Gene Expression Of Inflammatory Mediators Research Articles

Responses of human dental pulp cells after application of a low-concentration bleaching gel to enamel

ObjectiveTo evaluate the effect of a 17.5% H2O2 gel on the odontoblastic differentiation capability of human dental pulp cells (HDPCs). DesignThe bleaching gel was applied for 45, 15 or 5min to enamel/dentine discs adapted to transwells, positioned over previously cultured HDPCs. In the control group, no treatment was performed on the discs. Immediately after samples were bleached, the cell viability (MTT assay) and death (Live/Dead assay) as well as the mRNA gene expression of inflammatory mediators (TNFα, IL-1β, IL-6, and COX-2; real-time PCR) were evaluated. The mRNA gene expression of odontoblastic markers (DMP-1, DSPP, and ALP) and mineralized nodule deposition (alizarin red) were assessed at 7, 14 and 21 days post-bleaching. The amount of H2O2 in contact with cells was quantified. Data were evaluated by Kruskal–Wallis and Mann–Whitney tests (α=5%). ResultsSignificant cell viability reduction and cell death were observed for bleached groups relative to control in a time-dependent fashion. Also, significant overexpression of all inflammatory mediators tested occurred in the 45- and 15-min groups. In the bleached groups, the expression of ALP, DMP-1, and DSPP and the deposition of mineralized nodules were reduced in comparison with those in the control group, at the initial periods (7 and 14 days). However, the 15- and 5-min groups reached values similar to those in the control group at the 21-day period. ConclusionsThe 17.5% H2O2 gel was cytotoxic to pulp cells; however, cells subjected to short-term bleaching are capable of expressing the odontoblastic phenotype over time.

Early inflammatory mediator gene expression in two models of traumatic brain injury: ex vivo cortical slice in mice and in vivo cortical impact in piglets.

BackgroundThe immunological response during the first 24 hours after traumatic brain injury (TBI) may be a critical therapeutic interval for limiting the secondary neuronal damage that is influenced by enhanced inflammatory mediator expression.MethodsTo gain further insight of the early injury response, we examined the expression of several inflammatory genes by real-time qPCR as a function of time or distance from injury in two distinct mammalian models: an ex vivo mouse cortical slice injury system and an in vivo piglet model of brain injury.ResultsInterleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), chemokine ligands 2 (CCL2), 3 (CCL3), 4 (CCL4), and prostaglandin-endoperoxide synthase 2 (PTGS2) mRNAs increased within 5 h after injury in mouse cortical slices. Chemokine and PTGS2 mRNAs remained elevated in slices at 24 h, whereas IL-1β and TNF-α expressions decreased from earlier peak levels. At 24 h after cortical injury in 1-month-old piglets, the expression of CCL2 mRNA was significantly increased in the lesion core and in the penumbra region. The expression of PTGS2, IL-1β, and TNF-α was variable among the piglets.ConclusionsThese in vitro and large animal models of cortical injury expand our understanding of the early timing and spread of the immunological response and can serve as preclinical systems to facilitate the discovery of therapeutic agents for TBI aimed at regulating inflammatory mediator expression.

Lowering the dietary omega-6: omega-3 does not hinder nonalcoholic fatty-liver disease development in a murine model

