Elucidating Korean meadowsweet (Filipendula glaberrima Nakai)-derived arabinogalactan protein-induced macrophage activation and its associated mechanism of action.

This study aimed to confirm macrophage-stimulatory component from Korean meadowsweet (Filipendula glaberrima; FG) and characterize its compositional and structural properties. FG-CWH, prepared via cool-water extraction and ethanol precipitation, induced the highest secretion of NO (6.0-8.0 µM), TNF-α (8.7-9.5 ng/mL), and IL-6 (1.0-5.7 ng/mL) compared to other samples at 0.4-10 µg/mL in RAW 264.7 cells. Analytical results revealed that FG-CWH is a high-molecular-weight component with an average molecular weight of 220 kDa, constituting a polysaccharide-protein mixture. Chemical and enzymatic treatment of FG-CWH indicated its primary composition as arabinogalactan protein (AGP)-rich glycoprotein, with activity likely associated with the chemical and structural characteristics of AGP. FG-CWH treatment resulted in significant and concentration-dependent increases in iNOS (20.0-29.6 folds), TNFα (10.6-18.6 folds) and IL6 (10.9-155.6 folds) gene expression, as well as the secretion of NO (5.3-6.3 µM), TNF-α (35.4-44.3 ng/mL), and IL-6 (4.1-8.4 ng/mL) secretion, even at a reduced concentration range of 125-500 ng/mL, compared to the negative control group. Immunoblotting analysis indicated FG-CWH-induced macrophage stimulation significantly associated with the activation of MAPK (ERK, JNK, and p38) and NF-κB (p65 and IκBα). These findings can serve as valuable groundwork for developing FG-derived AGP as novel functional ingredients to enhance human immunity.

Genetic polymorphism of HLA-DRA and alcohol consumption affect hepatitis development in the Korean population.

BACKGROUND: Hepatitis is an inflammation of the liver that has several potential causes; however, the genetic association has recently begun to be studied. OBJECTIVES: Human leukocyte antigen (HLA) is an essential component of the immune response, and in this study, we conducted a correlation analysis to determine whether genetic polymorphisms of HLA and drinking habits affect hepatitis development. METHODS: Genetic polymorphisms of HLA were investigated using Korean genomic and epidemiological data. A gene association study was performed using PLINK version 1.07. Other statistical analyses and multivariate logistic regression analyses were performed using PASW Statistics version 18.0. RESULTS: Thirteen single nucleotide polymorphisms (SNPs) in HLA-DRA showed significant statistical correlations with hepatitis. In particular, rs9268645 showed the highest statistical association with hepatitis (P = 3.97 × 10-5, odds ratio [OR] = 0.72, 95% confidence interval [CI] = 0.61-0.84). In multivariate logistic regression analysis, when considering only genetic factors, the A allele of rs9268644 showed a reduced hepatitis OR of approximately 0.52-fold. However, the group carrying the minor A allele (AA + AC) with alcohol consumption had an approximately 1.58-fold OR of hepatitis compared to that of the group carrying the same allele with no alcohol consumption. This implies that the A allele of rs9268644 has a protective effect on hepatitis by genetic factors and shows sensitivity to alcohol. CONCLUSIONS: Our results showed that hepatitis is influenced by both genetic and external factors (drinking habits), which can provide new guidelines for the prevention or treatment of hepatitis.

Antioxidant and Anti-Inflammatory Activity of Filipendula glaberrima Nakai Ethanolic Extract and Its Chemical Composition.

Many countries are endeavoring to strengthen the competitiveness of their biological resources by exploring and developing wild endemic plants. This study examined the effects of Filipendula glaberrima Nakai (FG) on the antioxidant and anti-inflammatory activity using an in vitro system. The bioactive components were also examined using chromatographic techniques. The ethanol extract of Filipendula glaberrima Nakai (FGE) exerted antioxidant activities in the radical scavenging and reducing power assays and had high amounts of total polyphenolic compounds. The qRT-PCR results suggested that FGE significantly downregulated the levels cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) 2, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 in LPS-stimulated RAW 264.7 cells. The FGE treatment also decreased the production of nitric oxide, TNF-α, and IL-6 significantly in a dose-dependent manner. In addition, FGE downregulated phosphorylation of MAPK and NF-κB signaling pathway-related proteins. The chromatographic and mass spectrometry results showed that FGE contained bioactive flavonoids such as (+)-catechin, miquelianin, quercitrin, and afzelin, which may be active compounds with antioxidant and anti-inflammatory activities. This study provides fundamental data on the anti-inflammatory activity of the FG and can serve as a good starting point for developing a novel natural anti-inflammatory agent using FGE-containing bioactive flavonoids.

