Purification of Four Flavonoid Glycosides from Lotus (Nelumbo nucifera Gaertn) plumule by Macroporous Resin Combined with HSCCC.

High-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column was successfully applied to the isolation and purification of four flavonoid glycosides from the medicinal herb Lotus plumule (LP). A polar two-phase solvent system composed of ethyl acetate-n-butanol-water (1:2:3, v/v/v) was selected by high-performance liquid chromatography (HPLC) and run on a preparative scale where the lower aqueous phase was used as the mobile phase with a head-to-tail elution mode. Quercetin-3-O-ß-D-glucopyranoside (15 mg), isorhamnetin-3-O-ß-D-glucopyranoside (13 mg), apigenin 6-C-ß-D-glucopyranosyl-8-C-α-L-arabinopyranoside (18 mg) and apigenin 6,8-di-C-ß-D-glucopyranoside (48 mg) were obtained in a one-step HSCCC separation from 240 mg of the sample. The purity of each compound was over 95% as determined by HPLC. Chemical structures of the isolated compounds were identified by electrospray ionization mass spectrometry (ESI-MS-MS) and nuclear magnetic resonance (NMR) methods. Moreover, the four compounds were isolated from LP for the first time.

Bioconversion of conjugated linoleic acid by Lactobacillus plantarum CGMCC8198 supplemented with Acer truncatum bunge seeds oil.

Conjugated linoleic acid (CLA) isomers, c9, t11-CLA and t10, c12-CLA, have been proved to exhibit excellent biomedical properties for potential use in anti-cancer applications and in reducing obesity. Acer truncatum Bunge (ATB), which is rich in unsaturated fatty acids, including oleic acid, linoleic acid, and nervonic acid, is a new resource for edible oil. In the present study, we developed a new method for producing two CLA isomers from ATB-seed oil by fermentation using Lactobacillus plantarum CGMCC8198 (LP8198), a novel probiotics strain. Polymerase chain reaction results showed that there was a conserved linoleate isomerase (LIase) gene in LP8198, and its transcription could be induced by ATB-seed oil. Analyses by gas chromatography-mass spectrometry showed that the concentration of c9, t11-CLA and t10, c12-CLA in ATB-seed oil could be increased by about 9- and 2.25-fold, respectively, after being fermented by LP8198.

Apigenin-7-O-ß-D-glucuronide inhibits LPS-induced inflammation through the inactivation of AP-1 and MAPK signaling pathways in RAW 264.7 macrophages and protects mice against endotoxin shock.

Apigenin-7-O-ß-D-glucuronide (AG), an active flavonoid derivative isolated from the agricultural residue of Juglans sigillata fruit husks, possesses multiple pharmacological activities, including anti-oxidant, anti-complement, and aldose reductase inhibitory activities. To date, no report has identified the anti-inflammatory mechanisms of AG. This study was therefore designed to characterize the molecular mechanisms of AG on lipopolysaccharide (LPS)-induced inflammatory cytokines in RAW 264.7 cells and on endotoxin-induced shock in mice. AG suppressed the release of nitric oxide (NO), prostaglandin E2 (PGE2), and tumour necrosis factor-α (TNF-α) in LPS-stimulated RAW 264.7 macrophages in a dose-dependent manner without affecting cell viability. Additionally, AG suppressed LPS-induced mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α. AG treatment decreased the translocation of c-Jun into the nucleus, and decreased activator protein-1 (AP-1)-mediated luciferase activity through the inhibition of both p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) phosphorylation. Consistent with the in vitro observations, AG protected mice from LPS-induced endotoxin shock by inhibiting proinflammatory cytokine production. Taken together, these results suggest that AG may be used as a source of anti-inflammatory agents as well as a dietary complement for health promotion.

Antibacterial active compounds from Hypericum ascyron L. induce bacterial cell death through apoptosis pathway.

