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Crocetin is a main bioactive component with a carotenoid skeleton in Gardenia jasminoides, a typical traditional Chinese medicine with a long history in Southeast Asia. Crocetin is being commonly consumed as spices, dyes, and food colorants. Recent pharmacological studies had implied that crocetin may possess potent anti-inflammatory properties; however, the underlying molecular mechanism is not fully elucidated. In the present study, the regulatory effect of crocetin on redox balance was systematically investigated in lipopolysaccharide- (LPS-) stimulated RAW264.7 cells. The results showed that crocetin dose-dependently inhibited LPS-induced nitric oxide production and inducible nitric oxide synthase (iNOS) expression in RAW264.7 cells. Molecular data revealed that crocetin exerted its anti-inflammatory property by inhibiting the MEK1/JNK/NF-κB/iNOS pathway and activating the Nrf2/HO-1 pathway. The shRNA-knockdown (KD) of MEK1 and ERK1 confirmed that the activation of MEK1 and inhibition of JNK mediated the anti-inflammatory effect of crocetin. Moreover, the pull-down assay and computational molecule docking showed that crocetin could directly bind to MEK1 and JNK1/2. It is noticed that both KD and knockout (KO) of HO-1 gene blocked this action. More detailed data have shown that HO-1-KO blocked the inhibition of p-IκB-α by crocetin. These data indicated that crocetin exerted its anti-inflammatory property via modulating the crosstalk between the MEK1/JNK/NF-κB/iNOS pathway and the Nrf2/HO-1 pathway, highlighting HO-1 as a major player. Therefore, the present study reveals that crocetin can act as a potential candidate for redox-balancing modulation in charge of its anti-inflammatory and chemopreventive effect, which strengthens its potency in the subsequent clinic application in the near future.

Many studies have shown that flavonoids have various physiological activities such as anti-inflammatory, antibacterial, anti-cancer, and hepatoprotective activities. Meanwhile, its hydrophilic nature has led to low absorption into the body and the limitations of its bioactivity. Many acylated flavonoids and their bioactivities, including those obtained as natural compounds and those synthesized biochemically or chemically, have been reported to improve their bioavailability. This review focuses on the recent developments of the cancer chemopreventive activity of acylated flavonoids. Novel acylated flavonoids have been identified from nature since 2015, and many of them have shown high cytoprotective and anti-inflammatory effects by acylation of glycosyl residues. A large number of acylated flavonoids have been synthesized by enzyme-catalyzed and chemical methods, and their bioactivity has been evaluated. The introduction of acyl groups in the sugar residues showed higher antioxidant activity with increased lipophilicity and improved anti-inflammatory and cytoprotective effects and the ability of apoptosis induction in cancer cells. In contrast, introducing acyl groups into the hydroxy groups of aglycones resulted in less antioxidant activity but improved cancer proliferation inhibition and anti-inflammatory effects. Acylation of flavonoids tends to increase their bioactivity over the parent compound. The increased bioactivity by acylation can be attributed to increased lipophilicity and improved membrane permeability. Acylated flavonoids are one of a group of promising compounds that have cancer chemopreventive effects. However, the study of ADME (Absorption, Distribution, Metabolism, Excretion) will be essential to elucidate the relationship between acylation of flavonoids and the improvement of their bioactivity.

Objective To clarify the underlying mechanism of turmeric, which is traditionally used as a medicinal plant for the treatment of cardiovascular disorders, such as hypertension, and palpitations. Methods Methanol extracts of different turmeric were used. A tissue-organ-bath system was used to investigate the vasoactive effects of methanol extracts from 5 kinds of turmeric on isolated porcine basilar arteries. The arterial rings were suspended in physiological solution that was maintained at 37 °C temperature with a continuous supply of 95% O2 and 5% CO2. Results All turmeric extracts (20–800 μg/mL) induced concentration-dependent relaxation of the isolated porcine basilar artery pre-contracted with U46619 (1-5 × 10−9 M) in arterial rings with or without endothelium. There were no significant differences in the relaxation induced by different turmeric or between the endothelium-intact and denuded arteries. In depolarized, Ca2+-free medium, the turmeric extracts inhibited CaCl2-induced contractions and caused a concentration-dependent rightward shift of the response curves. In addition, propranolol (a non-specific β-adrenoceptor antagonist) slightly inhibited the relaxation induced by turmeric. In contrast, Nω-nitro-l-arginine, indomethacin, tetraethylammonium, glibenclamide and 4-aminopyridine did not affect turmeric-induced relaxation. Conclusion These results demonstrated that turmeric induced endothelium-independent relaxation of the porcine basilar artery, which may be due to the inhibition of extracellular and intracellular Ca2+ receptors and the partial inhibition of β-adrenergic receptors in vascular smooth muscle cells., Graphical abstract Image 1

6-(Methylsulfinyl)hexyl isothiocyanate (6-MSITC) is a major bioactive compound in Wasabi. Although 6-MSITC is reported to have cancer chemopreventive activities in rat model, the molecular mechanism is unclear. In this study, we investigated the anticancer mechanisms using two types of human colorectal cancer cells (HCT116 p53+/+ and p53−/−). 6-MSITC caused cell cycle arrest in G2/M phase and induced apoptosis in both types of cells in the same fashion. Signaling data revealed that the activation of ERK1/2, rather than p53, is recruited for 6-MSITC-induced apoptosis. 6-MSITC stimulated ERK1/2 phosphorylation, and then activated ERK1/2 signaling including ELK1 phosphorylation, and upregulation of C/EBP homologous protein (CHOP) and death receptor 5 (DR5). The MEK1/2 inhibitor U0126 blocked all of these molecular events induced by 6-MSITC, and enhanced the cell viability in both types of cells in the same manner. These results indicated that ERK1/2-mediated ELK1/CHOP/DR5 pathway is involved in 6-MSITC-induced apoptosis in colorectal cancer cells. Abbreviations: CHOP: C/EBP homologous protein; DR5: death receptor 5; ELK1: ETS transcription factor; ERK1/2: extracellular signal-regulated kinase 1/2; JNK: Jun-N-terminal kinase; MAPK: mitogen-activated protein kinase; MEK1/2: MAP/ERK kinase 1/2; 6-MSITC: 6-(methylsulfinyl)hexyl isothiocyanate; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP: poly(ADP-ribose) polymerase.

Objective: To assess the protective effects and mechanisms of cyanidin 3-glucoside (C3G) and its major phenolic metabolites including protocatechuic acid (PCA), phloroglucinaldehyde (PGA), vanillic acid (VA) and ferulic acid (FA) on non-alcoholic fatty liver disease (NAFLD). Design: First, C57BL/6J mice were fed a low-fat diet (LFD), high-fat diet (HFD) only, or HFD containing 0.4% C3G, PCA, PGA, VA or FA for 12 weeks to understand their protective effects against NAFLD. Next, the dose-dependent effect and mechanism of PCA against NAFLD were studied in a mouse model by supplementing 0.025% or 0.1% PCA in HFD. Eventually, the bacteriostatic effect of PCA on Enterococcus faecalis was confirmed in vitro, and the role of Enterococcus faecalis in NAFLD was discovered by fecal microbiota transplantation (FMT) experiment.Results: As the major phenolic metabolite of C3G, PCA dose-dependently reduced intraperitoneal and hepatic fat deposition, with alleviated insulin resistance, hepatic inflammation and lipid peroxidation. Characterization of gut microbiota indicated that PCA decreased the Firmicutes/Bacteroidetes ratio mainly by reducing the relative abundance of Roseburia, Intestinibacter and Enterococcus. Lipidomics and correlation analysis revealed that Enterococcus had a significant correlation with the changed lipid metabolites by PCA. In vitro bacteriostatic experiment showed that PCA has a direct inhibitory effect against Enterococcus faecalis, which has been proved to accelerate NAFLD by fecal microbiota transplantation.Conclusion: This study demonstrated the first time that PCA can protect against NAFLD through inhibiting Enterococcus faecalis.