It is hypothesized that a high dietary n-6:n-3 (eg, 10-20:1) is partly responsible for the rise in obesity and related health ailments. However, no tightly controlled studies using high-fat diets differing in the n-6:n-3 have tested this hypothesis. The aim of the study was to determine the role that the dietary n-6:n-3 plays in non-alcoholic fatty-liver disease (NAFLD) and colitis development. We hypothesized that reducing the dietary n-6:n-3 would hinder the development of NAFLD and colitis. Male C57BL/6 J mice were fed high-fat diets, differing in the n-6:n-3 (1:1, 5:1, 10:1, 20:1), for 20 weeks. Gas chromatography–mass spectrometry was used to analyze the hepatic phospholipid arachidonic acid (AA):eicosapentaenoic acid and AA:docosahexaenoic acid. Hepatic metabolism, inflammatory signaling, macrophage polarization, gene expression of inflammatory mediators, oxidative and endoplasmic reticulum stress, and oxidative capacity were assessed as well as colonic inflammatory signaling, and gene expression of inflammatory mediators and tight-junction proteins. Although reducing the dietary n-6:n-3 lowered the hepatic phospholipid AA:eicosapentaenoic acid and AA:docosahexaenoic acid in a dose-dependent manner and mildly influenced inflammatory signaling, it did not significantly attenuate NAFLD development. Furthermore, the onset of NAFLD was not paired to colitis development or changes in tight-junction protein gene expression. In conclusion, reducing the dietary n-6:n-3 did not attenuate NAFLD progression; nor is it likely that colitis, or gut permeability, plays a role in NAFLD initiation in this model.

Effects of Chronic and Acute Grape Consumption on Gene Expression of Inflammatory Mediators in PBMC

Studies have demonstrated that polyphenol‐rich foods offer protection against chronic metabolic diseases caused by inflammation and oxidative stress. We investigated whether consumption of grapes alters expression of inflammatory pathway genes and transcription factors in a chronic and acute setting. In this double‐blind, crossover trial, 20 obese subjects (BMI 37.0 ± 9.9) were randomized to consume either 60 g of freeze‐dried whole grape powder (Grape) daily (297 mg polyphenols) or matched placebo (C) daily (0 mg polyphenols) for 4 weeks separated by a 2‐week washout. Acute study days at the beginning of each intervention consisted of a high fat, high carbohydrate meal challenge (1,036 kcal, 31 g protein, 107 g carbohydrate, 54 g fat) accompanied by Grape or C consumption. Peripheral blood mononuclear cells (PBMCs) were isolated from fasted blood on acute study days and at postprandial time points 1, 3, and 5 hr. Fasted blood PBMCs were isolated at the end of each chronic, 4‐week intervention. Gene expression was assessed using real‐time quantitative PCR. Relative expression levels of target genes were calculated using the comparative CT method following normalization to the endogenous reference gene GAPDH. Expression of TNFα, NF‐κB, and IκBα genes in PBMCs shows no significant differences for Grape vs. C after 4 weeks of chronic consumption. Preliminary data examining expression of the same genes in the acute setting are inconclusive. Initial results indicate that grape consumption does not impact gene expression in inflammatory pathways; however additional genes must be investigated. (Supported by the California Table Grape Commission and USDA CRIS CA‐S*‐NTR‐6316‐H)

Mo1786 Different Strains of H. pylori Induce Tolerogenic Dendritic Cells in Children

murine homolog of human enteropathogenic E. coli, mice develop a TH1-mediated colitis with the resulting pathology attributed to inflammatory monocytes recruited to the colon by the chemokine CCL2. Previous evidence demonstrates that C. rodentium-induced CCL2 expression occurs through an NF-κB mediated mechanism within colonic stromal cells, however due to the intimate contact that is formed between the intestinal epithelial cell (IEC) and C. rodentium, it is possible the IEC is also a significant source of CCL2. Thus, this study was designed to test the hypothesis that NF-κB-derived gene expression from intestinal epithelial cells is necessary for the exacerbation of colitis via the recruitment of inflammatory monocytes. A corollary hypothesis was that inflammatorymonocytes contribute to infectious colitis via NF-κB activation. To test these hypotheses, two different strains of mice in which classical NF-κB activation is defective via the deletion of IKKβ in either intestinal epithelial cells (IKKβΔIEC) or myeloid-derived cells (IKKβΔMye) were orally challenged with C. rodentium. There was a reduction in C. rodentium-induced colon mass, colonic histopathology, colonic inflammatory mediator gene expression, and neutrophil accumulation in IKKβΔIEC mice as compared to wild type controls at the peak of infection. We also observed reductions in C. rodentium-induced colonic pathology, which was accompanied by increased macrophage accumulation in IKKβΔMye mice at the height of the infection. Interestingly, both IKKβΔIEC and IKKβΔMye mice had significantly higher levels of tumor necrosis factor (TNF)-α, as well as inducible nitric oxide synthase (i.e., iNOS) gene expression within the colon prior to C. rodentium challenge. Although exact mechanisms that render deficient mice more resistant to C. rodentium-induced colonic pathology remain to be elucidated, our data suggest that the deletion of canonical NF-κB signaling within epithelial cells of the colon is sufficient to create an environment which is unfavorable for the colonization by non-invading pathogens, including C. rodentium.

