Amino acids stimulate Akt phosphorylation,and reduce IL‐8 production and NF‐κB activity in HepG2 liver cells

Hepatic insulin resistance and inflammatory cytokine production contribute to the manifestation of the metabolic syndrome. As amino acids have been implicated in modulating insulin signaling and inflammation, we have investigated the effects of glutamine, leucine and proline on markers of inflammation and insulin sensitivity in HepG2 liver cells. Cells were incubated with IL‐1β (5 ng/mL) to stimulate IL‐8 production. After 24 h, glutamine inhibited IL‐8 production significantly (p<0.05) at 2, 5 and 10 mM (to 82, 73 and 72% of control), whereas leucine reduced IL‐8 production significantly only at 10 mM (66%) and proline at 5 and 10 mM (71 and 52%). Glutamine, leucine and proline all reduced NF‐κB activity after 3 h of IL‐1β stimulation at 2, 5 and 10 mM (p<0.001). Insulin‐induced (1 nM) Akt phosphorylation was reduced in cells treated with tumour necrosis factor‐α (10 ng/mL) for 24 h, but was partly restored by simultaneous incubation with glutamine, leucine and proline (25 mM). Phosphorylation of glycogen synthase kinase‐3β was unaffected by insulin stimulation and amino acid treatment. Our results indicate that glutamine, leucine and proline attenuate IL‐8 production, probably through inhibition of NF‐κB, and that they increase Akt phosphorylation in HepG2 cells.     

Mung Bean Protein Hydrolysate Modulates the Immune Response Through NF‐κB Pathway in Lipopolysaccharide‐Stimulated RAW 264.7 Macrophages

The objective of this study was to evaluate the immunomodulatory activity of mung bean protein hydrolysate (MBPH) in lipopolysaccharide (LPS)‐induced RAW 264.7 cells and discuss the possible immune regulatory mechanism. MBPH was prepared by alcalase, trypsin, neutrase, and flavourzyme. The 3‐h alcalase‐hydrolyzed hydrolysate with a molecular weight less than 1,450 Da was selected for the immunological tests. Results showed that MBPH possessed strong suppressing activity to proinflammatory mediators in a dose‐dependent manner. Compared to the LPS alone group, MBPH (200 µg/mL) significantly reduced nitric oxide (NO), inducible nitric oxide synthase, interleukin (IL)‐6, and IL‐1β secretion levels by 52.6%, 53.2%, 48.4%, and 49.7%, respectively, in LPS‐induced macrophages. It also enhanced IL‐10 secretion from 789 to 3,678 pg/mL. MBPH blocked nuclear factor‐kappa B (NF‐κB) translocation in LPS‐induced macrophages through the prevention of IκBα phosphorylation, and this process further prevented p65 translocation into the nucleus. A possible mechanism of MBPH is that it regulated the expression of inflammatory factors via the NF‐κB pathway, thus inhibiting inflammatory reactions. The results suggested that MBPH is of application potential in the development of immunomodulatory functional food to ameliorate immunosuppression.     

The postprandial inflammatory response after ingestion of heated oils in obese persons is reduced by the presence of phenol compounds

Scope Heating during the process of cooking alters the chemical properties of foods and may affect subsequent postprandial inflammation. We tested the effects of four meals rich in different oils subjected to heating on the postprandial inflammatory metabolism of peripheral blood mononuclear cells (PBMCs). Methods and results Twenty obese participants received four breakfasts following a randomized crossover design, consisting of milk and muffins made with different oils (virgin olive oil (VOO), sunflower oil (SFO), and a mixture of seeds oil (SFO/canola oil) with added either dimethylpolysiloxane (SOD), or natural antioxidants from olive mill wastewater alperujo (phenols; SOP)), previously subjected to 20 heating cycles. Postprandial inflammatory status in PBMCs was assessed by the activation of nuclear NF‐κB, the concentration in cytoplasm of the NF‐κB inhibitor (IκB‐α), the mRNA levels of NF‐κB subunits and activators (p65, IKKβ, and IKKα) and other inflammatory molecules (TNF‐α, IL‐1β, IL‐6, MIF, and JNK), and lipopolysaccharide (LPS) levels. VOO and SOP breakfasts reduced NF‐κB activation, increased IκB‐α, and decreased LPS plasma concentration. SFO increased IKKα, IKKβ, p65, IL‐1b, IL‐6, MIF, and JNK mRNA levels, and plasma LPS Conclusion Oils rich in phenols, whether natural (VOO) or artificially added (SOP), reduce postprandial inflammation, compared with seed oil (sunflower).     