Background Fisetin, a natural potent flavonoid, has various beneficial, pharmacological activities. In this study, we investigated expression changes of the fisetin regulating genes in lipopolysaccharide (LPS)-treated RAW264.7 cells and explored the role of fisetin in inflammation and autophagy. Methods and results Microarray analysis identified 1,071 genes that were regulated by fisetin in LPS-treated RAW264.7 cells, and these genes were mainly related to the process of immune system response. Quantitative real-time polymerase chain reaction and Bio-Plex analysis indicated that fisetin decreased the expression and secretion of several inflammatory cytokines in cells administered with LPS. Western blot analysis and immunofluorescence assay showed that fisetin decreased microtubule-associated protein 1 light-chain 3B (LC3B) and lysosome-associated membrane protein 1 (LAMP1) expression in LPS-treated cells, while the autophagy inhibitor chloroquine (CQ) could partially reverse this effect. In addition, fisetin reduced the elevated expression of p-PI3K, p-AKT and p-mTOR induced by LPS in a concentration-dependent manner. Conclusions Fisetin diminished the expression and secretion of inflammatory cytokines and facilitated autophagosome-lysosome fusion and degradation in LPS-treated RAW264.7 cells via inhibition of the PI3K/AKT/mTOR signaling pathway. Overall, the results of this study provide new clues for the anti-inflammatory mechanism of fisetin and explain the crosstalk between autophagy and inflammation to some extent.

Background Weaning is one of the major factors that cause stress and intestinal disease in piglets. Protocatechuic acid (PCA) is an active plant phenolic acid which exists in Chinese herb, Duzhong (Eucommia ulmoides Oliver), and is also considered as the main bioactive metabolite of polyphenol against oxidative stress and inflammation. This study aimed to investigate the effect of PCA on growth performance, intestinal barrier function, and gut microbiota in a weaned piglet model challenged with lipopolysaccharide (LPS). Methods Thirty-six piglets (Pig Improvement Company line 337 × C48, 28 d of age, 8.87 kg ± 0.11 kg BW) were randomly allocated into 3 treatments and fed with a basal diet (CTL), a diet added 50 mg/kg of aureomycin (AUR), or a diet supplemented with 4000 mg/kg of PCA, respectively. The piglets were challenged with LPS (10 μg/kg BW) on d 14 and d 21 by intraperitoneal injection during the 21-d experiment. Animals (n = 6 from each group) were sacrificed after being anesthetized by sodium pentobarbital at 2 h after the last injection of LPS. The serum was collected for antioxidant indices and inflammatory cytokines analysis, the ileum was harvested for detecting mRNA and protein levels of tight junction proteins by PCR and immunohistochemical staining, and the cecum chyme was collected for intestinal flora analysis using 16S rRNA gene sequencing. Results Dietary supplementation of PCA or AUR significantly increased the expression of tight junction proteins including ZO-1 and claudin-1 in intestinal mucosa, and decreased the serum levels of thiobarbituric acid reactive substances (TBARS) and IL-6, as compared with CTL group. In addition, PCA also decreased the serum levels of IL-2 and TNF-α (P P Prevotella 9, Prevotella 2, Holdemanella, and Ruminococcus torques group (P Roseburia and Desulfovibrio (P Conclusions These results demonstrated that both PCA and AUR had protective effect on oxidative stress, inflammation and intestinal barrier function in piglets challenged with LPS, and PCA potentially exerted the protective function by modulating intestinal flora in a way different from AUR. Holdemanella

Non-alcoholic fatty liver disease (NAFLD) is a disease that is prevalent worldwide, and its prevention by dietary administration has recently been considered as an important strategy. In this study, we administered mice with vine tea polyphenol (VTP) extracted from Ampelopsis grossedentata, a Chinese herb, to investigate the preventive effect on western diet (WD)-induced NAFLD. Male C57BL/6N mice were fed either a normal diet (ND) or WD with or without VTP for 12 weeks. The results revealed that VTP supplementation decreased the serum levels of cholesterol and triglycerides, and reduced the accumulation of hepatic lipid droplets caused by WD. Molecular data revealed that VTP enhanced fatty acid oxidation by reactivating the WD-suppressed phosphorylation of AMP-activated protein kinaseα (AMPKα) and the expressions of peroxisome proliferator-activated receptor alpha (PPARα), carnitine palmitoyl transferase IA (CPT1A) and cytochrome P450, family 4, subfamily a1 (CYP4A1). VTP inhibited hepatic lipogenesis by reducing the WD-enhanced level of mature sterol regulatory element-binding protein 1 (SREBP1) and fatty acid synthase (FAS). Moreover, VTP activated nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-mediated expressions of hemeoxygenase-1 (HO-1) and quinone oxidoreductase (NQO1), and reduced hepatic TBARS levels to prevent hepatic oxidative stress. On the other hand, VTP also increased intestinal zonula occludens-1 (ZO-1) expression and the relative abundance of gut Akkermansia, and reduced the ratio of Firmicutes/Bacteroidetes. Thus, VTP might prevent WD-induced NAFLD by balancing fatty acid oxidation and lipogenesis, hepatic oxidative stress, and gut microbiome, at least. These results suggest that vine tea, containing a high content of the bioactive compound dihydromyricetin, is a potential food resource for preventing NAFLD.

There are >80 species of turmeric (Curcuma spp.) and some species have multiple varieties, for example, Curcuma longa (C. longa) has 70 varieties. They could be different in their chemical properties and biological activities. Therefore, we compared antioxidant activity, total phenolic and flavonoid content of different species and varieties of turmeric namely C. longa [variety: Ryudai gold (RD) and Okinawa ukon], C. xanthorrhiza, C. aromatica, C. amada, and C. zedoaria. The antioxidant activity was determined using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, oxygen radical absorbance capacity (ORAC), reducing power and 2-deoxyribose (2-DR) oxidation assay. Our results suggested that RD contained significantly higher concentrations of total phenolic (157.4 mg gallic acid equivalent/g extract) and flavonoids (1089.5 mg rutin equivalent/g extract). RD also showed significantly higher DPPH radical-scavenging activity (IC50: 26.4 μg/mL), ORAC (14,090 μmol Trolox equivalent/g extract), reducing power absorbance (0.33) and hydroxyl radical scavenging activity (IC50: 7.4 μg/mL). Therefore, RD was chosen for the isolation of antioxidant compounds using silica gel column, Toyopearl HW-40F column, and high-performance liquid chromatography. Structural identification of the compounds was conducted using 1H NMR, 13C NMR, and liquid chromatography-tandem mass spectrometry. The purified antioxidant compounds were bisabolone-9-one (1), 4-methyllene-5-hydroxybisabola-2,10-diene-9-one (2), turmeronol B (3), 5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)-1-hepten-3-one (4), 3-hydroxy-1,7-bis(4-hydroxyphenyl)-6-hepten-1,5-dione (5), cyclobisdemethoxycurcumin (6), bisdemethoxycurcumin (7), demethoxycurcumin (8) and curcumin (9). The IC50 for DPPH radical-scavenging activity were 474, 621, 234, 29, 39, 257, 198, 47 and 18 μM and hydroxyl radical-scavenging activity were 25.1, 24.4, 20.2, 2.1, 5.1, 17.2, 7.2, 3.3 and 1.5 μM for compound 1, 2, 3, 4, 5, 6, 7, 8 and 9, respectively. Our findings suggested that the RD variety of C. longa, developed by the University of the Ryukyus, Okinawa, Japan, is a promising source of natural antioxidants.

