Anti-inflammatory activities of mogrosides from Momordica grosvenori in murine macrophages and a murine ear edema model

Momordica grosvenori (Luo Han Guo), grown primarily in Guangxi province in China, has been traditionally used for thousands of years by the Chinese to make hot drinks for the treatment of sore throat and the removal of phlegm. The natural noncaloric sweetening triterpenoid glycosides (mogrosides) contained in the M. grosvenori fruits are also antioxidative, anticarcinogenic, and helpful in preventing diabetic complications. The aim of this study was to assess the anti-inflammatory properties of mogrosides in both murine macrophage RAW 264.7 cells and a murine ear edema model. The results indicate that mogrosides can inhibit inflammation induced by lipopolysaccharides (LPS) in RAW 264.7 cells by down-regulating the expression of key inflammatory genes iNOS, COX-2, and IL-6 and up-regulating some inflammation protective genes such as PARP1, BCL2l1, TRP53, and MAPK9. Similarly, in the murine ear edema model, 12-O-tetradecanoylphorbol-13-acetate-induced inflammation was inhibited by mogrosides by down-regulating COX-2 and IL-6 and up-regulating PARP1, BCL2l1, TRP53, MAPK9, and PPARδ gene expression. This study shows that the anticancer and antidiabetic effects of M. grosvenori may result in part from its anti-inflammatory activity.     

DATS reduces LPS-induced iNOS expression, NO production, oxidative stress, and NF-kappaB activation in RAW 264.7 macrophages

Diallyl trisulfide (DATS), diallyl sulfide (DAS), and diallyl disulfide (DADS) are the three major organosulfur compounds (OSCs) in garlic oil. In contrast to DADS and DATS, evidence of an anti-inflammatory effect of DATS is limited. In this study compares the efficacy of DATS with those of DAS and DADS on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in RAW 264.7 macrophages. The NO production in LPS-activated RAW 264.7 macrophages was suppressed by both DADS and DATS in a dose-dependent manner. At 100 muM, the nitrite levels of DADS- and DATS-treated cells were 57 and 34%, respectively, of cells treated with LPS alone. DAS, however, had no influence on NO production even at a concentration of 1 mM. Western blot and Northern blot assays showed that DADS and DATS but not DAS dose-dependently suppressed LPS-induced iNOS protein and mRNA expression in a pattern similar to that noted for NO production. LPS-induced cellular peroxide production was significantly inhibited by DADS and DATS (P < 0.05) but not by DAS. Electrophoresis mobility shift assays further indicated that DADS and DATS effectively inhibited the activation of NF-kappaB induced by LPS. Taken together, these results indicate that the differential efficacy of three major OSCs of garlic oil on suppression of iNOS expression and NO production is related to the number of sulfur atoms and is in the order DATS > DADS > DAS. The inhibitory effect of DATS on LPS-induced iNOS expression is likely attributed to its antioxidant potential to inhibit NF-kappaB activation.     

Modulation of cytokine secretion by garlic oil derivatives is associated with suppressed nitric oxide production in stimulated macrophages

We previously described that garlic oil derivatives differentially suppress the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in activated macrophages. In the present study, we investigated the effects of the garlic derivatives, diallyl sulfide (DAS), diallyl disulfide (DADS), and allyl methyl sulfide (AMS), on cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and the association between modulation of cytokines and inhibition of NO production was also assessed. The results indicated that these garlic compounds had different effects on the secretion of activated cytokines, including proinflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6, as well as the antiinflammatory, IL-10. DAS inhibited the production of all stimulated cytokines in a concentration-dependent manner, and the inhibition was closely associated with the suppression of NO and PGE(2) production. DADS repressed the production of stimulated TNF-alpha and IL-10 and increased the production of activated IL-1beta and, to a lesser extent, IL-6; but only the decreased IL-10 production was associated with DADS-induced NO inhibition. Yet, the DAS- and DADS-suppressed NO production was independent of TNF-alpha. AMS, on the other hand, slightly suppressed the stimulated TNF-alpha but enhanced IL-10 production, and such modulation was closely associated with the decrease in NO production.     