Carbonyl compounds methylglyoxal and glyoxal affect interleukin‐8 secretion in intestinal cells by superoxide anion generation and activation of MAPK p38

The carbonyl compounds methylglyoxal (MG) and glyoxal (GL) are reactive intermediates of glucose degradation pathways and capable of inducing cellular damage. Although immune‐stimulating activity has been investigated in endothelial cells, little is known about the signaling pathways of cytokine induction of these compounds in the intestine. Hence, we investigated the impact of mitogen‐activated protein kinases (MAPK) and nuclear factor kappa B (NF‐κB) on IL‐8 production by human intestinal cells (Caco‐2 and HT‐29) after stimulation by MG and GL. Both compounds induced a dose‐dependent enhancement of IL‐8 secretion in human intestinal cells. MAPK p38 and extracellular signal‐regulated kinase (ERK) were phosphorylated in these cells after having been stimulated by MG and GL. Furthermore, inhibitors of MAPK p38 (SB 203580 and 239063), ERK1/2 (PD 98059) and NF‐κB activation (SM‐7368 and SC‐514) reduced IL‐8 secretion. The most important mechanism by which MG and GL induced IL‐8 secretion was the generation of superoxide anions which was confirmed by the inhibition of the cytosolic NADPH oxidase with diphenyl iodonium (DPI) or by application of superoxide dismutase (SOD). Our data suggest that multiple pathways were simultaneously activated; however, superoxide dependent MAPK p38 activation seems to be the most dominant pathway for IL‐8 secretion in intestinal cells.     

Suppression of IL‐8 Release by Sweet Olive Ethanolic Extract and Compounds in WiDr Colon Adenocarcinoma Cells

Oxidative stress can stimulate the secretion of pro‐inflammatory cytokines. Interleukin‐8 (IL‐8) has been implicated in the pathogenesis of inflammatory bowel disease and the metastatic spread of colorectal cancer. The flowers of Osmanthus fragrans (sweet olive) are used to alleviate dysentery with blood in the bowel, as well as stomach ache and diarrhea. However, the evidence of their therapeutic effects on these symptoms remains unclear. In the present study, the protective effects of sweet olive flower ethanolic extract (OFE) against oxidative stress in WiDr cells was assessed by evaluating its 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical scavenging activity. In addition, cellular IL‐8 secretion was evaluated. Notably, high‐performance liquid chromatography showed verbascoside to be the primary constituent in OFE; it exhibited a DPPH scavenging activity with an IC₅₀ of 8.23 μg/mL. Moreover, OFE (1 to 100 μg/mL) showed a potent, dose‐dependent inhibitory effect on H₂O₂‐induced IL‐8 secretion in WiDr cells. Nine compounds were isolated from OFE based on a protective effect‐guided purification process. Of these compounds, 5 phenolic compounds—verbascoside, phillygenin, tyrosol, methyl 4‐hydroxycinnamate, and eutigoside A—reduced IL‐8 secretion at 10 μg/mL treatment concentrations. Further analysis showed that the anti‐inflammatory effects of OFE likely occurred via nuclear factor‐κB pathway inhibition, which attenuates IL‐8 secretion in cells. Collectively, these data suggest that OFE could be developed as an agent that suppresses IL‐8 secretion to treat chronic inflammatory diseases.     