Genetic polymorphism of ADAM17 and decreased bilirubin levels are associated with allergic march in the Korean population.

BACKGROUND: The "allergic march" refers to changes in the frequency and intensity of allergic diseases with age. Classically, the allergic march begins with atopic dermatitis in infancy and leads to asthma and rhinitis as it continues. There are many factors that induce the allergic march; however, TNF-α may play an important role in inducing inflammation. Therefore, the therapeutic potential of TNF alpha-targeting agents is being considered for allergic march treatment. METHODS: We performed a correlation study to determine whether genetic polymorphisms of ADAM17 and clinical serum values between allergic and normal groups affect disease development by using the cohort data of the Korean genome epidemiologic research project. Gene association study was performed using PLINK version 1.07 ( http://pngu.mgh.harvard.edu/-purcell/plink ) and other statistical analysis was performed using PASW Statistics (version 18.0, SPSS Inc. Chicago, IL, USA). RESULTS: ADAM17 (also called TNF-α converting enzyme or TACE) showed a statistically significant association with the allergic march. The 13 and 8 SNPs in ADAM17 were significantly associated with asthma and allergies, respectively. Among them, on average, SNP of rs6432011 showed the greatest statistical correlation with asthma (P = 0.00041, OR = 1.95, 95% CI 1.35-2.82) and allergies (P = 0.02918, OR = 1.35, 95% CI 1.03-1.78). The effect of SNPs in ADAM17 on transcription factor binding was confirmed using RegulomeDB. The six SNPs are located in the genomic expression quantitative trait loci (eQTL) region and can affect transcription factor binding and gene expression. In clinical serum analysis, bilirubin levels were significantly decreased in the allergic group. The multivariate logistic regression analysis revealed that the low-bilirubin groups indicated a 3.22-fold increase in the prevalence of asthma compared with the high-bilirubin group. CONCLUSIONS: The ADAM17 gene and low bilirubin levels are associated with the allergic march in the Korean population, which can provide new guidelines for managing this disease progression phenomena.

Rhamnogalacturonan-I-Type Polysaccharide Purified from Broccoli Exerts Anti-Metastatic Activities Via Innate Immune Cell Activation.

To examine the anti-metastatic activities of polysaccharides in broccoli, purified polysaccharides (BCE-I, -II, and -III) were isolated by fractionation of broccoli enzyme extracts and subsequent ethanol precipitation. BCE-I mainly consisted of galactose and arabinose, whereas BCE-II mainly consisted of galacturonic acid and rhamnose, and BCE-III mainly consisted of rhamnose and galactose. Of the three fractions, stimulation of murine peritoneal macrophages by BCE-I showed the greatest enhancement of tumor necrosis factor-α, interleukin (IL)-12, and IL-6 secretion. In addition, intravenous (i.v.) administration of BCE-I enhanced the lethal activity of natural killer (NK) cells on YAC-1 tumor cells significantly and dose-dependently in an ex vivo experiment of NK cell activity. In an experimental model using lung metastasis of Colon26-M3.1 carcinoma cells, prophylactic i.v. and oral administration of BCE-I significantly and dose-dependently inhibited lung metastatic activity. Furthermore, the inhibitory activity of BCE-1 on lung metastasis partially disappeared when NK cell function was removed through treatment of rabbit anti-asialo GM1. These results indicated that BCE-I has potent antitumor metastatic activity, and that its anti-metastatic activity has relevance to the stimulation of NK and other immune cells.

Resveratrol controls Escherichia coli growth by inhibiting the AcrAB-TolC efflux pump.

Resveratrol (RSV), a phytoalexin found in grapes and other plants, is known to have antibacterial effects against Escherichia coli. In this study, we aimed to identify the target gene(s) for the antibacterial activity of RSV in E. coli. Using a DNA microarray, we found that exposure to RSV led to changes in the expression levels of iron metabolism genes, and those involved in drug response and respiration. Thus, we measured the antibacterial activity of RSV against 14 E. coli mutants with deletions in genes involved in these processes and found over fourfold higher growth inhibition in strains defective in AcrAB-TolC pump-related genes. Among the three genes encoding the AcrAB-TolC pump, tolC expression was most decreased by RSV. To determine if tolC was a direct target of RSV, we constructed both a tolC promoter-reporter gene vector and a tolC-complementation vector and transformed them into a tolC deletion mutant. RSV susceptibility and Nile red efflux tests were performed with the transformants. RSV significantly decreased tolC-promoter activity and tolC expression, thereby retarding activity of the AcrAB-TolC drug efflux complex, which may promote RSV's antibacterial activity in E. coli.