Pterostilbene induces apoptosis and cell cycle arrest in human gastric carcinoma cells

Pterostilbene, an active constituent of blueberries, is known to possess anti-inflammatory activity and also induces apoptosis in various types of cancer cells. Here, the effects of pterostilbene on cell viability in human gastric carcinoma AGS cells were investigated. This study demonstrated that pterostilbene was able to inhibit cell proliferation and induce apoptosis in a concentration- and time-dependent manner. Pterostilbene-induced cell death was characterized with changes in nuclear morphology, DNA fragmentation, and cell morphology. The molecular mechanism of pterostilbene-induced apoptosis was also investigated. The results show the caspase-2, -3, -8, and -9 are all activated by pterostilbene, together with cleavage of the downstream caspase-3 target DNA fragmentation factor (DFF-45) and poly(ADP-riobse) polymerase. Moreover, the results indicate that the Bcl-family of proteins, the mitochondrial pathway, and activation of the caspase cascade are responsible for pterostilbene-induced apoptosis. Pterostilbene markedly enhanced the expression of growth arrest DNA damage-inducible gene 45 and 153 (GADD45 and GADD153) in a time-dependent manner. Flow cytometric analysis indicated that pterostilbene blocked cell cycle progression at G1 phase in a dose- and time-dependent manner. Pterostilbene increased the p53, p21, p27, and p16 proteins and decreased levels of cyclin A, cyclin E, cyclin-dependent kinase 2 (Cdk2), Cdk4, and Cdk6, but the expression of cyclin D1 was not affected. Over a 24 h exposure to pterostilbene, the degree of phosphorylation of Rb was decreased after 6 h. In summary, pterostilbene induced apoptosis in AGS cells through activating the caspase cascade via the mitochondrial and Fas/FasL pathway, GADD expression, and by modifying cell cycle progress and changes in several cycle-regulating proteins. The induction of apoptosis by pterostilbene may provide a pivotal mechanism of the antitumor effects and for treatment of human gastric cancer.     

Probiotics prevent the development of 1,2-dimethylhydrazine (DMH)-induced colonic tumorigenesis through suppressed colonic mucosa cellular proliferation and increased stimulation of macrophages

Probiotics modulate immunity and inhibit colon carcinogenesis in experimental models, but these effects largely depend on the bacterial strain, and the precise mechanisms are not well understood. Therefore, we studied the effect of Bifidobacterium longum and/or Lactobacillus gasseri on the development of 1,2-dimethylhydrazine (DMH)-induced colonic precancerous lesions and tumors in mice while delineating the possible mechanisms involved. The results suggest that dietary consumption of probiotics (B. longum and L. gasseri) resulted in a significant inhibition of DMH-induced aberrant crypt foci (ACF) formation in male ICR mice. Long-term (24 weeks) dietary consumption of probiotics resulted in a reduction of colon tumor multiplicity and the size of the tumors. Administration of B. longum and L. gasseri suppressed the rate of colonic mucosa cellular proliferation in a manner correlating with the inhibition of tumor induction by DMH. In addition, the phagocytic activity of peritoneal macrophages was significantly increased in the DMH-treated mice that were fed various doses of B. longum, but not with L. gasseri or combined probiotics (B. longum + L. gasseri). We also found that L. gasseri significantly increased the proliferation of RAW264.7 macrophage cells through an increase in S phase DNA synthesis, which was related to the up-regulation of proliferating cell nuclear antigen (PCNA) and cyclin A. Taken together, these results demonstrate the in vivo chemopreventive efficacy and the immune stimulating mechanisms of dietary probiotics against DMH-induced colonic tumorigenesis.     