Anti-inflammatory potential of carotenoid meso-zeaxanthin and its mode of action

Context: meso-Zeaxanthin (MZ) is a xanthophyll carotenoid with profound antioxidant activity.Objective: Oxidative stress plays a decisive role in numerous degenerative diseases including cancer. The present study evaluates anti-inflammatory effect of MZ.Materials and methods: Balb/c mice were treated with different doses of MZ (50 and 250 mg/kg b.wt, orally) 5 d before subcutaneous injection of carrageenan (1%), dextran (1%), and formalin (2%). Paw edema formation in MZ-treated and -untreated animals was measured using vernier calipers. Anti-inflammatory activity of MZ against lipopolysaccharide (LPS)-induced inflammatory model was studied by culturing macrophages in the presence and absence of LPS (5 μg/ml) and different concentrations of MZ (5, 10, and 25 μg/ml). After 24 h, the effect of MZ on pro-inflammatory cytokine levels in macrophages was analyzed by ELISA and its effect on various inflammatory genes was studied by RT-PCR.Results: MZ administration at different doses significantly (p < 0.001) inhibited paw edema induced by carrageenan, dextran, and formalin in mice. MZ also exhibited profound anti-inflammatory effect against LPS-induced inflammation in macrophages. Increased production of nitric oxide, C-reactive proteins, and various pro-inflammatory cytokines (TNF-α, interleukin-1β, and interleukin-6) in LPS-stimulated macrophages was significantly reduced by MZ treatment. Moreover, LPS-stimulated up-regulated mRNA expression of various inflammatory mediator genes like COX-2, TNF-α, and iNOS was down-regulated by MZ administration.Discussion and conclusion: MZ has potent anti-inflammatory effect which can be due to its down-regulated expression of various inflammatory mediator genes. Since cancer is considered as an inflammatory disease, the present study points towards the importance of MZ in chemo-preventive strategy.

Hepatoprotective Effect of L entinus edodes Mycelia Fermented Formulation against Alcoholic Liver Injury in Rats

The hepatoprotective effects of fermented black rice bran extracts (FF1 and FF2: black rice bran fermented by Lentinus edodes derived from mycelium supplemented with soybean or Hovenia dulcis) and their associated mechanisms were evaluated. In an in vitro experiment, FFs caused significant amelioration of the metabolic function of rat hepatocytes treated with NH4Cl. In addition, administration of FFs to rats with chronic liver injury induced by 12-week continual alcohol consumption resulted in significant restoration of body weight shrinkage, notable attenuation of excessive aspartate aminotransferase, alkaline phosphatase and endotoxin in serum, malondialdehyde in liver and the lactulose/mannitol ratio in urine. Furthermore, FF1 or FF2 also caused significant downregulation of gene expression of several critical inflammatory mediators (interleukin-6, tumor necrosis factor-alpha, cyclooxygenase-2 and inducible nitric oxide synthase). Histopathological findings also indicated that FFs reduced inflammation, necrosis and fatty infiltration in liver tissue. Taken together, FFs exert hepatoprotective effects through anti-inflammatory and anti-lipid peroxidative properties and regulation of intestinal permeability. Practical Applications In this study, mycelia fermentation was utilized as a feasible strategy for enhancing the hepatotherapeutic effect of black rice and herbs. The findings demonstrated that Hovenia dulcis exerts a synergistic protective effect on an alcoholic liver disease animal model and could provide a new effective potential approach to alleviate alcoholic liver diseases.