Oral administration of palatinose vs sucrose improves hyperglycemia in normal C57BL/6J mice.

Palatinose is a sucrose analog with a slower digestion rate than that of sucrose. For this reason, palatinose shows better effects on hepatic lipogenesis and cholesterol homeostasis compared with sucrose. We hypothesized that supplementation with palatinose instead of sucrose improves postprandial hyperglycemia and hyperinsulinemia in mice. Herein, we compared the digestion rates in vitro and observed physiological changes in vivo between sucrose- and palatinose-containing diets given to mice. Palatinose was hydrolyzed only by enzymes of the small intestine and was digested more slowly compared with sucrose in vitro. In mice, a diet containing palatinose resulted in significantly lower body weight gain and food efficiency rate values than those given a diet with sucrose. In this study, changes in serum biochemistry; hepatic fatty acid synthesis; cholesterol homeostasis; glucogenic, proinflammatory cytokines; and oxidative stress-related genes and proteins in the palatinose- and sucrose-fed mice were measured. Compared with the mice fed the sucrose diet, the palatinose diet resulted in lower serum glucose, insulin, and total cholesterol levels, as well as lower expression of several lipogenesis-related genes and proteins. Histological analysis of hepatic cells of palatinose-fed mice showed normal morphology. In conclusion, palatinose intake results in lower hepatic lipogenesis and better cholesterol homeostasis than the effects from sucrose.

Synergic effect in the reduction of serum uric acid level between ethanol extract of Aster glehni and vitamin B6.

In this study, synergistic hypoudpricemic activities between ethanol extract of Aster glehni (AG) and vitamin B6 were investigated in vitro and in vivo. Xanthine oxidase inhibitory activities in the different parts, leaf, stem, and flower, during spring and autumn were compared. In addition, to improve hypouricemic activity, two chemicals (AG extract and vitamins) were mixed and measured inhibitory activity of xanthine oxidase. As a result, autumn leaf AG extracts showed the most effective xanthine oxidase inhibitory activity and we named autumn leaf AG extracts as AG-D006. In synergistic study, AG-D006 with vitamin B6 showed significantly increased inhibitory activity on xanthine oxidase. AG-D006 with vitamin B6 also showed significantly reduced uric acid level in hyperuricemic rats in vivo. In conclusion, AG-D006 with vitamin B6 might be used functional foods in reducing serum uric acid level in gout.

Oxyresveratrol stimulates mucin production in an NAD+-dependent manner in human intestinal goblet cells.

The intestinal mucus layer plays an important role in the management of inflammatory bowel disease. The aim of this study was to investigate the effects of oxyresveratrol (OXY), an antioxidant, on the stimulation of mucin production in human LS 174T goblet cells and the underlying mechanism thereof. OXY increased MUC2 expression at both the mRNA and protein levels. By performing two-dimensional gel electrophoresis, we found that the expression of nicotinic acid phosphoribosyltransferase1 (NaPRT1) in OXY-treated LS 174T cells was greatly increased compared with that in negative control cells. In addition, the NAD+/NADH ratio was increased in proportion to OXY in LS 174T cells. The expression of NAD+-synthesis enzymes, NaPRT1, nicotinamide riboside kinase1 (NRK1) and nicotinamide mononucleotide adenylyltransferase1 (Nmnat1) was significantly increased at both the mRNA and protein levels in OXY-treated LS 174T cells. The inhibition of NaPRT1 and NRK1 did not decrease MUC2 expression after inhibiting by small interfering RNA (siRNA)-NaPRT1 and siRNA-NRK1, respectively; however, inhibition of Nmnat by an Nmnat inhibitor decreased MUC2 expression in a dose-dependent manner. In conclusion, OXY increases NAD+ levels, resulting in the stimulation of MUC2 expression in LS 174T cells. These findings present a novel role for NAD+ in stimulation of MUC2 expression.

Oxyresveratrol improves tight junction integrity through the PKC and MAPK signaling pathways in Caco-2 cells.

Strengthening intestinal tight junctions (TJ) provides an effective barrier from the external environment and is important for recovery from inflammatory bowel disease. Oxyresveratrol (OXY), an isomer of hydroxylated resveratrol, is isolated from many plants. The aim of this study was to investigate the effect of OXY on intestinal TJ and to elucidate the mechanism underlying the OXY-mediated increase in TJ integrity in human intestinal Caco-2 cells. OXY-treated Caco-2 cell monolayers showed decreased monolayer permeability as evaluated by paracellular transport assay. The results showed that OXY significantly increased the levels of TJ-related genes and proteins (Claudin-1, Occludin and ZO-1) compared with those of the negative control. OXY activated protein kinase C (PKC) and increased expression levels of mitogen-activated protein kinase (MAPK) genes. OXY also increased gene and protein levels of the transcription factor Cdx-2. Expression levels of TJ, PKC and Cdx-2 proteins and transepithelial electrical resistance (TEER) value decreased in OXY-treated Caco-2 cells following treatment with a pan-PKC inhibitor compared with those of the untreated control. In conclusion, OXY strengthens the integrity of the intestinal TJ barrier via activation of the PKC and MAPK pathways.