Stress is a part of everyday life, but excessive stress can be related to diverse diseases. Recently, oral intake of a non-centrifuged cane sugar, Kokuto, was reported to produce potential anti-stress effects in humans. However, the molecular components associated with the anti-stress property of Kokuto remain mostly unknown. Therefore, we focused on the non-sugar component (NSC) fractions of Kokuto, and investigated how serum corticosterone level (used as a stress marker) and antioxidant activity were affected in restraint-stressed mice treated with NSC fractions obtained from the elusion on HP-20 resin with 25%, 50%, 75%, and 100% aqueous methanol (MeOH) solutions. Among the four NSC fractions, the 50% MeOH fraction showed a high content of phenolic compounds and high antioxidant activity. Moreover, oral administration of the 50% MeOH fraction suppressed both corticosterone secretion into the serum and reduction of antioxidant activity in serum and liver in restraint-stressed mice. Component analysis of the 50% MeOH fraction identified five antioxidative phenolic compounds: p-hydroxybenzaldehyde, p-hydroxyacetophenone, schaftoside, isoschaftoside, and p-coumaric acid. Phenolic compounds detected in the NSC fractions of Kokuto might contribute to the anti-stress property of Kokuto. In addition, this research provides more understanding of potential health benefits offered by the constituents of Kokuto.

The allicin diallyldisulfid-S-oxide, a major garlic organosulfur compound (OSC) in crushed garlic (Allium sativum L.), possesses antibacterial effects, and influences gut bacteria. In this study, we made allicin-free garlic (AFG) extract and investigated its effects on gut microbiome. C57BL/6N male mice were randomly divided into 6 groups and fed normal diet (ND) and high-fat diet (HFD) supplemented with or without AFG in concentrations of 1% and 5% for 11 weeks. The genomic DNAs of feces were used to identify the gut microbiome by sequencing 16S rRNA genes. The results revealed that the ratio of p-Firmicutes to p-Bacteroidetes increased by aging and HFD was reduced by AFG. In particular, the f-Lachnospiraceae, g-Akkermansia, and g-Lactobacillus decreased by aging and HFD was enhanced by AFG. The g-Dorea increased by aging and HFD decreased by AFG. In addition, the ratio of glutamic-pyruvic transaminase to glutamic-oxaloacetic transaminase (GPT/GOT) in serum was significantly increased in the HFD group and decreased by AFG. In summary, our data demonstrated that dietary intervention with AFG is a potential way to balance the gut microbiome disturbed by a high-fat diet.

Lotus seed is well known as traditional food and medicine, but its skin is usually discarded. Recent studies have shown that lotus seed skin contains a high concentration of proanthocyanidins that have multi-functions, such as antioxidation, anti-inflammation, and anti-cancer effects. In the present study, we aimed to isolate and purify the proanthocyanidins from lotus seed skin by acetone extraction and rotary evaporation, identify their chemical structures by HPLC-MS-MS and NMR, and further investigate the antioxidant properties of the extract purified by macroporous resin (PMR) from lotus seed skin both in vitro and in vivo. The results showed that PMR mainly contained oligomeric proanthocyanidins, especially dimeric procyanidin B1 (PB1), procyanidin B2 and procyanidin B4. Although it had limited ability to directly scavenge radicals in vitro, PMR could significantly enhance the expressions of antioxidant proteins via activation of nuclear factor-E2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in HepG2 cells. Molecular data revealed that PB1, a major component in PMR, stabilized Nrf2 by inhibiting the ubiquitination of Nrf2, which led to subsequent activation of the Nrf2-ARE pathway, including the enhancements of Nrf2 nuclear translocation, Nrf2-ARE binding and ARE transcriptional activity. Moreover, the in vivo results in high fat diet-induced mice further verified the powerful antioxidant property of PMR. These results revealed that lotus seed skin is a promising resource for functional food development.

Blue honeysuckle is rich in polyphenols, and recently receiving attention because of its potential antioxidant and anti-inflammatory properties. Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease that develops hepatic inflammation and metabolic syndrome. The present study aims to study the effect of blue honeysuckle extract (BHE) on fat deposition and hepatic lipid peroxidation in a high-fat-diet (HFD)-induced mouse model. Mice were fed a normal diet (ND) or a HFD containing 0.5% or 1% of BHE or not for 45 d. Liver sections were stained by hematoxylin-eosin staining. Serum lipids were measured by a clinical analyzer. Insulin was examined by ELISA, and hepatic proteins were detected by Western blotting. Dietary supplementation of BHE dose-dependently suppressed HFD-induced obesity and hepatic fat deposition. Moreover, BHE improved glucose metabolism by increasing insulin sensitivity and attenuated oxidative stress potentially by up-regulating nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-mediated pathway. Keywords: Blue honeysuckle, Polyphenols, Fatty liver, Lipid peroxidation, Antioxidant

L-theanine, a natural nonprotein amino acid with a high biological activity, is reported to exert anti-stress properties. An experiment with a 3 × 2 factorial arrangement was conducted to investigate the effects of dietary L-theanine on growth performance and immune function in lipopolysaccharide (LPS)-challenged broilers. A total of 432 one-day-old male yellow-feathered broilers were randomly assigned to 3 dietary treatments (control, antibiotic and L-theanine diets) with 2 subgroups of each (6 replicate cages; 12 birds/cage). Birds from each subgroup of the 3 dietary treatments were intra-abdominally injected with the same amount of LPS or saline at 24, 25, 26 d of age. Both dietary L-theanine and antibiotic improved (P

Turmeric (Curcuma spp.) are rhizomatous perennial herbs with broad spectrum of pharmacological actions. There are more than 80 species of turmeric and 70 varieties/strains of Curcuma longa, which may have different chemical properties and biological activities. Hence, we compared the major active components (curcuminoides) and antifungal activity of three Curcuma longa strains (Ryudai gold (RD), Okinawa ukon, and BK2), C. xanthorrhiza, C. aromatica, C. amada, and C. zedoaria against Fusarium solani sensu lato (FSSL). The content of curcuminoides was determined by HPLC and the antifungal activity was measured by the diameter of colonies grown on Petri dish, microscopic observation, and CLSI microdilution methods. The BK2 turmeric contained highest concentration of curcumin, demethoxycurcumin, and bisdemethoxycurcumin followed by RD, C. xanthorrhiza, Okinawa ukon, and C. aromatica. These compounds were not detected in C. amada and C. zedoaria. All turmeric species inhibited fungal growth in a concentration-dependent manner. The order of IC50 against FSSL was RD (78 to 92 μg/ml) > BK2 (89 to 101 μg/ml) > C. xanthorrhiza (98 to 114 μg/ml) > C. aromatica (183 to 204 μg/ml) > C. amada (183 to 206 μg/ml) > Okinawa ukon (191 to 216 μg/ml) > C. zedoaria (354 to 385 μg/ml). The results showed a correlation between the antifungal activity and curcuminiods contents of turmeric. Curcumin itself showed marked antifungal activity against FSSL (IC50 = 23 to 25 μg/ml) followed by demethoxycurcumin (IC50 = 25 to 27 μg/ml), while the antifungal activity of bisdemethoxycurcumin was extremely low (IC50 = 216 to 238 μg/ml). However, C. amada and C. zedoaria had no curcuminoids but showed antifungal effects which indicated that other compounds could also inhibit the growth of FSSL. The obtained results demonstrated that turmeric species C. longa (strains Ryudai gold and BK2) and C. xanthorrhiza had higher content of curcuminoids and showed excellent antifungal activity against FSSL.

6-(Methylsulfinyl)hexyl isothiocyanate (6-MSITC), a major bioactive compound in Wasabi [Wasabia japonica (Miq.) Matsum.], has revealed the inhibitory effect on colon carcinogenesis in rat cancer model although the underlying mechanism is unclear. In this study, we used two types of human colorectal cancer cells (HCT116 p53+/+ and HCT116 p53-/- ) to investigate the anticancer activity and molecular mechanisms of 6-MSITC. Interestingly, 6-MSITC inhibited the cell proliferation in both types of cells with similar IC50 value although a light increase in the phosphorylation and accumulation of P53 protein was observed in HCT116 p53+/+ cells at 24 h after treatment. In addition, 6-MSITC increased the ratio of proapoptotic cells in both types of cells with the same fashion in a p53-independent manner. The data from mitochondrial analysis revealed that 6-MSITC enhanced the ratio of proapoptotic B-cell lymphoma-2-associated X protein/antiapoptotic myeloid cell leukemia 1, and sequentially caused mitochondrial membrane potential (ΔΨm ) loss, cytochrome c release, and caspase-3 activation in both types of cells. Taken together, Wasabi 6-MSITC induced apoptosis of human colorectal cancer cells in p53-independent mitochondrial dysfunction pathway. These findings suggest that 6-MSITC might be a potential agent for colon cancer chemoprevention although with p53 mutation. © 2018 BioFactors, 2018.