Indirect cytocompatibility of a low-concentration hydrogen peroxide bleaching gel to odontoblast-like cells.

To assess the initial cytotoxicity and the late phenotype marker expression of odontoblast-like cells (MDPC-23) subjected to less aggressive in-office bleaching therapies. A 17.5% hydrogen peroxide (H2O2) gel was applied for 45, 15 or 5 min to enamel/dentine discs adapted to trans-wells positioned over cultured MDPC-23 cells. No treatment was performed on the negative control. Immediately after bleaching, the cell viability, gene expression of inflammatory mediators and quantification of H2O2 diffusion were evaluated. The ALP activity, DSPP and DMP-1 gene expression and mineralized nodule deposition (MND) were assessed at 7, 14 or 21 days post-bleaching and analysed statistically with Mann-Whitney U-tests (α = 5%). H2O2 diffusion, proportional to treatment time, was observed in all bleached groups. Reductions of approximately 31%, 21% and 13% in cell viability were observed for the 45-, 15- and 5-min groups, respectively. This reduction was significant (P < 0.05) for the 45- and 15-min groups, which also presented significant (P < 0.05) over-expression of inflammatory mediators. The 45-min group was associated with significant (P < 0.05) reductions in DMP-1/DSPP expression at all periods, relative to control. The ALP activity and MND were reduced only in initial periods. The 15-min group had less intense reduction of all markers, with no difference to control at 21 days. The 17.5% H2O2 applied to tooth specimens for 5 min caused no alteration in the odontoblast-like cells. When this gel was applied for 45 or 15 min, a slight cytotoxicity, associated with alterations in phenotypic markers, was observed. However, cells were able to recover their functions up to 21 days post-bleaching.

Spinal cord injury induced neuropathic pain: Molecular targets and therapeutic approaches.

Neuropathic pain, especially that resulting from spinal cord injury, is a tremendous clinical challenge. A myriad of biological changes have been implicated in producing these pain states including cellular interactions, extracellular proteins, ion channel expression, and epigenetic influences. Physiological consequences of these changes are varied and include functional deficits and pain responses. Developing therapies that effectively address the cause of these symptoms require a deeper knowledge of alterations in the molecular pathways. Matrix metalloproteinases and tissue inhibitors of metalloproteinases are two promising therapeutic targets. Matrix metalloproteinases interact with and influence many of the studied pain pathways. Gene expression of ion channels and inflammatory mediators clearly contributes to neuropathic pain. Localized and time dependent targeting of these proteins could alleviate and even prevent neuropathic pain from developing. Current therapeutic options for neuropathic pain are limited primarily to analgesics targeting the opioid pathway. Therapies directed at molecular targets are highly desirable and in early stages of development. These include transplantation of exogenously engineered cell populations and targeted gene manipulation. This review describes specific molecular targets amenable to therapeutic intervention using currently available delivery systems.

Effects of a blend of essential oils and an enzyme combination on nutrient digestibility, ileum histology and expression of inflammatory mediators in weaned piglets

