Isoprenoid flavonoids isolated from Sophora davidii and their activities induces apoptosis and autophagy in HT29 cells.

Four previously undescribed isoprenoid flavonoids (2-5) were isolated from Sophora davidii, along with five known analogues. The structures of the compounds were established through comprehensive analysis of spectroscopic data, including HRESIMS, 1D and 2D NMR, and absolute configurations determined by theoretical calculations, including ECD and NMR calculation. The cytotoxic effects of the isolated compounds on human HT29 colon cancer cells were evaluated using the MTT assay, compound 1 exhibited cytotoxicity against human HT29 colon cancer cells with an IC50 value of 8.39 ± 0.09 µM. Studies conducted with compound 1 in HT29 cells demonstrated that it may induce apoptosis and autophagy in HT29 by promoting the phosphorylation of P38 MAPK and inhibiting the phosphorylation of Erk MAPK.

Breaking barriers: How modified citrus pectin inhibits galectin-8.

Inhibition of galectin-3-mediated interactions by modified citrus pectin (MCP) could affect several rate-limiting steps in cancer metastasis, but the ability of MCP to antagonize galectin-8 function remains unknown. We hypothesized that MCP could bind to galectin-8 in addition to galectin-3. In this study, a combination of gradual ethanol precipitation and DEAE-Sepharose Fast Flow chromatography was used to isolate several fractions from MCP. The ability of these fractions to antagonize galectin-8 function was studied as well as the primary structure and initial structure-function relationship of the major active component MCP-30-3. The results showed that MCP-30-3 (168 kDa) was composed of Gal (13.8%), GalA (63.1%), GlcA (13.0%), and Glc (10.1%). MCP-30-3 could specifically bind to galectin-8, with an MIC value of 0.04 mg mL-1. After MCP-30-3 was hydrolyzed by ß-galactosidase or pectinase, its binding activity was significantly reduced. These results provide new insights into the interaction between MCP structure and galectin function, as well as the potential utility in the development of functional foods.

Diterpenoids from Torreya grandis and their cytotoxic activities.

Eight previously undescribed diterpenoids, along with eleven previously reported analogues, were obtained from the supercritical CO2 extracts of Torreya grandis aril. The structures of these compounds were elucidated based on HRESIMS, NMR, ECD, and single-crystal X-ray diffraction data. In the MTT assay, compound 18 exhibited significant inhibitory effects on two human colon cancer cell lines, HT-29 and HCT 116 cells, with IC50 values of 7.37 µM and 6.55 µM, respectively. It was found that compound 18 induced apoptosis and significantly inhibited the migration of HCT 116 colon cancer cells in a concentration-dependent manner.

Structural elucidation of lactone derivatives from Dendrobium pendulum.

Six lactone derivatives, including four α-pyrones derivatives (1-4), two α-furanone derivatives (5 and 6), were isolated from the Dendrobium pendulum. Structural elucidation of these undescribed lactone derivatives were accomplished on the basis of detailed nuclear magnetic resonance analysis, and the absolute configurations of compounds 1-4 were confirmed by electronic circular dichroism (ECD) techniques. The cytotoxic effects of isolated compounds on human breast cancer cell MDA-MB-231 were evaluated by the MTT assay.

Complete 1 H and 13 C assignments of two new sesquiterpenoids from Dendrobium aduncum.

Two new sesquiterpenoids, dendroaduoid A (1) and dendroaduol (2), together with four known sesquiterpenoids were isolated from the stems of Dendrobium aduncum. Their structures were identified by HR-ESI-MS and NMR experiments, and the complete assignments of 1 H and 13 C NMR data for two new sesquiterpenoids were obtained by the aid of HSQC, HMBC, 1 H-1 H COSY, NOESY, and ECD techniques. The cytotoxic effects of the isolated compounds on four tumor cell lines (HCT-116, HepG2, A549, and SW1990) were evaluated using MTT assay. Otherwise, the inhibitory activity of these six sesquiterpenoids on glycosidase was also evaluated.

Dihydrophenanthrenes and phenanthrenes from Dendrobium terminale.

Two previously undescribed dihydrophenanthrene derivatives (1 and 2) were isolated along with twelve known analogues from the whole plant of Dendrobium terminale. The structures of the new compounds were elucidated on the basis of detailed spectroscopic analysis. The NMR data of known phenanthrene derivatives (7 and 9) were revised by 2D NMR. The isolated compounds were evaluated for cytotoxicity against three kinds of tumor cell lines (sw1990, HCT-116, and HepG2). Especially compounds 11 and 14 showed stronger antitumor effects, and the structure-activity relationship of these compounds was discussed.

Dendrobine-type alkaloids from Dendrobium nobile.