DHA对脂肪细胞周期及COX-2 mRNA表达的调节

以大鼠前体脂肪细胞原代单层培养为模型，分别以0(对照组)、40(低剂量组)和160 μmol/L(高剂量组)的二十二碳六烯酸(DHA)处理未分化的前体脂肪细胞(培养第1天)和分化的脂肪细胞(分化第1天)。采用流式细胞仪(FCM)检测细胞周期分布；逆转录-聚合酶链反应(RT-PCR)分析环氧合酶-2(COX-2 )mRNA表达情况，探讨DHA对脂肪细胞周期及基因表达的调节作用。结果显示，未分化的前体脂肪细胞经DHA作用48 h，各处理组前体脂肪细胞G1期百分比均呈上升趋势，但与对照无显著性差异；分化的脂肪细胞经DHA作用24 h，40 μmol/L组细胞内COX-2 mRNA的表达量显著下降，而160 μmol/L组则显著增加。以上结果表明，DHA对大鼠前体脂肪周期无明显的调节作用，而对脂肪细胞内COX-2 mRNA的调节具有明显的浓度依赖效应。     

Licochalcone a inhibits lipopolysaccharide-induced inflammatory response in vitro and in vivo

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), is known for its antimicrobial activity and its reported ability to inhibit cancer cell proliferation. In the present study, we found that Lico A exerted potent anti-inflammatory effects in in vitro and in vivo models induced by lipopolysaccharide (LPS). The concentrations of TNF-α, interleukin (IL)-6, and IL-1β in the culture supernatants of RAW 264.7 cells were determined at different time points following LPS administration. LPS (0.5 mg/kg) was instilled intranasally (i.n.) in phosphate-buffered saline to induce acute lung injury, and 24 h after LPS was given, bronchoalveolar lavage fluid was obtained to measure pro-inflammatory mediator and total cell counts. The phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) p65 protein was analyzed by Western blotting. Our results showed that Lico A significantly reduced the amount of inflammatory cells, the lung wet-to-dry weight (W/D) ratio, protein leakage, and myeloperoxidase activity and enhances oxidase dimutase activity in mice with LPS-induced acute lung injury (ALI). Enzyme-linked immunosorbent assay results indicated that Lico A can significantly down-regulate TNF-α, IL-6, and IL-1β levels in vitro and in vivo, and treatment with Lico A significantly attenuated alveolar wall thickening, alveolar hemorrhage, interstitial edema, and inflammatory cells infiltration in mice with ALI. In addition, we further demonstrated that Lico A exerts an anti-inflammation effect in an in vivo model of acute lung injury through suppression of NF-κB activation and p38/ERK MAPK signaling in a dose-dependent manner.     

Pterostilbene is more potent than resveratrol in preventing azoxymethane (AOM)-induced colon tumorigenesis via activation of the NF-E2-related factor 2 (Nrf2)-mediated antioxidant signaling pathway

Inflammatory bowel diseases have been a risk factor of colorectal cancer (CRC). The reactive oxygen species (ROS) generated by inflammatory cells create oxidative stress and contribute to neoplastic transformation, proliferation, and even metastasis. Previously, resveratrol (RS) and pterostilbene (PS) had been reported to prevent chemical-induced colon carcinogenesis by anti-inflammatory and pro-apoptotic properties. In this study, we investigated whether RS and PS could prevent the azoxymethane (AOM)-induced colon tumorigenesis via antioxidant action and to explore possible molecular mechanisms. Male BALB/c mice were injected with AOM (5 mg/kg of body weight) with or without RS or PS, and at the end of the protocol, all of the mice were euthanized and colons were analyzed. Administrations of PS can be more effective than RS in reducing AOM-induced formation of aberrant crypt foci (ACF), lymphoid nodules (LNs), and tumors. We also find that PS is functioning more effectively than RS to reduce nuclear factor-κB (NF-κB) activation by inhibiting the phosphorylation of protein kinase C-β2 (PKC-β2) and decreasing downstream target gene expression, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and aldose reductase (AR) in mouse colon stimulated by AOM. Moreover, administration of RS and PS for 6 weeks significantly enhanced expression of antioxidant enzymes, such as heme oxygenase-1 (HO-1) and glutathione reductase (GR), via activation of NF-E2-related factor 2 (Nrf2) signaling. When the above findings are taken together, they suggest that both stilbenes block cellular inflammation and oxidative stress through induction of HO-1 and GR, thereby preventing AOM-induced colon carcinogenesis. In comparison, PS was a more potent chemopreventive agent than RS for the prevention of colon cancer. This is also the first study to demonstrate that PS is a Nrf2 inducer and AR inhibitor in the AOM-treated colon carcinogenesis model.     