Dietary Trans Fats Enhance Doxorubicin‐Induced Cardiotoxicity in Mice

This study investigated the combined effects of trans fat diet (TFD) and doxorubicin upon cardiac oxidative, inflammatory, and coagulatory stress. TFD increased trans fatty acid deposit in heart (P < 0.05), and decreased protein C and antithrombin‐III activities in circulation (P < 0.05). TFD plus doxorubicin treatment elevated activities of plasminogen activator inhibitor‐1, lactate dehydrogenase, and creatine phosphokinase (P < 0.05). This combination also raised xanthine oxidase activity, and enhanced cardiac levels of reactive oxygen species, interleukin (IL)‐6, IL‐10, tumor necrosis factor‐alpha, and monocyte chemoattractant protein‐1 than TFD or doxorubicin treatment alone (P < 0.05). TFD alone increased cardiac nuclear factor kappa B (NF‐κB) activity (P < 0.05), but failed to affect expression of NF‐κB and mitogen‐activated protein kinase (MAPK) (P > 0.05). Doxorubicin treatment alone augmented cardiac activity, mRNA expression, and protein production of NF‐κB and MAPK (P < 0.05). TFD plus doxorubicin treatment further upregulated cardiac expression of NF‐κB p65, p‐p38, and p‐ERK1/2 (P < 0.05). These findings suggest that TFD exacerbates doxorubicin‐induced cardiotoxicity.     

Apple extract induces increased epithelial resistance and claudin 4 expression in Caco-2 cells

BACKGROUND: The small intestinal epithelium functions both to absorb nutrients, and to provide a barrier between the outside, luminal, world and the human body. One of the passageways across the intestinal epithelium is paracellular diffusion, which is controlled by the properties of tight junction complexes. We used a differentiated Caco‐2 monolayer as a model for small intestinal epithelium to study the effect of crude apple extracts on paracellular permeability. RESULTS: Exposure of crude apple homogenate to the differentiated Caco‐2 cells increased the paracellular resistance, determined as trans‐epithelial electrical resistance (TEER). This increase was linearly related to the concentration of apple present. The TEER‐enhancing effect of apple extract was due to factors mainly present in the cortex, and the induction was not inhibited by protein kinase inhibitors. Apple‐induced resistance was accompanied by increased expression of several tight junction related genes, including claudin 4 (CLDN4). CONCLUSION: Crude apple extract induces a higher paracellular resistance in differentiated Caco‐2 cells. Future research will determine whether these results can be extrapolated to human small intestinal epithelia. Copyright © 2011 Society of Chemical Industry     

Sarcodon aspratus Extract Ameliorates Dextran Sulfate Sodium‐Induced Colitis in Mouse Colon and Mesenteric Lymph Nodes

Mushrooms have been previously investigated for their immune‐modulating and anti‐inflammatory properties. We examined whether the anti‐inflammatory properties of Sarcodon aspratus ethanol extract (SAE) could elicit protective effects against dextran sulfate sodium (DSS)‐induced colitis in vivo. Male C57/BL6 mice were randomly assigned to 1 of 4 treatment groups: control (CON; n = 8), DSS‐treated (DSS; n = 9), DSS+SAE at 50 mg/kg BW (SAE50; n = 8), and DSS+SAE at 200 mg/kg BW groups (SAE200; n = 9). DSS treatment induced significant weight loss, which was significantly recovered by SAE200. Although SAE did not affect DSS‐mediated reductions in colon length, it improved diarrhea and rectal bleeding induced by DSS. SAE at 200 mg/kg BW significantly attenuated IL‐6 and enhanced IL‐10 expression in mesenteric lymph nodes (MLN), and significantly reduced IL‐6 levels in splenocytes. SAE200 also significantly attenuated DSS‐induced increase in IL‐6 and IL‐1β, and reductions in IL‐10 in colon tissue. High levels of SAE were also observed to significantly decrease inflammatory COX‐2 expression that was upregulated by DSS in mice colon. These findings may have relevance for novel therapeutic strategies to mitigate inflammatory bowel disease‐relevant inflammatory responses, via the direct and indirect anti‐inflammatory activity of SAE. We also found that SAE harbors significant quantities of total fiber and β‐glucan, suggesting a possible role for these components in protection against DSS‐mediated colitis.     