Antitumor and antimetastatic activities of rhamnogalacturonan-II-type polysaccharide isolated from mature leaves of green tea via activation of macrophages and natural killer cells.

To investigate the antitumor and antimetastatic polysaccharide from the mature leaves of green tea, GTE-II was purified using size exclusion chromatography. GTE-II consisted of 15 different sugars including rarely observed sugars such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3-deoxy-d-manno-2-octulosonic acid, and 3-deoxy-d-lyxo-2-heptulosaric acid, which were characteristics of pectic polysaccharide rhamnogalacturonan-II. Treatment of peritoneal macrophages with GTE-II not only increased interleukin (IL)-6 and IL-12 production, but also had significantly increased tumoricidal activity against Yac-1 tumor cells than those obtained from untreated mice. In an assay of natural killer (NK) cell activity, intravenous administration of GTE-II significantly stimulated NK cytotoxicity against Yac-1 tumor cells. Furthermore, the depletion of NK cells by injection of rabbit anti-asialo GM1 serum eliminated the inhibitory effect of GTE-II on B16BL6 melanoma cells. These data suggest that GTE-II inhibits tumor metastasis, and its antitumor effect is associated with activation of macrophages and NK cells.

Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells.

BACKGROUND: Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. METHODS: We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. RESULTS: OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10µg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. CONCLUSION: OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.

Bee Venom Promotes Hair Growth in Association with Inhibiting 5α-Reductase Expression.

Alopecia is an important issue that can occur in people of all ages. Recent studies show that bee venom can be used to treat certain diseases including rheumatoid arthritis, neuralgia, and multiple sclerosis. In this study, we investigated the preventive effect of bee venom on alopecia, which was measured by applying bee venom (0.001, 0.005, 0.01%) or minoxidil (2%) as a positive control to the dorsal skin of female C57BL/6 mice for 19 d. Growth factors responsible for hair growth were analyzed by quantitative real-time PCR and Western blot analysis using mice skins and human dermal papilla cells (hDPCs). Bee venom promoted hair growth and inhibited transition from the anagen to catagen phase. In both anagen phase mice and dexamethasone-induced catagen phase mice, hair growth was increased dose dependently compared with controls. Bee venom inhibited the expression of SRD5A2, which encodes a type II 5α-reductase that plays a major role in the conversion of testosterone into dihydrotestosterone. Moreover, bee venom stimulated proliferation of hDPCs and several growth factors (insulin-like growth factor 1 receptor (IGF-1R), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF)2 and 7) in bee venom-treated hDPCs dose dependently compared with the control group. In conclusion, bee venom is a potentially potent 5α-reductase inhibitor and hair growth promoter.

Resveratrol antibacterial activity against Escherichia coli is mediated by Z-ring formation inhibition via suppression of FtsZ expression.

Resveratrol exhibits a potent antimicrobial activity. However, the mechanism underlying its antibacterial activity has not been shown. In this study, the antibacterial mechanism of resveratrol was investigated. To investigate induction of the SOS response, a strain containing the lacZ+gene under the control of an SOS-inducible sulA promoter was constructed. DNA damage was measured by pulse-field gel electrophoresis (PFGE). After resveratrol treatment, the cells were observed by confocal microscopy. For the RNA silencing assay, ftsZ-specific antisense peptide nucleic acid (PNA) was used. Reactive oxygen species (ROS) production increased in Escherichia coli after resveratrol treatment; however, cell growth was not recovered by ROS quenching, indicating that, in this experiment, ROS formation and cell death following resveratrol treatment were not directly correlated. Resveratrol treatment increased DNA fragmentation in cells, while SOS response-related gene expression levels increased in a dose-dependent manner. Cell elongation was observed after resveratrol treatment. Elongation was induced by inhibiting FtsZ, an essential cell-division protein in prokaryotes, and resulted in significant inhibition of Z-ring the formation in E. coli. The expression of ftsZ mRNA was suppressed by resveratrol. Our results indicate that resveratrol inhibits bacterial cell growth by suppressing FtsZ expression and Z-ring formation.

Cherry tomato supplementation increases the area of the intestinal mucosa and the number of muscle layers in rats.