The aim of this study was to test the hypothesis of an improved gut environment of post-weaning piglets when administered a blend of essential oils (EO; thymol and cinnamaldehyde) and an enzyme combination (xylanase and β-glucanase (XB)) either alone or in combination. To assess the effect of dietary treatments, faecal nutrient digestibility and microbial counts, as well as ileum histology and gene expression of inflammatory mediators were evaluated. One hundred and ninety-two weaned piglets were allocated into four experimental treatments, and fed the basal diet (CTRL) either without or with EO, XB or their combination (EO+XB) for a 42-day period. The experiment concerning digestibility was designed with two periods (period I: days 15 to 21; period II: days 29 to 35) and the faeces were collected on days 20, 21, 34 and 35. On day 42, six piglets from each treatment were slaughtered. It was found that EO, XB and EO+XB supplementation did not affect (P>0.05) the growth performance of the piglets from days 0 to 42. Moreover, no dietary effect on faecal score was observed. Faecal digestibility of dry matter, organic matter, ash, dietary fibre, lipid, CP and NDF were increased from period I to period II (P<0.01 to P=0.06), while no effect (P>0.05) of EO, XB or their combination on the faecal digestibility was observed at both periods. Compared with the CTRL diet, dietary XB reduced the faecal Lactobacillus and Escherichia coli counts but increased the Lactobacillus to Coliforms ratio on day 42 (P=0.02, 0.03 and 0.03, respectively), and all the additives supplementations decreased the counts of faecal Coliforms on day 42 (P<0.01). XB supplementation increased the villus to crypt ratio (P=0.04) and reduced the mucosal macrophages number (P<0.01) in the ileum compared with the CTRL group, and dietary EO or EO+XB decreased the number of lymphatic follicles (P=0.01 and P<0.01, respectively) and mucosal macrophages (P=0.02 and P<0.01, respectively). In addition, the interleukin (IL)-1α was downregulated in piglets treated with EO+XB compared with the EO group (P=0.02). In conclusion, the administration of EO, XB or their combination was effective in improving ileum histology, and EO+XB supplementation might benefit the modulation of the expression of ileum inflammatory cytokines in piglets.

Myocardial ischemic preconditioning upregulated protein 1(Mipu1):zinc finger protein 667 - a multifunctional KRAB/C2H2 zinc finger protein.

Myocardial ischemic preconditioning upregulated protein 1 (Mipu1) is a newly discovered upregulated gene produced in rats during the myocardial ischemic preconditioning process. Mipu1 cDNA contains a 1824-base pair open reading frame and encodes a 608 amino acid protein with an N-terminal Krüppel-associated box (KRAB) domain and classical zinc finger C2H2 motifs in the C-terminus. Mipu1 protein is located in the cell nucleus. Recent studies found that Mipu1 has a protective effect on the ischemia-reperfusion injury of heart, brain, and other organs. As a nuclear factor, Mipu1 may perform its protective function through directly transcribing and repressing the expression of proapoptotic genes to repress cell apoptosis. In addition, Mipu1 also plays an important role in regulating the gene expression of downstream inflammatory mediators by inhibiting the activation of activator protein-1 and serum response element.

The low level laser therapy (LLLT) operating in 660nm reduce gene expression of inflammatory mediators in the experimental model of collagenase-induced rat tendinitis.

Tendinopathy is a common disease with a variety of treatments and therapies. Laser therapy appears as an alternative treatment. Here, we investigate the effects of laser irradiation in an experimental model of tendinitis induced by collagenase injection on rats' Achilles tendon, verifying its action in important inflammatory markers. Male Wistar rats were used and divided into five groups: control saline (C), non-treated tendinitis (NT) and tendinitis treated with sodium diclofenac (D) or laser (1J) and (3J). The tendinitis was induced by collagenase (100μg/tendon) on the Achilles tendon, which was removed for further analyses. The gene expression for COX-2; TNF-α; IL-6; and IL-10 (RT-PCR) was measured. The laser irradiation (660nm, 100mW, 3J) used in the treatment of the tendinitis induced by collagenase in Achilles tendon in rats was effective in the reduction of important pro-inflammatory markers such as IL-6 and TNF-α, becoming a promising tool for the treatment of tendon diseases.

Coffee and liver health.