Magnolol rich in Magnolia officinalis is a bioactive polyphenolic compound. The aim of this study was to examine the effects of magnolol additive (MA) on growth performance, expression levels of antioxidant-related genes, and intestinal mucosal morphology of Linwu ducks aged from 49 to 70 days, comparing with that of an antibiotic additive (colistin sulfate [CS]). A total of 275, 49-day-old ducks were assigned to 5 groups with 5 cages of 11 ducks each and fed diets supplemented with 0, 100, 200 and 300 mg of MA/kg and 300 mg of CS/kg for 3 weeks, respectively. The results showed that the average daily body weight gain (ADG) was increased significantly in MA-fed groups (200 and 300 mg/kg), compared with the basal diet (BD) group (P

Sanguinarine is a bioactive compound as a quaternary benzophenanthridine alkaloid from plant of the Macleaya cordata, Papaveraceae family. The present study was conducted to investigate the effects of dietary sanguinarine supplementation on growth performance, serum biochemistry parameters, intestinal mucosal morphology and gut microbiome in yellow feathered broilers. Two hundred and seventy 1-d-old female broilers were randomly assigned to 3 treatments ① Basal diet (NG); ② Basal diet containing bacitracin methylene disalicylate (50mg/Kg diet) (ANT); ③ Basal diet containing sanguinarine (0.7 mg/ kg of feed) (SAG). The statistical results showed that dietary sanguinarine supplementation enhanced growth performance and decreased glucose, uric acid as well as urea nitrogen levels of broilers at 28d of age (P

Protein fermentation has an adverse effect on colonic health; high-quality proteins and reducing the protein level (protein restriction) can effectively decrease the amount of proteins flowing into the colon for microbial protein fermentation. The study is aimed at determining the effects of different protein sources on colonic microbial composition and barrier function in nursery pigs with a low protein diet. A total of 264 weaned pigs were randomly divided into 4 dietary groups and each group had 6 pens with 11 pigs. Four low protein, amino acid (AA)-supplemented diets containing either 4% Palbio 50 RD (P50), Soyppt-50% (S50), concentrated degossypolized cottonseed protein (CDCP), or fish meal (FM) were prepared, and all the diets had similar digestible energy (DE), crude protein content (CP, about 18%), and equal amount of standardized ileal digestible (SID) lysine, methionine, tryptophan, and threonine. After 28 days of feeding trial, CDCP decreased the Desulfovibrio abundance but increased the Parabacteroides abundance. S50 elevated the Bacteroides and CF231 abundance. P50 and FM reduced the Clostridium and Ruminococcus abundance. CDCP upregulated the Occludin mRNA expression and tended to increase the amount of mixed neutral-acidic mucins in the colon. FM and CDCP declined the serum DAO and endotoxin contents. S50 and CDCP decreased the levels of serum IL-1α, and P50 lowered the serum IL-8 content. We concluded that plant protein (CDCP and S50) had advantages over animal protein (P50 and FM) in maintaining the colonic health via the regulation of colonic microbiota and barrier function.

Dietary protein sources have the potential to affect the colon microbiome of piglets that will subsequently have a large impact on metabolic capabilities and hindgut health. This study explored the effects of different protein sources on the growth performance, diarrhea rate, apparent ileal digestibility (AID) of crude protein (CP), colonic mucin chemotypes, colonic microbiome, and microbial metabolites of piglets. Twenty-four piglets were randomly divided into four groups that received isoenergetic and isonitrogenous diets containing either Palbio 50 RD (P50), Soyppt-50% (S50), concentrated degossypolized cottonseed protein (CDCP), or fish meal (FM) as the sole protein source. The experimental diets did not affect the estimated daily gain (EDG), but P50 increased fecal score compared with S50 and CDCP. CDCP increased, but P50 reduced AID of CP in comparison to FM and S50. S50 and CDCP increased the amount of mixed neutral-acidic mucins relative to P50. Venn analysis identified unique OTUs in the P50 (13), CDCP (74), FM (39), and S50 (31) groups. The protein sources did not change the colonic bacterial richness or diversity. High Escherichia abundance in the P50 and FM, great abundant of Lactobacillus in the CDCP, and high Gemmiger abundance in the S50 were found. The CDCP tended to elevate valeric acid and branched chain fatty acid (BCFA) concentrations compared with the other diets. The P50 and FM groups had greater ammonia nitrogen and methylamine contents than the S50 and CDCP groups. There was a positive correlation between the Escherichia and ammonia nitrogen, the Lactobacillus and short chain fatty acid (SCFA), and a negative correlation between the Gemmige and BCFA. These findings suggested short-term feeding of different protein sources did not affect the piglets' growth, but P50 increased the diarrhea rate. Potential pathogenic bacteria and detrimental metabolites appeared in the colons of piglets fed P50 and FM, whereas, beneficial effects were conferred upon piglets fed CDCP and S50, thus indicating that available plant proteins (cotton seed, soy) added to the diets of piglets enhanced colon health by reducing protein fermentation.

Ampelopsis grossedentata, also called vine tea, has been used as a traditional beverage in China for centuries. Vine tea contains rich polyphenols and shows benefit to human health, but the chemical and antioxidant properties of vine tea polyphenols from different locations remain unclear. This study aims to investigate the chemical and antioxidant properties of vine tea from three major production areas in China including Guizhou, Hunan, and Guangxi Provinces. The highest amount of polyphenol from vine tea was extracted by 70% ethanol at 70 &deg, C for 40 min with ultrasonic treatment. The major compound in vine tea polyphenols (VTP) was determined as dihydromyricetin (DMY) by high-performance liquid chromatography (HPLC) and the content was estimated as 21.42%, 20.17%, and 16.47% of dry weight basis from Hunan, Guizhou, and Guangxi products, respectively. The antioxidant activities were investigated in vitro and in culture hepatic cells. VTP and DMY showed strong 1,1-Diphenyl-2-picrylhydrazyl free radical (DPPH) scavenging ability and high oxygen radical absorption capacity (ORAC) value in vitro. VTP and DMY also increased the level of nicotinamide adenine dinucleotide phosphate (NADPH):quinone oxidoreductase (NQO1) in HepG2 cells. Moreover, VTP and DMY enhanced the level of nuclear factor erythroid 2-related factor 2 (Nrf2) and reduced the level of Kelch-like ECH-associated protein 1 (Keap1). Taken together, our data demonstrated that the extraction of vine tea by 70% ethanol with ultrasonic treatment is a novel method to efficiently obtain components possessing stronger antioxidant activity. Furthermore, the results from the culture cells suggest that the bioactive component of vine tea might exert the antioxidant activity by activating the cellular Nrf2/Keap1 pathway.

Fisetin, a dietary flavonoid, is reported to have cellular antioxidant activity with an unclear mechanism. In this study, we investigated the effect of fisetin on the nuclear factor, erythroid 2-like 2 (Nrf2) signaling pathway in HepG2 cells to explore the cellular antioxidant mechanism. Fisetin upregulated the mRNA expression of heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), and NAD(P)H quinone oxidoreductase-1 (NQO1), and induced the protein of HO-1 but had no significant effect on the protein of GCLC, GCLM and NQO1. Moreover, nuclear accumulation of Nrf2 was clearly observed by immunofluorescence analysis and western blotting after fisetin treatment, and an enhanced luciferase activity of antioxidant response element (ARE)-regulated transactivation was obtained by dual-luciferase reporter gene assays. In addition, fisetin upregulated the protein level of Nrf2 and downregulated the protein level of Kelch-like ECH-associated protein 1 (Keap1). However, fisetin had no significant effect on Nrf2 mRNA expression. When protein synthesis was inhibited with cycloheximide (CHX), fisetin prolonged the half-life of Nrf2 from 15 min to 45 min. When blocking Nrf2 degradation with proteasome inhibitor MG132, ubiquitinated proteins were enhanced, and fisetin reduced ubiquitination of Nrf2. Taken together, fisetin translocated Nrf2 into the nucleus and upregulated the expression of downstream HO-1 gene by inhibiting the degradation of Nrf2 at the post-transcriptional level. These data provide the molecular mechanism to understand the cellular antioxidant activity of fisetin.