Characterization,Anti‐Inflammatory and Antiproliferative Activities of Natural and Sulfonated Exo‐Polysaccharides from Streptococcus thermophilus ASCC 1275

Exo‐polysaccharides (EPS) isolated from Streptococcus thermophilus ASCC 1275 were sulfated (31%). High‐performance liquid chromatography identified that EPS was composed of mannose (30.19%), galactose (20.10%), glucose (18.05%), glucosamine (16.04%), galactosamine (9.06%), glucuronic acid (3.55%), and ribose (3.01%). Pro‐/anti‐inflammatory cytokine secretion ratios (IL‐1β/IL‐10, IL‐6/IL‐10, and TNF‐α/IL‐10) of lipopolysaccharide stimulated RAW 264.7 macrophages were significantly decreased by EPS and S.EPS treatments in a dose dependent manner. Furthermore, anti‐inflammatory activities of S.EPS improved 49.3% and 24.0% than those of EPS before or after LPS treatment. The reactive oxygen species were inhibited by EPS and S.EPS by 49.6% and 55.1% at 50 μg/mL, respectively. Inhibition activities of S.EPS on nitric oxide production were 12.9% and 55.4% higher than those of EPS at 10 and 50 μg/mL. Additionally, S.EPS exhibited stronger antiproliferative activity on Caco‐2 and HepG2 cells. Our results indicated that anti‐inflammatory and antiproliferative activities of EPS were significantly (P < 0.01) improved by sulfonation.     

Anti‐Diabetic Effects of Gynura Bicolor Aqueous Extract in Mice

The effects of Gynura bicolor aqueous extract (GAE) upon glycemic control, coagulation disorder, lipid accumulation, and glycative, oxidative, and inflammatory stresses in diabetic mice were investigated. Mice were treated with streptozotocin to induce type 1 diabetes. Diabetic mice were divided into four groups, consumed GAE at 0%, 0.25%, 0.5%, or 1%. Normal group consumed standard mouse basal diet. After 8‐week treatments, mice were sacrificed after overnight fasting. Results showed that GAE supplement at 0.5% and 1% decreased plasma glucose level and increased plasma insulin level. Diabetes lowered plasma level of protein C and anti‐thrombin III; and raised plasminogen activator inhibitor‐1 activity and fibrinogen level in plasma. GAE supplement at 0.5% and 1% reversed these alterations. Histological data, assayed by Oil Red O stain, indicated that GAE supplement decreased lipid accumulation in liver. GAE supplement at 0.5% and 1% reduced aldose reductase activity in heart and kidney; and lowered the levels of carboxymethyllysine and pentosidine in plasma and two organs. Diabetes decreased glutathione content, and increased reactive oxygen species, interleukin (IL)‐1β, IL‐6, and tumor necrosis factor‐α production in heart and kidney. GAE supplement at three test doses reversed these changes. Diabetes upregulated the mRNA expression of p38 and nuclear factor kappa (NF‐κ)B in heart and kidney. GAE supplement suppressed the mRNA expression of both p38 and NF‐κB. These novel findings suggest that Gynura bicolor is a potent functional food for diabetic prevention or alleviation.     