Purification and characterization of a rhamnose-binding chinook salmon roe lectin with antiproliferative activity toward tumor cells and nitric oxide-inducing activity toward murine macrophages

In this study, a rhamnose-binding lectin from the roe of chinook salmon (Oncorhynchus tshawytscha) was purified and characterized, and its biological activities were examined in several model systems. Chinook salmon roe lectin had a molecular mass of 30 kDa and agglutinated rabbit and bovine erythrocytes. The hemagglutination activity of the lectin was not affected by metal ions. The lectin was stable up to 70 °C and between pH 4 and pH 11. Chinook salmon roe lectin did not exert antifungal activity toward the fungal species tested and did not exhibit mitogenic response toward mouse splenocytes up to a concentration of 5 mg/mL. The lectin had selective antiproliferative activity toward human breast cancer MCF-7 cells and hepatoma Hep G2 cells. It also induced the production of nitric oxide from mouse peritoneal macrophages. This is the first report that demonstrates these biological activities from chinook salmon roe lectin.     

Growth and nutritional disorders of coffee cultivated in nutrient solutions with suppressed macronutrients

The objective was to evaluate the effect of omitting macronutrients in the nutrients solution on growth characteristics and nutritional status of coffee. The treatments were complete nutrients solutions and solutions with nutrient omission: N (nitrogen), P (phosphorus), K (potassium), Ca (calcium), Mg (magnesium) and S (sulfur). The experiment was carried out under greenhouse conditions with 3 replicates in a completely random design. Plant height, number of leaves per plant, stem diameter, relative chlorophyll index, photosynthesis rate, stomatal conductance, transpiration, carbon dioxide (CO₂) concentration, dry matter, content levels of macronutrients in plant aerial part and root system, and nutritional disorders were evaluated. Macronutrients suppression affected nutrients concentration in many plant parts, inducing the appearance of symptoms characteristic of each nutrient. The most limiting nutrients for coffee plants development were nitrogen and calcium, reflected in the lower dry matter accumulation and nitrogen the most required.     

Pterostilbene, a new agonist for the peroxisome proliferator-activated receptor alpha-isoform, lowers plasma lipoproteins and cholesterol in hypercholesterolemic hamsters

Resveratrol, a stilbenoid antioxidant found in grapes, wine, peanuts and other berries, has been reported to have hypolipidemic properties. We investigated whether resveratrol and its three analogues (pterostilbene, piceatannol, and resveratrol trimethyl ether) would activate the peroxisome proliferator-activated receptor alpha (PPARalpha) isoform. This nuclear receptor is proposed to mediate the activity of lipid-lowering drugs such as the fibrates. The four stilbenes were evaluated at 1, 10, 100, and 300 microM along with ciprofibrate (positive control), for the activation of endogenous PPARalpha in H4IIEC3 cells. Cells were transfected with a peroxisome proliferator response element-AB (rat fatty acyl CoA beta-oxidase response element)-luciferase gene reporter construct. Pterostilbene demonstrated the highest induction of PPARalpha showing 8- and 14-fold increases in luciferase activity at 100 and 300 microM, respectively, relative to the control. The maximal luciferase activity responses to pterostilbene were higher than those obtained with the hypolipidemic drug, ciprofibrate (33910 and 19460 relative luciferase units, respectively), at 100 microM. Hypercholesterolemic hamsters fed with pterostilbene at 25 ppm of the diet showed 29% lower plasma low density lipoprotein (LDL) cholesterol, 7% higher plasma high density lipoprotein (HDL) cholesterol, and 14% lower plasma glucose as compared to the control group. The LDL/HDL ratio was also statistically significantly lower for pterostilbene, as compared to results for the control animals, at this diet concentration. Results from in vitro studies showed that pterostilbene acts as a PPARalpha agonist and may be a more effective PPARalpha agonist and hypolipidemic agent than resveratrol. In vivo studies demonstrate that pterostilbene possesses lipid and glucose lowering effects.     