Background: Serum lipids and inflammatory cytokines are considered as potential therapeutic targets for nonalcoholic fatty liver disease, which has become the most common chronic liver disease in adolescents. Cyanidin 3-glucoside (C3G) is a natural anthocyanin occurs widely in plants, and has shown potential inhibitory effect on inflammatory cytokines in our previous studies. Methods: First, serum levels of inflammatory cytokines including TNF-α and MCP-1 were measured in 80 adolescents (9-15 years old) selected based on their body mass index and serum levels of lipids. Second, the protective effects of C3G and its metabolites including protocatechuic acid (PCA), phloroglucinaldehyde (PGA), ferulic acid, and vanillic acid on nonalcoholic fatty liver disease were measured in a mouse model in which C57BL/6N mice (n=4 for each group) were fed a normal diet, high fat diet only, or high fat diet containing 0.4% of C3G or the four metabolites for 3 months. Finally, the regulatory effects of C3G and its metabolites on inflammatory pathways were clarified in mouse macrophages. Results: Compared with serum levels of MCP-1 (273.13±16.89 pg/ml, n=16) and TNF-α (26.07±1.36 pg/ml) in normal adolescents, those who were obese with abnormal high serum LDL-c/HDL-c ratio (2.88±0.11) had higher MCP-1 (345.19±11.82 pg/ml, P P P Conclusion: The serum LDL-c/HDL-c ratio is closely associated with the inflammatory status in nonalcoholic fatty liver disease, and C3G can suppress the inflammation via MAPK/ERK pathway potentially through its metabolites PCA and PGA.

Rice, broken rice (BR), and full-fat rice bran (FFRB) from six different origins were analyzed for their chemical composition, nitrogen-corrected apparent metabolized energy (AMEn), and standardized amino acid digestibility (SIAAD) in 14-day-old and 28-day-old Arbor Acres broilers. Results showed broilers fed with rice and BR had a similar AMEn regardless of the rice and BR having different CP, EE, NDF, ADF, and ash content. FFRB containing significantly different CP, EE, NDF, ADFm and starch presented variable AMEn (p &lt, 0.05), suggesting that starch content in rice and its byproducts contributed most to the AMEn of broilers. The regression equation of AMEn = 14.312 − (0.198 × NDF) and AMEn = 6.491 + (0.103 × Starch) were feasible to integrally predict AMEn of broilers fed to rice and its byproducts. Moreover, 28-day-old broilers had higher SIAAD than 14-day-old ones. The SIAAD of rice were higher than BR and FFRB except for Met, Cys, Thr, and Tyr in 14-day-old broilers (p &lt, 0.05), and the SIAAD of His, Asp, and Ser in BR were higher than FFRB (p &lt, 0.05). In 28-day-old broilers, the SIAAD of Leu, Trp, Asp, Gly, and Pro of rice were still higher than BR and FFRB (p &lt, 0.05), but BR and FFRB had no significant differences (p &gt, 0.05). The regression equations to estimate the SIAAD of Thr, Lys, and Met were: Met = 81.46 + (0.578 × CP), Thr = 0.863 + (6.311 × CP), and Trp = 102.883 − (1.77 × CP), indicating that CP content in rice and its byproducts was likely a major factor for prediction of SIAAD.

6-MSITC and 6-MTITC are sulforaphane (SFN) analogs found in Japanese Wasabi. As we reported previously, Wasabi isothiocyanates (ITCs) are activators of Nrf2-antioxidant response element pathway, and also inhibitors of pro-inflammatory cyclooxygenase-2. This study is the first to assess the global changes in transcript levels by Wasabi ITCs, comparing with SFN, in HepG2 cells. We performed comparative gene expression profiling by treating HepG2 cells with ITCs, followed by DNA microarray analyses using HG-U133 plus 2.0 oligonucleotide array. Partial array data on selected gene products were confirmed by RT-PCR and Western blotting. Ingenuity Pathway Analysis (IPA) was used to identify functional subsets of genes and biologically significant network pathways. 6-MTITC showed the highest number of differentially altered (≥2 folds) gene expression, of which 114 genes were upregulated and 75 were downregulated. IPA revealed that Nrf2-mediated pathway, together with glutamate metabolism, is the common significantly modulated pathway across treatments. Interestingly, 6-MSITC exhibited the most potent effect toward Nrf2-mediated pathway. Our data suggest that 6-MSITC could exert chemopreventive role against cancer through its underlying antioxidant activity via the activation of Nrf2-mediated subsequent induction of cytoprotective genes.

6-(Methylsulfinyl)hexyl isothiocyanate (6-MSITC), 6-(methylthio)hexyl isothiocyanate (6-MTITC), and 4-(methylsulfinyl)butyl isothiocyanate (4-MSITC) are isothiocyanate (ITC) bioactive compounds from Japanese Wasabi. Previous in vivo studies highlighted the neuroprotective potential of ITCs since ITCs enhance the production of antioxidant-related enzymes. Thus, in this present study, a genome-wide DNA microarray analysis was designed to profile gene expression changes in a neuron cell line, IMR-32, stimulated by these ITCs. Among these ITCs, 6-MSITC caused the expression changes of most genes (263), of which 100 genes were upregulated and 163 genes were downregulated. Gene categorization showed that most of the differentially expressed genes are involved in oxidative stress response, and pathway analysis further revealed that Nrf2-mediated oxidative stress pathway is the top of the ITC-modulated signaling pathway. Finally, real-time polymerase chain reaction (PCR) and Western blotting confirmed the gene expression and protein products of the major targets by ITCs. Taken together, Wasabi-derived ITCs might target the Nrf2-mediated oxidative stress pathway to exert neuroprotective effects.

8-prenyl quercetin (PQ) is a typical prenylflavonoid distributed in plant foods. It shows higher potential bioactivity than its parent quercetin (Q) although the mechanisms are not fully understood. This study aims to clarify the anti-inflammatory effects and molecular mechanisms of PQ in cell and animal models, compared to Q.RAW264.7 cells were treated with PQ or Q to investigate the influence on the production of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and protein kinases by Western blotting. Nitric oxide (NO) and prostaglandin E2 (PGE2 ) were measured by the Griess method and ELISA, respectively. Cytokines were assayed by the multiplex technology. Mouse paw edema was induced by LPS. The results revealed that PQ had stronger inhibition on the production of iNOS, COX-2, NO, PGE2 , and 12 kinds of cytokines, than Q. PQ also showed in vivo anti-inflammatory effect by attenuating mouse paw edema. Molecular data revealed that PQ had no competitive binding to Toll-like receptor 4 with LPS, but directly targeted SEK1-JNK1/2 (where SEK is stress-activated protein kinase and JNK1/2 is Jun-N-terminal kinase 1/2) and MEK1-ERK1/2 (where ERK is extracellular signal regulated kinase).PQ as a potential inhibitor revealed anti-inflammatory effect in both cell and animal models at least by targeting SEK1-JNK1/2 and MEK1-ERK1/2.