Gallic acid, a histone acetyltransferase inhibitor, suppresses β-amyloid neurotoxicity by inhibiting microglial-mediated neuroinflammation

Scope: We examined the biological effect of gallic acid (GA) as a nuclear factor (NF)‐κB acetyltransferase inhibitor on microglial‐mediated β‐amyloid neurotoxicity and restorative effects on the Aβ‐induced cognitive dysfunction. Methods and results: The protective effects of GA on the survival of neuronal cells were assessed with an MTT assay and a co‐culture system. For the co‐culture experiments, both BV‐2 and primary microglia cells were treated with GA prior to Aβ stimulation, and conditioned media were transferred to Neuro‐2A cells. The mRNA and protein levels of inflammatory cytokines in both microglia and Neuro‐2A cells were assessed with real‐time polymerase chain reaction and western blotting. Inhibition of nuclear factor kappa B (NF‐κB) acetylation with GA treatment resulted in reduced cytokine production in microglia cells and protection of neuronal cells from Aβ‐induced neurotoxicity. Furthermore, we observed a restorative effect of GA on Aβ‐induced cognitive dysfunction in mice with Y‐maze and passive avoidance tests. Finally, we found that GA treatment efficiently blocked neuronal cell death by downregulating the expression of cytokines and the in vivo levels of NF‐κB acetylation. Conclusion: These results suggest that selective inhibition of NF‐κB acetylation by the histone acetyltransferase inhibitor GA is a possible therapeutic approach for alleviating the inflammatory progression of Alzheimer disease.     

Microwave drying of ginger (Zingiber officinaleRoscoe) and its effects on polyphenolic content and antioxidant activity

The effects of microwave (MW) drying on the total polyphenol content (TPP) and antioxidant properties of ginger extract (column extraction using 50% ethanol at room temperature) were evaluated using the Folin–Ciocalteau's method, 2, 2′‐diphenyl‐1‐picryl‐hydrazyl assay, reducing power, antioxidant capacity and ferric reducing ability of plasma assay. The results were compared to that of the convective cross‐flow drying (CD) at 50 ± 4 °C. Extract yield, TPP content (59–80 mg GAE per g) of the MW‐dried samples increased with increase in MW power levels (PL 40, 60, 80 and 100), which might be as a result of MW energy causing breakdown of cellular constituents. The MW‐dried (PL100, 800W) extract showed the highest quantity (1.5 fold) in TPP, [6]‐gingerol content and antioxidant activity, when compared to the CD extract. Hence, it might be concluded that the optimal MW PL for drying of ginger slices with respect to retention of nonvolatiles such as TPP including [6]‐gingerol was PL 100.     

Reverse Effect of Opuntia ficus‐indica L. Juice and Seeds Aqueous Extract on Gastric Emptying and Small‐Bowel Motility in Rat

This study was conducted to compare the effects of juice and seeds on gastric emptying, small‐bowel motility and intestinal ion transport. Separate groups of rats were randomized to receive NaCl, increasing doses of juice (5, 10, and 20 mL/kg, b.w.) or seeds aqueous extract (100, 200, and 400 mg/kg, b.w.). Simultaneously, two other groups were received, the reference drugs; clonidine (1 mg/kg) and yohimbine (2 mg/kg). The charcoal meal was used as a suspension for gastrointestinal motility test. The purgative action of juice was confirmed using the loperamide (5 mg/kg, p.o.) induced constipation. To evaluate the antisecretory effect, we were used as a hypersecretion agent, the castor oil at the dose of 5 mL/kg. Compared to the control and standard groups, we were showed that the prickly pear has an opposite effect on small‐bowel motility and gastric emptying. Indeed, the juice at various doses has a laxative effect of gastrointestinal transit in healthy and constipated‐rats. However, the aqueous extract of the seeds leads to a reduction of motility in normal rats which gives it a remarkable antidiarrhoeal activity, a notable intestinal fluid accumulation decline and electrolyte concentrations reestablishment. Moreover, orally juice administered at different doses accelerated the stomach emptying time in contrast to the seeds aqueous extract. More importantly, a significant variation in the phytochemical constituents levels between juice and seeds was found. These findings confirm the reverse therapeutic effects of this fruit in the treatment of digestive disturbances such as difficulty stool evacuation and massive intestinal secretion, likewise, the gastric emptying process perturbation.