Six dendrobine-type alkaloids were isolated from the tubes of Dendrobium nobile by silica gel, Sephadex LH-20 gel column chromatography, and preparative HPLC. Compound 1 is a new alkaloid containing a pair of amide tautomers, whereas compound 2 is a new dendrobine-type alkaloid. By using spectroscopic techniques including 1 D and 2 D NMR, the structures of compounds 1‒6 were identified as N-methoxylcarbonyldendrobine (1), dendronboic acid (2), dendrobine (3), 6-hydroxyldendrobine (4), dendrobine N-oxide (5), and denrine (6). The cytotoxic effects of the isolated compounds on two human tumour cell lines (HCT-116 and SW1990) were evaluated using MTT assay.

Purification, chemical analysis and inhibitory effects on galectin-3 of enzymatic pH-modified citrus pectin.

Modified citrus pectin (MCP), a commercially available dietary supplement prepared from citrus pectin, contains several different polysaccharide domains, but its primary chemical structure and the binding epitopes that antagonize galectin-3 function remain unclear. In this study, five fractions were isolated from MCP after endo-polygalacturonase degradation (EMCP) and a combination of DEAE-cellulose and Sepharose CL-6B or Sephadex G-75 chromatography. Their primary structures, abilities to inhibit galectin-3-mediated hemagglutination, and antiproliferation activities on MCF-7 and A549 cell lines were studied. Results showed that EMCP-3p, one of the five fractions, was composed of Glc (89.8%), Gal (3.8%), Ara (3.1%), GalA (1.1%), Man (0.9%), and Rha (1.3%) with an average molecular weight of 88.4 KDa, which had the most substantial degree of galectin-3 inhibition with an MIC of 31.25 µg/mL, and it exhibited remarkable cytotoxicity against MCF-7 (36.7%) and A549 (57.4%) cell lines. These results provide new insight into the structure-function relationships of EMCP-derived polysaccharides.

Cardioprotective effects of Amentoflavone by suppression of apoptosis and inflammation on an in vitro and vivo model of myocardial ischemia-reperfusion injury.

Inflammation modulation is currently considered a promising therapeutic strategy to counteract the burden of cardiovascular disease. Amentoflavone (AME) is a natural biflavone with two apigenin molecules that, possess promising anti-inflammatory, anti-oxidative, and anti-cancer properties. In the present study, we aimed to investigate the effects of AME on myocardial ischemia-reperfusion injury in vivo and in vitro, and to elucidate the underlying mechanism. Our results showed that AME significantly reduced the levels of LDH, CK-MB, IL-6, IL-1ß, and TNF-α after hypoxia (H) 12 h/reoxygenation (R) 4 h treatment, and significantly increased the cell survival rate of H9c2 cardiomyocytes induced by H/R and inhibited their apoptosis rate. AME (25, 50, 100 mg·kg-1·d-1, i.g.) or a positive control drug diltiazem (DIZ) (16 mg·kg-1·d-1, i.g.) was used as pretreatment for 7 days; the myocardial ischemia-reperfusion(I/R) model was established. TTC staining results showed that the infarct volume was significantly reduced after AME and DIZ treatment. Oral administration of AME dose-dependently ameliorated I/R injury-induced increase in pro-inflammatory factors (IL-6, IL-1ß, and TNF-α) and levels of LDH and CK-MB. Results of TUNEL and HE staining showed that the I/R model had more induced apoptosis, but could be effectively reduced by pretreatment with AME. After surgery, the heart of the rat was examined via western blotting to detect inflammation-related proteins. Compared with the sham group, the p-AKT in the I/R group was significantly reduced and the content of p-NF-κBp65 was significantly increased. However, these changes could be reversed by AME treatment. DIZ treatment exerted similar beneficial effects in I/R rats as the high dose of AME did. This study highlights the excellent therapeutic potential of AME for managing myocardial ischemia-reperfusion injury.

A new triterpenoids from Nothotsuga longibracteata.

A new lanostane triterpenoid (1) and two known (2, 3) analogues were isolated from Nothotsuga longibracteata. Their chemical structures were identified by spectral data including HR-ESI-MS, 1 D, and 2 D NMR. These lanostane triterpenoids showed no cytotoxic activities against three human tumour cell lines (A172, SHSY5Y, and Hela), but exhibited the activity of promoting the gastrointestinal motility of zebrafish treated with Nile red.

