Objectives : In this study, the effects of ethylacetate extract of Ostericum koreanum on inflammation in RAW264.7 cells were investigated. Methods : Dried roots of Ostericum koreanum was extracted with 80% methanol for 24 h, and then fractionated with n-butanol, n-hexan and ethylacetate. RAW264.7 cells, a mouse macrophage line were incubated with different concentrations of the extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin E2 ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expressions of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX) -2 mRNA and protein were determined by RT-PCR and Western blot. Results : The methanol extract of Ostericum koreanuman and its fractions were significantly inhibited the NO and PGE2 productions in LPS-stimulated RAW264.7 cells. Among the fractions of Ostericum koreanuman the ethylacetate fraction was more strongly inhibited NO and $PGE_2$ productions compared with other fractions. The ethylacetate fraction was also suppressed LPS-induced mRNA expressions of iNOS and its protein levels in RAW264.7 cells. Conclusions : This study suggests that the ethylacetate fraction of Ostericum koreanum may have an anti-inflammatory property through suppressing inflammatory mediator productions in activated macrophages, suggesting have a therapeutic potential for the treatment of various inflammatory diseases.
The effects of micronization on in situ and in vitro nutrient disappearances of wheat, barley and corn were investigated in a series of experiments. In Experiment 1, chemical composition and in situ dry matter disappearance (DMD) of six varieties of wheat were determined. In addition, an in vitro study was completed using ground micronized and unmicronized wheat (var. Kansas). In Experiment 2, three varieties of wheat (Kansas, Sceptre and Laura) and in Experiment 3, three cereal grains (wheat, barley and corn) were either micronized for 1 min to attain internal kernel temperatures of 90-100$^{\circ}C$ or not (controls), and DM, protein and starch disappearances were estimated. In Experiment 2, an in vitro study was also completed using ground micronized and unmicronized wheat (var. Kansas). Wheat samples varied with respect to crude protein (10.0-21.2%), starch (61.6-73.9%), NDF (8.5-11.8%), volume weight (753-842 g/L) and kernel hardness (0.0-32.0). Rate (p = 0.003) and extent (p = 0.001) of in situ DMD differed among wheat varieties. Correlations between in situ kinetics, and chemical and physical properties of wheat varieties showed that protein content was negatively correlated with the rate of disappearance ($r^2$ = -0.77). Micronization of all grains markedly reduced (p = 0.001) the rate and extent of DM, and protein disappearances as compared to control samples. Micronization increased (p<0.05) the digestion of starch in wheat. However, release of ammonia into the incubation medium was markedly reduced (p<0.05), suggesting that micronization increased the resistance of protein to microbial digestion. Disappearances of DM, protein and starch differed (p = 0.001) among cereal grains with wheat>barley>corn. Micronization reduced the rate of DM disappearance (p = 0.011) and slowly degradable protein fractions (p = 0.03), however, increased (p = 0.004) slowly degradable starch fractions of all three cereals. Examination of in situ samples by scanning electron microscopy confirmed that microbial colonization focused on starch granules in micronized grains, and that the protein matrix exhibited resistance to microbial colonization. These results suggest that micronization may be used to increase the ruminal escape value of protein in cereal grains, but may lead to increased starch digestion if grains are finely ground.
To find antitumor components from the higher fungi of Korea, the mycelia of Favolus alveolarius (Fr.) Quelet were cultured in a liquid medium. The cultured mycelia were extracted with hot water twice, and a high molecular weight fraction was obtained by adding two volumes of ethanol to the extract. Two grams of Fraction A were obtained by dialyzing it. It was further separated into four fractions by gel filtration with Sepharose CL-4B, and they were designated Fractions B, C, D, and E. The results of the antitumor test showed that Fractions A, B, C, D and E had tumor inhibition ratios of 92.3, 78.5, 59.6, 77.4 and 62.2%, respectively. Anthrone test was carried out to determine the contents of total polysaccharide of the five fractions, and they had 46.3, 27.3, 65.3, 64.6, and 46.1%, respectively. The contents of the total protein of the five fractions were 29.4, 13.9, 14.3, 14.3, and 29.1%, respectively. Monosaccharide subunits of each fraction were analyzed with gas chromatography, and glucose, xylose, mannose, galactose and fucose were identified. Fraction A was examined for immunological effects. It increased the count of hemolytic plaque forming cells 12.8 times to that of the control group, and the population of macrophage in peritoneal cavity 3.2 times to that of the control group.