The present study was conducted to investigate the effects of dietary Achyranthes bidentata polysaccharide (ABPS) supplementation on performance, immune response, intestinal mucosal morphology, and gut microbiome in yellow-feathered broilers challenged with Escherichia coli K88. A 2 &times, 2 factorial design was used for the trial. Two hundred and forty one-day-old female broilers were randomly assigned to four treatments: (1) negative-control broilers were fed by a basal diet and saline (NG), (2) positive-control broilers were fed by a basal diet and orally challenged with 108 CFU E. coli K88 (CNG), (3) ABP group broilers were fed by a basal diet containing ABPS (500 mg/kg of feed) and saline, (4) CABP group broilers were fed by a basal diet containing ABPS (500 mg/kg of feed) and orally challenged with 108 CFU E. coli K88. Growth performance, serum biochemical indexes, immune responses, intestinal mucosal morphology, and cecal microbial community structure were evaluated. The ABP group had greatest body weight (BW), average daily body weight gain (ADG), and intestinal villus height compared to other treatments (p &lt, 0.05). The CABP group had a higher villus height/crypt depth ratio (V/C) compared with other treatments (p &lt, 0.05). The expression levels of NF-&kappa, B were lower in the ABP group. The CNG group had higher Escherichia coli and Enterococcus contents in cecal samples compared to other treatments (p &lt, 0.05). Serum glucose, uric acid, TNF-&alpha, and Secretory Immunoglobulin A (S-IgA) levels were higher in broilers challenged with E. coli (p &lt, 0.001) than that with saline. Broilers challenged with E. coli had reduced taxa richness in the cecal samples. Sequencing of 16S rRNA genes in cecal samples revealed that a lower proportion of Firmicutes and a higher proportion of Proteobacteria were detected in the broilers challenged with E. coli. Compared with the controls, dietary ABPS supplementation increased serum total protein, albumin, and S-IgA levels, but decreased serum glucose, uric acid, and TNF-&alpha, levels in broilers (p &lt, 0.05). Diet supplemented with ABPS increased the Firmicutes/Bacteroidetes ratio and the abundance of Ruminococcaceae and Lachnospiraceae, and altered cecal microbiota community structure. These results suggest that ABPS can promote growth performance and improve intestinal morphology and microbiota community structure in broilers challenged with E. coli K88.

Blue honeysuckle (Lonicera caerulea L) is a (poly) phenol-rich edible berry. Based on the properties of phenolic compounds, we investigated the anti-inflammatory and antioxidant effects of blue honeysuckle extract (BHE) in adjuvant-induced arthritis (AIA) rat model and macrophage-like (RAW264.7) cell model. Oral administration of BHE in the range of 75–300 µg/g BW attenuated AIA symptom as reducing paw edema in rats. The serum levels of pro-inflammatory factors including tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, and nitric oxide (NO) were significantly reduced in BHE-fed rat. The production of inflammatory enzymes, inducible nitric oxide synthases (iNOS) and cyclooxygenase-2 (COX-2), in the spleen was also significantly suppressed. Moreover, serum transaminases including glutamic oxaloacetic transaminase (GOT), glutamate-pyruvate transaminase (GPT) and gamma-glutamyl transferase (GGT) were inhibited, and the antioxidant enzymes including superoxide dismutase (SOD) and glutathione peroxidase (GPx) were recovered. Finally, the major bioactive components in BHE were identified as (−)-epicatechin (EC) and cyanidin 3-glucoside (C3G), which were proven to suppress the expression of COX-2 and iNOS, accompanied by enhancing the level of Nrf2 and HO-1 in RAW264.7 cells. Taken together, our data demonstrated that BHE rich with EC and C3G attenuated rat AIA symptom with crosstalk of anti-inflammatory and antioxidant effects.

Delphinidin 3-sambubioside (Dp3-Sam), a Hibiscus anthocyanin, was isolated from the dried calices of Hibiscus sabdariffa L, which has been used for folk beverages and herbal medicine although the molecular mechanisms are poorly defined. Based on the properties of Dp3-Sam and the information of inflammatory processes, we investigated the anti-inflammatory activity and molecular mechanisms in both cell and animal models in the present study. In the cell model, Dp3-Sam and Delphinidin (Dp) reduced the levels of inflammatory mediators including iNOS, NO, IL-6, MCP-1, and TNF-α induced by LPS. Cellular signaling analysis revealed that Dp3-Sam and Dp downregulated NF-κB pathway and MEK1/2-ERK1/2 signaling. In animal model, Dp3-Sam and Dp reduced the production of IL-6, MCP-1 and TNF-α and attenuated mouse paw edema induced by LPS. Our in vitro and in vivo data demonstrated that Hibiscus Dp3-Sam possessed potential anti-inflammatory properties.

Lonicera caerulea L. berry polyphenols (LCBP) are considered as major components for bioactivity. This study aimed to clarify the molecular mechanisms by monitoring inflammatory and antioxidant mediator actions in lipopolysaccharide (LPS)-induced mouse paw edema and macrophage cell model. LCBP significantly attenuated LPS-induced paw edema (3.0 ± 0.1 to 2.8 ± 0.1 mm, P0.05) and reduced (P0.05) serum levels of monocyte chemotactic protein-1 (MCP-1, 100.9 ± 2.3 to 58.3 ± 14.5 ng/mL), interleukin (IL)-10 (1596.1 ± 424.3 to 709.7 ± 65.7 pg/mL), macrophage inflammatory protein (MIP)-1α (1761.9 ± 208.3 to 1369.1 ± 56.4 pg/mL), IL-6 (1262.8 ± 71.7 to 499.0 ± 67.1 pg/mL), IL-4 (93.3 ± 25.7 to 50.7 ± 12.5 pg/mL), IL-12(p-70) (580.4 ± 132.0 to 315.2 ± 35.1 pg/mL), and tumor necrosis factor-α (TNF-α, 2045.5 ± 264.9 to 1270.7 ± 158.6 pg/mL). Cell signaling analysis revealed that LCBP inhibited transforming growth factor β activated kinase-1 (TAK1)-mediated mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways, and enhanced the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and manganese-dependent superoxide dismutase (MnSOD) in earlier response. Moreover, cyanidin 3-glucoside (C3G) and (-)-epicatechin (EC), two major components of LCBP, directly bound to TAK1. These data demonstrated that LCBP might inhibit LPS-induced inflammation by modulating both inflammatory and antioxidant mediators.

Loquat (Eriobotrya japonica) leaves have been used as traditional medicine for chemoprevention and treatment of chronic inflammatory diseases although the evidence supporting their functions is still poor. This study aims to clarify the anti-inflammatory effects and molecular mechanisms of loquat tea extract (LTE), in both cell and animal models. LTE, especially C fraction, inhibited the production of pro-inflammatory factors including inducible nitric oxide synthase (iNOS), nitric oxide (NO), IL-6, RANTES and TNF-α. Cellular signaling data revealed that the downregulation of TGF-β-activated kinase (TAK1)-mediated both mitogen-activated protein kinase (MAPK) and NF-κB pathways were involved in the inhibition of pro-inflammatory factors by C fraction. Mouse paws edema model further confirmed the in vivo anti-inflammatory effects of LTE.

Oolong tea theasinensins are a group of tea polyphenols different from green tea catechins and black tea theaflavins, and they are considered as bioactive compounds in Oolong tea. In the present study, based on the properties of theasinensin and information about inflammatory processes, we investigated the anti-inflammatory activity and molecular mechanisms of theasinensin A (TSA) in both cell and animal models. In the cell model, TSA reduced the levels of pro-inflammatory mediators including inducible nitric oxide synthase (iNOS), nitric oxide (NO), interleukin-12 (IL-12) (p70), tumor necrosis factor alpha (TNF-α), and monocyte chemotactic protein-1 (MCP-1) induced by lipopolysaccharide (LPS). Cellular signaling analysis revealed that TSA downregulated MAPK/ERK kinase (MEK)-extracellular signal-regulated kinase (ERK) signaling. Pull-down assay and affinity data revealed that TSA might directly bind to MEK-ERK for the inhibitory action. In the animal model, TSA suppressed the production of IL-12 (p70), TNF-α, and MCP-1 and attenuated mouse paw edema induced by LPS.

