collected 24-2 and the cytokines concentrations were specifically determined by ELISA. All data are expressed as mean ± SEM of three individual experiments (n = 12). Differences between the treated and untreated control groups were compared using one-way ANOVA. * P<0.05, ** P<0.01, *** P<0.001 for difference from culture without treatment.</p

HPLC profile, methylation analysis result and 1D NMR spectrums of GSP-2.

<p>A: HPLC profile of GSP-2. Samples (2 mg/mL, 10 µL) were analyzed on an Agilent 1100 system equipped with an ELSD detector and TSK GMPW_XL gel filtration columns (7.8×300 mm×2), with 20 mM CH₃COONH₄ as mobile phase at 0.6 mL/min and column temperature at 40°C. Commercially available T-series dextrans (MW 2000, 670, 410, 270, 150, 80, 50, 12, 5 and 1 kD). B: Methylation analysis result of GSP-2. GC-MS tests for methylation analysis were measured with a DB-5 column (30 m×0.25 mm×0.25 µm), and at temperatures programmed from 170–225 °C at 2 °C/min and then hold on 10 min, in the figure, a: <i>T</i>-Glc<i>p</i>, b: <i>1,3</i>-linked Glc<i>p</i>, c: <i>1,4</i>-linked Glc<i>p</i>, d: <i>1,6</i>-linked Glc<i>p</i>, e: <i>1,3,6</i>-linked Glc<i>p</i>. C-E: 1D NMR (¹H, ¹³C, DEPT) spectra of GSP-2 in D₂O with TMS as external standard, obtained on a Bruker AM 500 spectrometer with a dual probe in the FT mode at room temperature.</p

2D NMR spectrums and proposed structure of the polysaccharides from the fruiting bodies of <i>G. sinense</i>.

<p>(A: HSQC spectrum, B: HMBC spectrum, C: NOESY spectrum, D proposed strcuture).</p