Hypericum ascyron L. has been used as a traditional medicine for the treatment of wounds, swelling, headache, nausea and abscesses in China for thousands of years. However, modern pharmacological studies are still necessary to provide a scientific basis to substantiate their traditional use. In this study, the mechanism underlying the antimicrobial effect of the antibacterial activity compounds from H. ascyron L. was investigated. Bioguided fractionation of the extract from H. ascyron L. afforded antibacterial activity fraction 8. The results of cup plate analysis and MTT assay showed that the MIC and MBC of fraction 8 is 5 mg/mL. Furthermore, using Annexin V-FITC/PI, TUNEL labeling and DNA gel electrophoresis, we found that cell death with apoptosis features similar to those in eucaryon could be induced in bacteria strains after exposure to the antibacterial activity compounds from H. ascyron L. at moderate concentration. In addition, we further found fraction 8 could disrupt the cell membrane potential indicate that fraction 8 exerts pro-apoptotic effects through a membrane-mediated apoptosis pathway. Finally, quercetin and kaempferol 3-O-ß-(2″-acetyl)-galactopyranoside, were identified from fraction 8 by means of Mass spectrometry and Nuclear magnetic resonance. To our best knowledge, this study is the first to show that Kaempferol 3-O-ß-(2″-acetyl)-galactopyranoside coupled with quercetin had significant antibacterial activity via apoptosis pathway, and it is also the first report that Kaempferol 3-O-ß-(2″-acetyl)-galactopyranoside was found in clusiacea. Our data might provide a rational base for the use of H. ascyron L. in clinical, and throw light on the development of novel antibacterial drugs.

The extract of Hypericum ascyron L. induces bacterial cell death through apoptosis pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypericum ascyron L. (H. ascyron L.) has been used as a traditional medicine for the treatment of wounds, swelling, headache, nausea, stomach ache, abscesses, dysentery and chronic bronchitis. Pharmacological studies are necessary to provide a scientific basis to substantiate their traditional use. In this study, antimicrobial effect and its mechanism of the H. ascyron L. extract was investigated. MATERIALS AND METHODS: Antimicrobial activity of the H. ascyron L. extract was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT). To confirm the antimicrobial mechanism of H. ascyron L. extract on bacterial cells, Annexin V/propidium iodide (PI) double staining, TUNEL, and DNA laddering analysis were performed. RESULTS: Antibacterial effects of the H. ascyron L. extract were tested on Enterobacter cloacae, Klebsiella pneumonia, Escherichia coli, Staphylococcus aureus and Micrococcus luteus, and the results showed that the sensitive bacteria of H. ascyron L. extract mainly included E. coli, S. aureus and M. luteus, especially M. luteus. The MBC value of H. ascyron L. extract on M. luteus was equal to the MIC (20mg/mL). H. ascyron L. extract could induce cells death of M. luteus through apoptosis pathway. CONCLUSIONS: This study provides scientific support for some of the traditional uses and the pharmacological activities of H. ascyron L. Our data provide a rational base for the folkloric use of H. ascyron L.

Immunomodulatory effect of polysaccharides from submerged cultured Cordyceps gunnii.

CONTEXT: The genus Cordyceps (Clavicipitaceae) is a group of entomopathogenic fungi that is widely used as tonic food or invigorant with broad-spectrum medicinal properties in China. Cordyceps gunnii (Berk.)Berk (C. gunnii), is also well known as the Chinese rare caterpillar fungus and has similar pharmacological activities with Cordyceps sinensis (C. sinensis). Polysaccharides (PS) from various Cordyceps species have demonstrated many interesting biological activities, including antitumor, immunopotentiation, hypoglycemic, and hypocholesterolemic activities. OBJECTIVE: To investigate the effect of C. gunnii PS on the immunostimulatory antitumor function and expression of immune related cytokines in normal, immuno-suppressive, and H22-bearing mice, respectively. METHODS: C. gunnii PS were extracted with hot water at 80°C for 2 h. Normal, immuno-suppressive, and H22-bearing mice were treated with PS respectively. By detecting the value of macrophage phagocytic index, proliferation of lymphocytes, natural killer (NK) cell activity and expression of related cytokines, interleukin (IL-4), tumor necrosis factor-α (TNF-a) and interferon-γ (IFN-γ), and tumor inhibition index in H22-bearing mice additionally, the effect of PS on immunostimulatory antitumor function and its mechanism were studied. RESULTS: The total sugar content of the PS was determined to be 95% after purification. PS markedly increased the thymus and spleen indexes, the macrophage phagocytosis, the proliferation of splenic cells, and the level of IFN-γ and TNF-α. In tumor growth inhibition test, PS showed remarkable inhibition effects. CONCLUSION: PS from the C. gunnii could enhance nonspecific immunological function, humoral immunity, cellular immunity in mice, and inhibit tumor growth.