Background : Fresh loquat leaves contain several kinds of flavonoids and have been reported to have preventive effects against some human diseases such as diabetes, coughs and ulcers,. Recently, fresh loquat leaves in Japan were processed to a beverage, called loquat tea, after the fresh leaves are roasted at 350C for 30 minutes. However, the scientific evidence supporting the functions of these processed leaves is still minimal. Objective : The aim of this study is to investigate the antioxidant and anti-inflammatory activities of roasted loquat tea extract (LTE) in vitro and in culture cells. Methods : Bioactive fractions of LTE were separated by column chromatograph. Antioxidant activities were determined by DPPH and ROS assay. Pro-inflammatory mediators cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) were determined by Western blot and ELISA assay, respectively. Chemical quantification and characterization were analyzed by HPLC, FR-IR, and NMR. Phenolic content was measured by Folin-Ciocalteu assay. Results : The results showed that loquat tea extract (LTE) possessed stronger DPPH scavenging activity than fresh. Cellular data revealed that LTE inhibited the production of reactive oxygen species (ROS), and further suppressed the production of COX-2 and PGE2 in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Chemical quantification and characterization data indicated that LTE contained new bioactive phenolic components that were produced from the roasting processes of fresh loquat leaves. Conclusions : Loquat tea made from roasted loquat leaves contained new bioactive phenolic compounds that contribute to its antioxidant and anti-inflammatory activities. Keywords : Loquat tea, Antioxidant activity, Anti-inflammatory activity, Chemical characterization

Scope Garlic is used for both culinary and medicinal purposes by many cultures. The garlic organosulfur compounds (GOSCs) are thought to be bioactive components. This study aims to clarify the antiinflammatory effects and molecular mechanisms of GOSCs in both cell and animal models. Methods and results RAW264.7 cells were treated with six kinds of GOSCs to screen their influence on cyclooxygenase-2 and inducible nitric oxide synthase expression by Western blotting. Prostaglandin E2 and nitrite were measured by ELISA and Griess reaction, respectively. Cytokines in culture medium were assayed by the multiplex technology. Proteins were detected by Western blotting. Mouse paw edema was induced by LPS. The results revealed that diallyl trisulfide (DATS) was a strongest inhibitor for cyclooxygenase and inducible nitric oxide synthase among GOSCs, and reduced the levels of LPS-induced IL-6, IL-10, IL-12(p70), KC, MCP-1, and TNF-α. Cellular signaling analysis revealed that DATS downregulated AKT1/TGF-β-activated kinase-mediated mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathways. Furthermore, DATS activated Nrf2-mediated expression of HO-1 and NQO1 and reduced LPS-induced intracellular reactive oxygen species, which may contribute to suppress inflammatory mediator production. Finally, in vivo data demonstrated that DATS attenuated LPS-induced mouse paw edema. Conclusion DATS as a potential inhibitor revealed antiinflammatory effect in both cell and animal models by downregulating AKT1/TGF-β-activated kinase-mediated NFκB and MAPK signaling pathways.

Scope 6-Methylthiohexyl isothiocyanate (6-MTITC), one of the major bioactive ingredients in Japanese Wasabi, has revealed cytoprotective and cancer chemopreventive effects. This study aims to clarify the molecular mechanisms how 6-MTITC modulates nuclear factor E2-related factor 2 (Nrf2)/Kelchlike ECH-associating protein 1 (Keap1) system in antioxidant-responsive element (ARE)-mediated nicotinamide adenine dinucleotide phosphate (NADP): quinone oxidoreductase 1 (NQO1) expression. Methods and results HepG2 cells were treated with 6-MTITC with varying time and dose. NQO1, Nrf2, and Keap1 proteins were detected by Western blotting. ARE transactivation was detected by electrophilic mobility shift assay and reporter gene assay. Nuclear localization of Nrf2 was determined by immunocytochemistry assay. Ubiquitination of Nrf2 and Keap1 was detected using immunoprecipitation after treatment with MG132. Small interfering RNA was used to knockdown Nrf2 or Keap1. The results revealed that 6-MTITC modulated Nrf2/ARE pathway by stimulating Keap1 modification, and inhibiting Nrf2 ubiquitination and protein turnover. These actions finally increased nuclear Nrf2 accumulation and ARE-binding activity. Moreover, silencing Nrf2 markedly reduced ARE-driven activity induced by 6-MTITC. Conclusion 6-MTITC modulated ARE-driven NQO1 expression by stabilizing Nrf2 with enhanced Keap1 modification and decreased Nrf2 degradation.

Scope Myricetin is a dietary flavonol and widely distributed in many edible plants. It has been reported to have many bioactivities and considered as a promising chemopreventive compound. The present study aimed to investigate the influences of myricetin on gene expressions in genome-wide and underlying mechanisms. Methods and results Among total 44K gene probes, myricetin treatment upregulated the signals of 143 gene probes (0.33% of total probes) and downregulated signals of 476 gene probes (1.08% of total probes) by greater than or equal to twofold in HepG2 cells. The network pathway analysis revealed that nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-mediated antioxidant response element (ARE) activation is involved in myricetin-induced genes expressions. Molecular data revealed that myricetin activated Nrf2-ARE pathway by inhibiting Nrf2 ubiquitination and protein turnover, stimulating Nrf2 expression and kelch-like erythroid cell-derived protein with CNC homology (ECH)-associated protein 1 modification. All of these events finally increased nuclear Nrf2 accumulation and ARE-binding activity to enhance ARE-mediated genes expressions. Additionally, treatment with Nrf2 small interfering RNA attenuated the myricetin-induced ARE activity and gene expression. Conclusion An Nrf2-mediated ARE activation is involved in myricetin-induced expression profiling in hepatic cells.

Ethnopharmacological relevance Baicalein is a flavone originally isolated from the roots of traditional Chinese medicinal herb, Scutellaria baicalensis, which has been proved as a promising chemopreventive compound for many chronic human diseases. Aim of the study The present study aimed to clarify the molecular mechanism targeted by baicalein. Materials and methods Gene expression profiling of HepG2 cells treated with baicalein was carried out, using the Affymetrix 42K oligonucleotide microarray in the present study. Microarray data analyzed by Ingenuity Pathway Analysis (IPA), further study performed by real time PCR, reporter gene assay, and Western blot. Results Among total 42K gene probes, baicalein treatment up-regulated the signals of 440 gene probes (1.04% of total gene probes) and down-regulated signals of 254 gene probes (0.6% of total gene probes) by ≥2-fold. These genes were categorized into 35 groups and hit for biological processes, molecular functions, and signaling pathways. The network and pathway analyses of these data further revealed that an Nrf2 (nuclear factor-erythroid 2 p45-related factor 2)-mediated ARE (antioxidant response element) pathway is involved in baicalein-induced gene expression of hepatic metabolic enzymes. The representative enzymes involved in Nrf2/ARE pathway were further confirmed at mRNA level by real time PCR and at protein level by Western blot analysis. Moreover, the ARE-reporter gene assay demonstrated that baicalein stimulated Nrf2-mediated ARE transactivation. Conclusions Our results provide a comprehensive data for understanding the hepatic metabolism, bioactive role and the molecular mechanisms of baicalein.

Purified polyphenolic-rich extracts from four native Australian fruits, Illawarra Plum ( Podocarpus elatus Endl., Podocarpaceae ), Kakadu Plum ( Terminalia ferdinandiana Exell, Combretaceae ), Muntries ( Kunzea pomifera F. Muell., Myrtaceae ) and Native Currant ( Acrotriche depressa R.Br., Epacridaceae ), were screened for their ability to modulate anti-inflammatory activity in LPS-activated murine macrophages (RAW 264.7). The Kakadu Plum extract inhibited the expression of inducible nitric oxide synthase (iNOS), and importantly was the only fruit in this study that displayed differential inhibition of the expression of cyclooxygenase (COX)-2 but not COX-1. Illawarra Plum (COX-2 and iNOS) and Native Currant (iNOS only) also inhibited inflammatory enzymes, while Muntries exhibited none of these activities under the same conditions. All evaluated extracts inhibited the production of prostaglandin E 2 and nitric oxide, suggesting the involvement of alternative pathways in their regulation for the Muntries extract. Further molecular investigations, showed that Kakadu Plum inhibited the NF-кB pathway, but not the p44/42 mitogen activated protein kinase (MAPK) pathway. Collectively, these results demonstrate potential anti-inflammatory activities of native Australian fruits, in particular Kakadu Plum, in LPS-activated murine macrophages, thus confirming the potential biological activities of these fruits.