Simultaneous determination of six bioactive components of total flavonoids of Scorzonera austriaca in rat tissues by LC-MS/MS: application to a tissue distribution study

ABSTRACT A liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of six bioactive constituents including vitexin, orientin, isoorientin, 2"-O-β-D-xylopyranosyl isoorientin, 2"-O-β-D-xylopyranosyl isovitexin, and 6-C-L-α-arabipyranosyl vitexin in rats' various tissues using isoquercitrin as the internal standard. Biological samples were pretreated by protein precipitation with acetonitrile. Chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and 0.1% aqueous formic acid. All analytes and internal standard were quantitated through electrospray ionization in negative ion selected reaction monitoring mode. The mass transitions were as follows: m/z 431 → 311 for vitexin, m/z 447 → 327 for orientin or isoorientin, m/z 579 → 459 for 2"-O-β-D-xylopyranosyl isoorientin, m/z 563 → 293 for 2"-O-β-D-xylopyranosyl isovitexin, m/z 563 → 353 for 6-C-L-α-arabipyranosyl vitexin, and m/z 463 → 300 for the internal standard, respectively. The lower limits of quantification for the six analytes in different tissue homogenates were 0.8-2.2 ng/ml. The validated assay was successfully applied to a tissue distribution study of the six components in rats after intravenous administration of total flavonoids of Scorzonera austriaca Willd; Asteraceae. The results of the tissue distribution study showed that the high concentrations of six components were mainly in the kidney.

A Cascade-Targeting Nanocapsule for Enhanced Photothermal Tumor Therapy with Aid of Autophagy Inhibition.

Autophagy is a homeostatic lysosome-dependent metabolic process to eliminate damaged or dysfunctional cellular organelles, which is closely associated with tumor progression. Indocyanine green (ICG) can convert NIR light energy to localized heat for cancer cell and tissue ablation. However, the effect of autophagy modulation on ICG-mediated photothermal therapy remains unknown. In this study, it is found that primaquine (PQ) suppresses autophagy flux at a late stage through the impeding fusion of the autophagosome with the lysosome to form an autophagolysosome, leading to cell apoptosis or necrosis. This autophagosome-lysosome fusion inhibitory effect and the autophagosome accumulation are more evident in the photothermal therapy combined with autophagy inhibition. Motivated by this notable effect, a cascade-targeting nanocapsule (HCP) is constructed using an organic solvent-free strategy to coencapsulate PQ and ICG. By targeting the cluster designation 44 molecule and sequentially enhancing the cell-penetrating peptide-mediated endocytosis, the codelivery of PQ/ICG by HCP achieves selective recognition and reinforces the internalization by MCF-7 cells to exert a synergistic therapeutic effect on MCF-7 cells both in vitro and in vivo. The HCP system for the photothermal and autophagy inhibition combination therapy represents a novel strategy for the treatment of breast cancer.

New Triterpenoid Saponins from the Herb Hylomecon japonica.

Background: Hylomecon japonica, a plant of the Papaveraceae family which is well-known for the alkaloids they produce, is a perennial plant widely distributed in the northeast, central and east regions of China. Although a variety of chemical constituents, including alkaloids, flavonoids, and megastigmoids, have been isolated from H. japonica, the investigation of saponins in H. japonica has not been reported until now. Methods: Various separation techniques, including polyporous resin column chromatography, silica gel column chromatography and hemi-preparative HPLC were applied to the isolation of triterpenoid saponins, and chemical methods such as acid hydrolysis and spectroscopic methods including HRESIMS and NMR were applied to their structure elucidation, and the XTT reduction method was used to assay cytotoxicity. Results: Two new triterpenoid saponins, named hylomeconoside A (1) and B (2) which were identified as 3-O-ß-d-galactopyranosyl-(1→2)-ß-d-glucuronopyranosyl-gypsogenin-28-O-ß-d-xylopyranosyl-(1→3)-ß-d-xylopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→2)-ß-d-quinovopyranoside (1) and 3-O-ß-d-galactopyranosyl-(1→2)-ß-d-glucuronopyranosyl-gypsogenin-28-O-ß-d-xylopyranosyl-(1→3)-ß-d-xylopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→2)-α-l-arabinopyranoside (2), and two known triterpenoid saponins identified as dubioside C (3) and lucyoside P (4) on the basis of spectroscopic and chemical evidence, were isolated from H. japonica. Compound 1 exhibited moderate cytotoxicity on MGC-803 and HL-60 cells, with IC50 values of 43.8 and 32.4 µg·mL-1, respectively. Conclusions: Compounds 1 and 2 are new saponins, and 1 is considered to be one of the antitumor principles in this plant. This is the first time that triterpenoid saponins have been isolated from plants of the Papaveraceae family.

Scarless Wound Closure by a Mussel-Inspired Poly(amidoamine) Tissue Adhesive with Tunable Degradability.