Kim, Eun-Jung;Yoo, Kwan-Hee;Kim, Yang-Sup;Seok, Soon-Ja;Kim, Jun-Ho
한국균학회지
/
제45권3호
/
pp.175-187
/
2017
Laetiporus sulphrueus var. miniatus is widely distributed worldwide, and has commonly been used as a medicinal mushroom. In the present study, we investigated the effects of water extract and solvent fractions from the Laetiporus miniatus as possible antioxidant, anti-thrombin and anti-invasive agents against phorbol 12-myristate 13-acetate (PMA)- or thrombin-induced matrix metalloproteinase-2 (MMP-2) and MMP-9 activities. Samples were fractionated into n-hexane, $CHCl_3$, ethyl acetate, n-butanol, and water fractions, and individually analysed. The water fraction had the highest extraction yield at 34.90% (w/w), while the n-butanol fraction demonstrated the highest anti-oxidative activity at 81.44%. In the thrombin inhibitory activity test, the water fraction exhibited the highest activity at 94.64%. Even at the concentration of $40{\mu}g/mL$, evaluation of anti-proliferating activity in YD-10B cells did not reveal any cytotoxic effects. Although MMP-9 expression in YD-10B cells increased after the addition of PMA and thrombin, MMP-2 did not. Additionally, MMP-2/-9 levels in PMA-treated YD-10B cells (i.e., both mRNA expression and protein activation) were highly inhibited in the hexane and chloroform fractions. Compared with MMP-2 levels, MMP-9 mRNA expression and proteolytic activity were inhibited to a greater extent by the hexane and chloroform fractions in thrombin-treated YD-10B cells. Taken together, these results support that thrombin induces tumor invasion through MMP-2/9 and suggest that the L. miniatus may act as an effective functional food, conferring anti-oxidative, anti-thrombotic and anti-cancer activities.
In order to determine the extent of change in nitrogen fractions and in vitro ruminal degradability of forage protein during ensilage and the influence on nitrogen utilization by sheep, orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) were ensiled in separate 120 L silos for 5, 21 and 56 days. With respect to nitrogen fractions, fraction 1 (buffer solution soluble nitrogen), fraction 2 (buffer solution insoluble nitrogen-neutral detergent insoluble nitrogen), fraction 3 (neutral detergent insoluble nitrogen-acid detergent insoluble nitrogen), and fraction 4 (acid detergent insoluble nitrogen) were determined. Fractions 1 and 2 accounted for more than 80% of total nitrogen in orchardgrass and 90% of that in alfalfa. The proportion of fraction 1 in orchardgrass increased from 33.0% at day 0 to 52.0% after day 56 of ensiling. In the case of alfalfa silage it was 41.7% and 62.9%, respectively. Seventy percent of this increase occurred within the first 5 days of ensiling. A similar change of in vitro ruminal degradability of total nitrogen was also observed in both forages. Nitrogen retention in sheep tended to decrease as the length of ensiling increased, with a significantly positive correlation between urinary nitrogen and fraction 1, and in vitro ruminal degradability of total nitrogen.
Fluorescence properties and carbohydrate content were investigated using ultrafiltrated size fractions of dissolved organic matters (DOM) originated from different sources. The materials included a treated sewage, an algal organic matter, and a soil leachate, all of which are major constituents of dissolved organic matter in a typical urban river. Four different size fractions were separated from the three sources of each DOM. The size distribution demonstrated that a higher molecular weight fraction was more present in soil leachate compared to two other source DOMs. A higher content of carbohydrates was observed in the following order - algal DOM > treated sewage > soil leachate. A wide range of specific UV absorbance was observed from size fractions of a single source DOM, indicating that aromatic carbon structures are heterogeneously distributed within one source of DOM. The structural heterogeneity was the most pronounced for the soil leachate. The fluorescence index ($F_{450}/F_{500}$) of the treated sewage was similar to that (2.0) typically obtained from autochthonous DOM, suggesting that the treated sewage exhibited autochthonous organic matter-like properties. No protein-like fluorescence intensities were observed for all of the soil leachate size fractions whereas they were observed with two other source DOMs. Based upon the fluorescence peak ratios from fluorescence excitation-emission matrix (EEM), two discrimination indices could be suggested to distinguish three different source DOMs. It is expected that the suggested discrimination indices will be useful to predict the sources of DOM in a typical urban river affected by treated sewage.
This study was carried out to develop antitumor agents based on effects of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos on human oral epitheloid carcinoma cells. Human oral epitheloid carcinoma cells were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with A, B, C, D, and E fractions for 48hrs under the same condition. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay were performed. The light microscopic findings were observed by inverted microscope. In MTT assay, fraction B was shown significant antitumor activity (P<0.001), fraction E was shown significant antitumor activities (P<0.05), but the other fractions were not shown. In NR assay, fraction B was shown significant antitumor activity (P<0.001). In SRB assay, fractions B was shown significant antitumor activities (P<0.01). fractions A and D were shown significant antitumor activities (P<0.05). but the other fractions were not shown. In light microscopy. the fraction B of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos showed the highest antitumor activity. These finding suggested that fraction B possessed the most antitumorous agent.