Tomatoes act as prebiotics in the gut. The effects of cherry tomatoes on gastrointestinal health have not yet been studied. Four cherry tomato supplementation diets (CTSDs) were prepared from the juice and cake of fresh and processed (heat-treated) cherry tomatoes. The contents of the gut and histological changes in the cecum and intestine were analyzed at 4weeks in rats fed CTSDs. The lactic acid bacteria level in fecal contents of rats fed CTSDs increased compared with the control. The gut length was longer in rats fed CTSDs than that in control animals. In addition, the cecal propionate level significantly increased (p<0.05), and acetate and butyrate levels decreased compared with control animals, however, regardless of the type of CTSD, the total concentration of short chain fatty acids (SCFAs) in all rats fed different CTSDs was similar with the control. The thicknesses of the mucosa and muscle of the cecum and colon increased in rats fed CTSDs compared with the control. CTSDs increased the area of the mucosa and the number of muscle layers in the intestine and cecum of rats, which strengthened the barrier function and promoted gastrointestinal health.

Effect of water-soluble fraction of cherry tomatoes on the adhesion of probiotics and Salmonella to intestinal epithelial cells.

BACKGROUND: Tomato is one of the most consumed vegetables in the world and contains many valuable nutritional components. Here we investigate the prebiotic effects of cherry tomatoes for improving gut health. RESULTS: Water-soluble dietary fiber was prepared from fresh and processed (heat treatment at 80 °C for 15 min) cherry tomato samples, each with and without Viscozyme L treatment. In the adhesion assays, all water-soluble dietary fiber samples improved adhesion of probiotics (Lactobacillus rhamnosus and Bifidobacterium bifidum) to intestinal epithelial cells (Caco-2 cells). Heat treatment in the preparation of juice from cherry tomatoes showed no significant effect on the adhesion of probiotics to Caco-2 cells. The oligofructose content of samples affected the intestinal adhesion of probiotic bacteria, with higher oligosaccharide concentrations associated with greater adhesion of probiotics and more inhibition of the adhesion of pathogens to Caco-2 cells. CONCLUSION: The present results suggest that cherry tomato can act as a prebiotic, with oligofructose potentially being one of its major prebiotic components.

The possible mechanism of rhapontigenin influencing antifungal activity on Candida albicans.

Rhapontigenin, an aglycone of rhapontin, was produced by biotransformation and we investigated its antifungal activity against Candida albicans, one of the most important opportunistic fungal pathogens. Rhapontigenin is found to have, in vitro, inhibitory activity with a minimal inhibitory concentration (MIC) value against all test isolates of 128-256 µg/ml. We detected increased reactive oxygen species (ROS) levels in yeast cultures treated with rhapontigenin at the MIC. Rhapontigenin inhibited DNA, RNA, and protein synthesis, especially RNA synthesis, and induced morphological changes and apoptosis of C. albicans. The apoptotic effect of rhapontigenin on C. albicans at subinhibitory concentrations was higher in the stationary growth phase than in the exponential phase, while the opposite results were noted with amphotericin B. The mechanism of antifungal activity of rhapontigenin may be associated with the generation of ROS that might induce apoptosis and it may also involve the inhibition of ergosterol biosynthesis.

Inhibitory effect of mulberroside A and its derivatives on melanogenesis induced by ultraviolet B irradiation.

Mulberroside A was isolated from the ethanol extract of Morus alba roots. The enzymatic hydrolysis of mulberroside A with Pectinex produced oxyresveratrol and oxyresveratrol-3-O-glucoside. We tested oxyresveratrol, oxyresveratrol-3-O-glucoside, and mulberroside A to determine whether they could inhibit ultraviolet B (UVB) irradiation-induced melanogenesis in brown guinea pig skin. Topical application of mulberroside A, oxyresveratrol, and oxyresveratrol-3-O-glucoside reduced the pigmentation in guinea pig skin. These compounds suppressed the expression of melanogenic enzymes tyrosinase, tyrosinase-related protein-1, and microphthalmia transcription factor. The anti-melanogenesis effect was highest with oxyresveratrol, intermediate with oxyresveratrol-3-O-glucoside, and lowest with mulberroside A. Mulberroside A is a glycosylated stilbene of oxyresveratrol; thus, the deglycosylation of mulberroside A resulted in enhanced inhibition of melanogenesis. Histological analysis with Fontana-Masson staining confirmed that these compounds significantly reduced the melanin content in the epidermis of UVB-irradiated guinea pig skin compared to the vehicle control. Thus, these compounds effectively reduced pigmentation and may be suitable cosmetic agents for skin whitening.