Highly efficient synthesis and antitumor activity of monosaccharide saponins mimicking components of Chinese folk medicine Cordyceps sinensis.

Ergosterol 3-O-ß-D-glucopyranoside (1a) and ergosterol 3-O-ß-D-galactopyranoside (1b) were highly efficiently synthesized and evaluated for their inhibitory activities against two tumor cell lines. The structures of these compounds were extensively confirmed by (1)H, (13)C NMR, IR, and HRMS. Compounds 1a and 1b exhibited interesting cytotoxic profiles. The antitumor activity of compound 1a was higher than that of 1b.

The chromatographic analysis of oligosaccharides and preparation of 1-kestose and nystose in yacon.

The thin-layer chromatographic analysis of the crude oligosaccharides extracted from yacon revealed the presence of glucose, fructose, sucrose, 1-kestose and nystose. The qualitative and quantitative analysis was carried out on oligosaccharides by high pressure liquid chromatography and the results showed that the contents of d-glucose, fructose, sucrose, 1-kestose, nystose and 1-fructofuranosyl nystose in oligosaccharides were 38.30%, 16.44%, 14.58%, 12.29%, 12.17%, 6.20%, respectively. The content of the fructooligosaccharides in oligosaccharides was 30.66%. The crude oligosaccharides were separated and purified by silica gel column chromatography. The two fractions obtained from crude oligosaccharides were 1-kestose and nystose, which were identified by mass spectra. The yield of 1-kestose and nystose were 10.36% and 9.73%, respectively. The purity of 1-kestose was 82.9% and of nystose was 73.6%.

Chemical analysis of a polysaccharide from Cristaria plicata (Leach).

A polysaccharide (MPS) isolated from Cristaria plicata (Leach) consisted of d-glucose. Its structural characteristics were investigated by High Performance Liquid Chromatography (HPLC), infrared analysis, gas chromatography-MS, total acid hydrolysis, methylation analysis, periodate oxidation and Smith degradation. The results indicated that the polysaccharide of C. plicata (Leach) has the weight-average molecular weight of about 2.97 × 106 Da. The structure of the polysaccharide was composed of glucose with α-(1 → 4)-linkages with short exterior chains. The fundamental information obtained from this work is beneficial to the interpretation in the relationship of the polysaccharide structure and its biological functions, and suggests that the polysaccharide from mussel may contribute to be used as a dietary supplement for health foods and therapeutics.

Antioxidant phenolic glycosides from the bark of Populus ussuriensis Kom.

Study on the EtOAc soluble fraction from the bark of Populus ussuriensis Kom. resulted in the isolation of three phenolic glycosides, including populoside (1), 7-O-ρ-coumaroylsalirepin (2) and 7-O-caffeoylsalirepin (3), among which 3 is a new compound. The structures of these compounds were elucidated on the basis of spectroscopic evidence. Phenolic glycosides 1-3 exhibited excellent antioxidant activity, evaluated in the ABTS⁺ radical scavenging assay.

Phenolic compounds in the leaves of Populus ussuriensis and their antioxidant activities.

Two new phenolic glucosides [isograndidentatin A (1) isograndidentatin B (2)], 5 known phenolic glucosides [grandidentatin (3), salireposide (4), populoside (5), populoside A (6), and salicortin (7)], and 2 known phenolic acids [P-coumaric acid (8) and caffeic acid (9)] were isolated from the leaves of Populus ussuriensis. Structure elucidation of 1 and 2 was achieved through extensive spectroscopic techniques. Compounds 1-6 and 9 showed significant antioxidant activities, which were evaluated by the DPPH radical-scavenging method (IC(50) values of 6.68, 6.61, 6.75, 6.84, 6.76, 6.79, and 5.92 microM, respectively) and the ABTS .+ radical-scavenging system (TEAC values of 1.21, 1.28, 1.26, 1.05, 1.69, 1.60, and 2.00 mM, respectively).