Objective: To isolate and identify the antifungal compounds from Curcuma amada. Methods: The antifungal activity was measured by the diameter of colonies grown on Petri dish, microscopic observation, and CLSI microdilution methods. The antifungal compounds were isolated through bioactivity guided purification by using silica gel and high-performance liquid chromatography. Structural identification of the antifungal compounds was conducted using 1H NMR, 13C NMR, and liquid chromatography-tandem mass spectrometry. Results: The purified antifungal compounds were zederone and furanodienone. These two compounds showed dose-dependent antifungal activity against Fusarium solani sensu lato. The concentration required for 50% growth inhibition (IC50) of FSSL ranged from 115 to 129 μM and 82 to 91 μM for zederone and furanodienone, respectively. Conclusion: This study suggested that the isolated compounds from Curcuma amada could be promising natural antifungal agents to control the diseases caused by Fusarium solani sensu lato.

JAK1/STAT3 pathway has been suggested to play a role in cell transformation and carcinogenesis. In the present study, we found that myricetin (3, 3', 4', 5, 5', 7-hexahydroxyflavone), a typical flavonol existing in many fruits and vegetables, could directly bind to JAK1/STAT3 molecules to inhibit cell transformation in epidermal growth factor (EGF)-activated mouse JB6 P(+) cells. Colony assay revealed that myricetin had the strongest inhibitory effect on cell transformation among three flavonols including myricetin, quercetin and kaempferol. Molecular data revealed that myricetin inhibited DNA- binding and transcriptional activity of STAT3. Furthermore, myricetin inhibited the phosphorylation of STAT3 at Tyr705 and Ser727. Cellular signaling analyses revealed that EGF could induce the phosphorylation of Janus Kinase (JAK) 1, but not JAK2. Myricetin inhibited the phosphorylation of JAK1 and increased the autophosphorylation of EGF receptor (EGFR). Moreover, ex vivo and in vitro pull-down assay revealed that myricetin bound to JAK1 and STAT3, but not EGFR. Affinity data further demonstrated that myricetin had a higher affinity for JAK1 than STAT3. Thus, our data indicate that myricetin might directly target JAK1 to block cell transformation in mouse JB6 cells.

In the title compound, C25H20O12, commonly known as pentaacetylated quercetin, the benzene ring and one of its methoxy substituent groups is disordered (site occupancy ratio 0.523:0.427), with a dihedral angle between the major-disorder component and the benzene ring of the benzopyranone moiety of 10.8 (6)°. In the crystal, C—H...O hydrogen-bonding interactions give chains which extend alongb.

Much is known about the bioactive properties of green tea flavan-3-ol. However, very little work has been done to determine the properties of proanthocyanidins, another kind of polyphenols in green tea. In this study, we have investigated the anti-inflammatory effect of tea prodelphinidin B-4 3'-O-gallate (PDG) by demonstrating the inhibitory effects on cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated murine macrophage RAW264 cells. PDG caused a dose-dependent inhibition of COX-2 and iNOS at both mRNA and protein levels with the attendant decrease of prostaglandin E2 (PGE2) and nitric oxide (NO) production. Molecular data revealed that PDG downregulated NF-kappaB signaling pathway. Electrophoretic mobility shift assay (EMSA) showed that PDG reduced the binding complex of NF-kappaB-DNA in the promoter of COX-2 and iNOS. Immunochemical analysis revealed that PDG suppressed LPS-induced phosphorylation and degradation of IkappaBalpha, and subsequent nuclear translocation of p65. Consequently, PDG suppressed phosphorylation of IkappaB kinase alpha/beta (IKKalpha/beta) and TGF-beta-activated kinase (TAK1). Taken together, our data indicated that PDG is involved in the inhibition of COX-2 and iNOS via the downregulation of TAK1-NF-kappaB pathway, revealing partial molecular basis for the anti-inflammatory properties of tea PDG.

The hexane, ethyl acetate, chloroform, ethanol and water extracts of aerial parts of Varthemia, Varthemia iphionoides, were investigated for cytotoxic activity against human myelocytic leukemia (HL-60) cells; DPPH radical-scavenging activity; antioxidative activity in the linoleic acid system; reducing power; antibacterial activity; the contents of phenolic compounds. A pronounced cytotoxic effect on human leukemia (HL-60) cells was shown in the hexane, chloroform and ethanol extracts, with inhibition of 89.0, 68.4 and 62.3%, respectively, at a concentration of 200 microg extract/ml. High DPPH radical-scavenging activity, antioxidative activity in the linoleic acid system and reducing power were found in the water and ethanol extracts, and were correlated to the contents of phenolic compounds. Antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, Escherichia coli, Bacillus cereus and Salmonella enteritides was shown in the ethyl acetate and chloroform extracts. A compound responsible for the antibacterial activity was isolated from the ethyl acetate extract, and identified as 3-oxocostusic acid.

The effects of anthocyanidins, the aglycon nucleuses of anthocyanins widely occurring in reddish fruits and vegetables, on the expression of cyclooxygenase-2 (COX-2) were investigated in lipopolysaccharide (LPS)-activated murine macrophage RAW264 cells. Of five anthocyanidins, delphinidin and cyanidin inhibited LPS-induced COX-2 expression, but pelargonidin, peonidin and malvidin did not. The structure-activity relationship suggest that the ortho-dihydroxyphenyl structure of anthocyanidins on the B-ring appears to be related with the inhibitory actions. Delphinidin, the most potent inhibitor, caused a dose-dependent inhibition of COX-2 expression at both mRNA and protein levels. Western blotting analysis indicated that delphinidin inhibited the degradation of IkappaB-alpha, nuclear translocation of p65 and CCAAT/enhancer-binding protein (C/EBP)delta and phosphorylation of c-Jun, but not CRE-binding protein (CREB). Moreover, delphinidin suppressed the activations of mitogen-activated protein kinase (MAPK) including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 kinase. MAPK inhibitors (U0126 for MEK1/2, SB203580 for p38 kinase and SP600125 for JNK) specifically blocked LPS-induced COX-2 expression. Thus, our results demonstrated that LPS-induced COX-2 expression by activating MAPK pathways and delphinidin suppressed COX-2 by blocking MAPK-mediated pathways with the attendant activation of nuclear factor-kappaB (NF-kappaB), activator protein-1 (AP-1) and C/EBPdelta. These findings provide the first molecular basis that anthocyanidins with ortho-dihydroxyphenyl structure may have anti-inflammatory properties through the inhibition of MAPK-mediated COX-2 expression.

The effect of a koji (Aspergillus awamori mut.) extract on the caffeoylquinic acid derivatives purified from sweetpotato (Ipomoea batatas L.) leaves was examined to develop the mass production of caffeic acid. A koji extract hydrolyzed the caffeoylquinic acid derivatives, chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid and 3,4,5-tri-O-caffeoylquinic acid, to caffeic acid. Furthermore, the koji extract also converted the major polyphenolic components from sweetpotato, burdock (Arctium lappa L.), and mugwort (Artemisia indica var. maximowiczii) leaves to caffeic acid. These results suggest that the production of caffeic acid from plant resources containing caffeoylquinic acid derivatives is possible.

The extract from baked sweet potato (Ipomoea batatas Lam. cv. Koganesengan) showed potential cancer-preventing effects. The extract was partially fractionated to four fractions (I, II-a II-b, and III) by Sephadex G-25 gel chromatography. The cytotoxicity against human myelocytic leukemia HL-60 cells, the suppression of TPA-induced transformation in mouse skin JB6 C141 cells, the apoptosis inducing activity in HL-60 cells, and the scavenging capacity against DPPH radical were tested on the four fractions. Fractions II-a and III showed markedly strong radical scavenging effects on the DPPH radical, coinciding with the high content of total phenolic compounds in the fractions. Both of these fractions suppressed strongly the proliferation of HL-60 cells with apoptosis induction in a dose-dependent manner. Moreover, the two fractions markedly blocked TPA-induced cell transformation in the JB6 cell line. Taken together, these data suggest that the water extract from baked sweet potato had potential chemopreventive properties.