Physicochemical properties, intestinal microbial growth, and inhibitory effects of alcohol-insoluble polysaccharide (AIP) extracted from cucumber peel were investigated. AIP was composed of 14.54% crude protein, 1.04% crude lipid, 13.74 % crude ash, 9.1% soluble dietary fiber, and 41.2% insoluble dietary fiber. AIP showed low bulk density (0.18 g/mL) and water-holding capacity (6.39 g/g), and high oil-holding capacity (3.96 g/g). Pectic substance fractions [water-soluble pectic substance (WSP), ethylenediaminetetraacetic acid-soluble pectic substance (ESP), and alkali-soluble pectic substances (ASP)] and hemicellulose fractions [1 M KOH-soluble hemicellulose (KHP1) and 4 M KOH-soluble hemicellulose (KHP4)] were obtained from sequential chemical fractionation of AIP. WSP showed higher total sugar contents than total uronic acid contents, whereas opposite results were observed in ESP and ASP. Molecular weight distributions of three pectic substance fractions were in order of ASP>ESP>WSP. Ion exchange chromatogram pattern of WSP was different from those of ESP and ASP. Major component of WSP was fraction eluted by 0.05 M ammonium acetate buffer, whereas that of ESP and ASP was fraction eluted by 0.2 M NaOH. WSP and ASP showed growth-promoting activities against Lactobacillus brevis, Bifidobacterium bifidum, and B. longum, whereas B. bifidum and B. longum for ESP. KHP1 and KHP4 fractions had significant growth-promoting activities against B. bifidum.
We investigated the nutritional characteristics and antioxidant effects of sea mustard Undaria pinnatifida fractions from Gultongmeori in Taejongdae, Youngdo, Busan. Based on dry weight, the moisture, crude protein, crude lipid, crude ash, and crude fiber contents were 34.98%, 11.55%, 0.43%, 17.82%, and 3.45%, respectively. To evaluate the antioxidant effect, we used radical scavenging (DPPH and ABTS), reactive oxygen species (ROS) production measurement, and DNA oxidation assays. Total flavonoid and phenol contents were higher in the n-hexane fraction than in other fractions. The n-hexane fraction was more effective at scavenging free radicals than other fractions as assessed using DPPH and ABTS assays (P<0.05). The ROS production assay showed that all sea mustard fractions decreased H2O2 induced cellular ROS production compared to that seen in the control (P<0.05); however, the n-hexane fraction reduced cellular ROS production to a greater extent than the other fractions. Furthermore, the n-hexane fraction from Gultongmeori significantly inhibited genomic DNA oxidation. These results indicate that the antioxidant effect of sea mustard is associated with its high flavonoid and phenol content. This study proposes that processed food products supplemented with sea mustard can be developed as functional foods to promote health in the local population.
Objectives : Agrimoniae Herba is a herbal medicine widely distributed in Asia and contains flavonoids including catechin, quercitrin, rutin, hyperoside, and quercetin. This study aimed to investigate the anti-oxidant activity and skin barrier function of different solvent fractions (Hexane; methylene chloride, MC; ethyl acetate, EA; n-butanol, Bu; Water) obtained from Agrimoniae Herba. Methods : Anti-oxidant activity of different solvent fractions obtained from Agrimoniae Herba was investigated through total polyphenol contents, total flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity measurements. Then, filament aggregating protein (Filaggrin), Type I collagen, ceramide synthase (CERS) 3, and CERS4 were analyzed to evaluate the skin barrier strengthening effect of different solvent fractions obtained from Agrimoniae Herba on UVB-stimulated HaCaT cells. Results : As a result of measuring total polyphenol contents, total flavonoid contents, DPPH free radical scavenging activity, and ABTS radical scavenging activity, antioxidant activity was found to be excellent in the order of EA > Bu > MC > Hexane > Water. As a result of measuring mRNA gene expression of Type I collagen, Filaggrin, CERS3, and CERS4 after UVB-stimulated was applied to HaCaT cells treated with different solvent fractions obtained from Agrimoniae Herba, it was found to increase significantly in the Bu-treated group. Conclusion : Our findings show that the Bu sample obtained from Agrimoniae Herba has excellent anti-oxidant ability, which increases Type I collagen, Filaggrin, and ceramide synthetase in UVB-stimulated HaCaT cells to control the skin barrier improvement effect.
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