Anti-stress effects of Glycine tomentella Hayata in tilapia: inhibiting COX-2 expression and enhancing EPA synthesis in erythrocyte membrane and fish growth

The objective of this study was to elucidate the in vivo effects of the ethanol extract of wooly Glycine tomentella Hayata (GTE) root on tilapia to elucidate whether GTE has antistress activity. Tilapia as an animal model were fed with or without GTE, then injected with lipopolysaccharide (LPS) or ammonium chloride (NH(4)Cl). The tilapia were exposed to 100 mg/L of aqueous NH(4)Cl, and/or acute cold stress. Growth parameters of the tilapia were measured during the feeding trials. Tilapia injected with GTE (20 μg/g of fish), NH(4)Cl (100 μg/g of fish) and/or LPS (1 μg/g of fish) were then sampled 2 h poststimulation. GTE significantly inhibited cyclooxygenase-2 expression and hemoglobin (Hb) dimer formation (36 kDa). GTE also improved growth and blood viscosity and upregulated eicosapentaenoic acid content of erythrocytes. The in vivo results indicated that GTE (20 μg/g of fish) can be applied as a stress-tolerance enhancing agent for the aquaculture industry.     

Ion chromatographic determination of perchlorate in foods by on-line enrichment and suppressed conductivity detection

Systemic uptake of perchlorate anion, a rocket fuel component and potential thyroid function disruptor, by leafy vegetables and other crops grown in contaminated waters is a public health concern. A column-switching anion-exchange chromatographic method with suppressed conductivity detection, described in this paper, achieved a 3-6 microg/kg method limit of quantitation in analysis of the wet weight edible portion of cantaloupe, carrots, lettuce, and spinach samples with field-incurred perchlorate. A test portion was blended with dilute nitric acid, and the extract was filtered under vacuum. A portion of the measured filtrate was acidified to pH approximately 2 by addition of cation-exchange resin, 4 mL was passed through a graphitized carbon cleanup column, and an aliquot of a collected fraction was pushed through a short precolumn for anion extraction, enrichment, and injection onto the analytical column. Statistical comparison with determination by tandem mass spectrometry-ion chromatography analysis of untreated filtrate revealed that the difference between means was not significant at the 95% confidence level (P value > or = 0.12) for crops tested. In addition, the method was applied to cooked vegetables processed as baby food.     

Influence of cucumariosides upon intracellular [Ca2+]i and lysosomal activity of macrophages

Biological effects of the triterpene glycosides, cucumariosides A(2)-2 and A(7)-1 from the edible sea cucumber Cucumaria japonica and their aglycones were investigated using embryos of the sea urchin Strongylocentrotus nudus and the BALB/C line mouse peritoneal macrophages. Cucumariosides were highly cytotoxic in a sea urchin embryo development test with EC(50) values of 0.3 and 1.98 microg/mL, respectively. The aglycone was completely lacking in cytotoxicity. In subtoxic concentrations (0.001-0.1 microg/mL), cucumarioside A(2)-2 showed more then 2-fold stimulation of lysosomal activity and induced a rapid short-term increase in cytosolic Ca(2+) content in mouse macrophages. The maximal stimulatory effect was detected after 1-2 h of cultivation of cells with this glycoside. Cucumarioside A(7)-1 demonstrated more weak effects and even slightly inhibited lysosomal activity, while the aglycone was completely ineffective. At the toxic concentration (1 microg/mL), cucumarioside A(2)-2 induced the sharp irreversable increase of intracellular Ca(2+) concentration. We suggested that cucumariosides, especially A(2)-2, may act as Ca(2+) agonists due to their membranolytic properties.     