Burn, trauma, and various medical conditions including bacterial infection, diabetes complication, and surgery could lead to an acute cutaneous wound and scar formation. Application of tissue glues instead of sutures could minimize the additional trauma and scar formation. Despite the countless efforts devoted to the development of high-strength tissue glues, little attention has been paid to their influence on the scar formation. Here, we report the development of a new tissue glue with excellent biocompatibility and tunable degradability for scarless wound closure. A series of catechol-containing poly(amidoamine) (CPAA) polymers were synthesized via the one-step Michael addition of dopamine and bisacrylamide. The tertiary amino group in the polymer backbone was used to introduce a zwitterionic sulfobetaine group by one-step ring-opening polymerization. The introduction of the zwitterionic sulfobetaine group could easily tune the hydrophilicity and the degradability of CPAA without influencing the density of the catechol group in the polymer. Lap-shear tests on the porcine skin demonstrated a high adhesion strength of 7 kPa at 1 h, rising to 24 kPa by 12 h. Addition of silica nanoparticles could further enhance the adhesion strength by 50%. In vivo studies further confirmed that the CPAA tissue glue could effectively accelerate the healing process of incisional wounds on the back of Sprague Dawley rats compared with suture and reduce the scar formation.

The effective mechanism of the polysaccharides from Panax ginseng on chronic fatigue syndrome.

Ginseng acidic polysaccharide WGPA isolated from the root of Panax ginseng C. A. Meyer was fractionated into WGPA-A and WGPA-N by anion-exchange chromatography. The antifatigue activity of ginseng acidic polysaccharide WGPA has been reported in our previous research. This present study was designed to identify its active component and elucidate the mechanism for preventing chronic fatigue syndrome (CFS). WGPA, WGPA-A and WGPA-N were orally administered to mice once daily for 15 days. The effects of these compounds on physiological biomarkers of oxidative stress and on the morphology of the mitochondria in striated skeletal muscle were assessed. The results of forced swimming test-induced indicated that WGPA and WGPA-A could lengthen the swimming time, while WGPA-N could not. In addition, malondialdehyde and lactate dehydrogenase levels in serum were enhanced; while those of superoxide dismutase and glutathione peroxidase were lowered. Interestingly, the structural degeneration of mitochondria were all ameliorated. These findings suggested that WGPA-A is the active component of WGPA, it might have potential therapeutic effects for CFS and the oxidative stress might be involved in the pathogenesis. Our results also provided essential data for a better understanding of the antifatigue effects of P. ginseng extracts.

Neuroprotective effects of ginseng pectin through the activation of ERK/MAPK and Akt survival signaling pathways.

In this study, we investigated the neuroprotective activities of ginseng pectin (GP) against hydrogen peroxide (H2O2)-induced neuronal toxicity in different neuronal cells. GP selectively attenuated H2O2-induced damage up to 26% in primary cortical neuron cells and human glioblastoma U87 cells. Following H2O2 exposure, DAPI staining and neuron-specific ß-tubulin antibody probing indicated that GP maintained cell integrity and decreased nuclei condensation. Data from western blot analysis revealed that pre-treatment with GP increased the phosphorylation of both the extracellular signal-regulated kinases 1 and 2 (ERK1/2) and Akt in cortical neuron cells. However, the phosphorylation of ERK1/2 was increased, but that of Akt was decreased in U87 cells. These results suggest that the protective effects of GP against H2O2-induced apoptosis may be due to the activation of the phosphorylation of ERK1/2 and Akt; however, the mechanisms involved differ depending on the cell line. This neuroprotective property indicates that GP could serve as a potential therapeutic agent for neurodegenerative diseases.

Preparation of a glucan from the roots of Rubus crataegifolius Bge. and its immunological activity.

A water-soluble glucan (RCP-1) was prepared from the roots of Rubus crataegifolius Bge. by extraction with hot-water, deproteination by Sevag reagent, alpha-amylase treatment and ultrafiltration. RCP-1 consisted of only glucose, and its molecular weight was determined to be approximately 7KD by high performance gel permeation chromatography (HPGPC). Fourier transform infra-red spectroscopy (FT-IR), nuclear magnetic resonance spectroscopy (NMR), methylation and periodate oxidation analyses indicated that RCP-1 was an alpha-d-glucan. Its main chains were composed of (1-->4)- and (1-->6)-linked alpha-glucopyranosyls, and side chains were single alpha-glucopyranosyl residues attached to the O-6 of glucosyls in the main chains. RCP-1 could increase both cytotoxic activity against B16 melanoma cells and the production of nitric oxide (NO) of macrophages in vitro. Furthermore, in vivo bioassay tests indicated that RCP-1 could remarkably enhance T and B lymphocyte proliferations, augment the phagocytosis of macrophages and increase the tumour necrosis factor-alpha (TNF-alpha) levels in serum.