Chondroitin sulfate extracted from ascidian tunic inhibits phorbol ester-induced expression of Inflammatory factors VCAM-1 and COX-2 by blocking NF-kappaB activation in mouse skin

Inflammatory factors are known to play a key role in promoting tumorigenesis; therefore, it is a promising strategy to inhibit the inflammation for cancer prevention. The current study was performed to investigate the potential effects of chondroitin sulfate (CS) extracted from ascidian tunic on the expression of inflammatory factors induced by treatment with 12- O-tetradecanoylphorbol-13-acetate (TPA) and to elucidate the underlying molecular mechanism of CS action in mouse skin inflammation. TPA was topically applied to the shaven backs of ICR mice with or without CS (1 or 2 mg) for 4 h. The results demonstrated that CS suppressed TPA-induced edema and reduced the expression of cyclooxygenase-2, vascular cell adhesion molecule-1, and Akt signaling in mouse skin. These studies suggest that CS from ascidian tunic may be developed as an effective natural anti-inflammatory agent.     

Methionine sulfoximine suppressed the stimulation of dark carbon fixation by ammonium nutrition in wheat roots

To evaluate the role of NH₄ ⁺ assimilates in dark carbon fixation in roots in providing carbon skeletons expended for NH₄ ⁺ assimilation, the rate of dark carbon fixation in roots was measured using NaH¹⁴CO₃. The ¹⁴C-metabolites were analyzed in wheat (Triticum aestivum L.) plants grown in NH₄ ⁺ media for various periods of time with or without methionine sulfoximine (MSX) treatment. The dark carbon fixation rate in the roots of wheat plants that had been grown with NH₄ ⁺ for 1 d was approximately 6-fold higher than the rate in control roots. The stimulation of dark carbon fixation in NH₄ ⁺-grown plants, however, was not observed in MSX-treated roots. In the roots of NH₄ ⁺-grown plants, the concentration and ¹⁴C-Iabeling of acidic metabolites such as citrate and malate considerably decreased whereas those of basic metabolites, especially asparagine, increased noticeably. With MSX treatment, the incorporation of ¹⁴C into basic metabolites was negligible. In response to NH₄ ⁺, phosphoenolpyruvate carboxylase (PEPC) activity increased, and PEPC proteins accumulated in wheat roots. Neither activity nor amounts of PEPC in roots increased in the presence of MSX. These findings suggest that primary assimilation of NH₄ ⁺ in roots is essential for the stimulation of dark carbon fixation, which coincides with the increased activity of root PEPC, to sufficiently replenish carbon skeletons necessary for NH₄ ⁺ assimilation.     

Anti-inflammatory effects of resveratrol and oligostilbenes from Vitis thunbergii var. taiwaniana against lipopolysaccharide-induced arthritis

Vitis thunbergii Sieb. and Zucc. var. taiwaniana Lu is an endemic plant in Taiwan used as a dietary supplement for bone health. In this study, human chondrocytes were induced to produce COX-2, MMP-3, -13, and PGE(2) by LPS. An (18)F-FDG microPET imaging system was used to evaluate the anti-inflammatory arthritic effects in vivo. Six stilbenes, resveratrol (1), (+)-ε-viniferin (2), ampelopsin C (3), ampelopsin A (4), (-)-vitisin B (5), and (+)-vitisin A (6), were isolated from the stem part of V. thunbergii, which displayed the strongest PGE(2) inhibition. Among these compounds, 1 significantly decreased COX-2 activity, PGE(2), MMP-3, and -13 production in vitro, and (18)F-FDG uptake and serum PGE(2) in rabbits in vivo. Anti-inflammatory effects were enhanced through the combined usage of 1 and other oligostilbenes. Taken together, the synergistic effects of 1 and oligostilbenes resulted in stem part extracts with lower 1 content displaying the better anti-inflammatory arthritis effects.