Coffee is one of the most widely used beverages in the world. It includes a wide array of components that can have potential implications for health. Several epidemiological studies associate coffee consumption with a reduced incidence of various chronic diseases such as diabetes, cardiovascular diseases, and neurodegenerative diseases. Over the past 20 years, an increasing number of epidemiological and experimental studies have demonstrated the positive effects of coffee on chronic liver diseases. Coffee consumption has been inversely associated with the activity of liver enzymes in subjects at risk, including heavy drinkers. Coffee favours an improvement in hepatic steatosis and fibrosis, and a reduction in cirrhosis and the risk of hepatocellular carcinoma. The mechanisms of action through which it exerts its beneficial effects are not fully understood. Experimental studies show that coffee consumption reduces fat accumulation and collagen deposition in the liver and promotes antioxidant capacity through an increase in glutathione as well as modulation of the gene and protein expression of several inflammatory mediators. Animal and in vitro studies indicate that cafestol and kahweol, 2 diterpens, can operate by modulating multiple enzymes involved in the detoxification process of carcinogens causing hepatocellular carcinoma. It is unclear whether the benefits are significant enough to "treat" patients with chronic liver disease. While we await clarification, moderate daily unsweetened coffee use is a reasonable adjuvant to therapy for these patients.

Isoflavones inhibit poly(I:C)-induced serum, brain, and skin inflammatory mediators - relevance to chronic fatigue syndrome.

BackgroundChronic Fatigue Syndrome (CFS) is a neuroimmunoendocrine disease affecting about 1% of the US population, mostly women. It is characterized by debilitating fatigue for six or more months in the absence of cancer or other systemic diseases. Many CFS patients also have fibromyalgia and skin hypersensitivity that worsen with stress. Corticotropin-releasing hormone (CRH) and neurotensin (NT), secreted under stress, activate mast cells (MC) necessary for allergic reactions to release inflammatory mediators that could contribute to CFS symptoms.ObjectiveTo investigate the effect of isoflavones on the action of polyinosinic:polycytidylic acid (poly(I:C)), with or without swim stress, on mouse locomotor activity and inflammatory mediator expression, as well as on human MC activation.MethodsFemale C57BL/6 mice were randomly divided into four groups: (a) control/no-swim, (b) control/swim, (c) polyinosinic:polycytidylic acid (poly(I:C))/no swim, and (d) polyinosinic:polycytidylic acid (poly(I:C))/swim. Mice were provided with chow low or high in isoflavones for 2 weeks prior to ip injection with 20 mg/kg poly(I:C) followed or not by swim stress for 15 minutes. Locomotor activity was monitored overnight and animals were sacrificed the following day. Brain and skin gene expression, as well as serum levels, of inflammatory mediators were measured. Data were analyzed using the non-parametric Mann-Whitney U-test.ResultsPoly(I:C)-treated mice had decreased locomotor activity over 24 hours, and increased serum levels of TNF-α, IL-6, KC (IL-8/CXCL8 murine homolog), CCL2,3,4,5, CXCL10, as well as brain and skin gene expression of TNF, IL-6, KC (Cxcl1, IL8 murine homolog), CCL2, CCL4, CCL5 and CXCL10. Histidine decarboxylase (HDC) and NT expression were also increased, but only in the skin, over the same period. High isoflavone diet reversed these effects.ConclusionPoly(I:C) treatment decreased mouse locomotor activity and increased serum levels and brain and skin gene expression of inflammatory mediators. These effects were inhibited by isoflavones that may prove useful in CFS.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-014-0168-5) contains supplementary material, which is available to authorized users.

Mechanical stress is a pro-inflammatory stimulus in the gut: in vitro, in vivo and ex vivo evidence.

AimsInflammatory infiltrates and pro-inflammatory mediators are found increased in obstructive and functional bowel disorders, in which lumen distention is present. However, what caused the low level inflammation is not well known. We tested the hypothesis that lumen distention- associated mechanical stress may induce expression of specific inflammatory mediators in gut smooth muscle.MethodsStatic mechanical stretch (18% elongation) was applied in vitro in primary culture of rat colonic circular smooth muscle cells (RCCSMCs) with a Flexercell FX-4000 Tension Plus System. Mechanical distention in vivo was induced in rats with an obstruction band placed in the distal colon.ResultsIn the primary culture of RCCSMCs, we found that static stretch significantly induced mRNA expression of iNOS, IL-6, and MCP-1 in 3 hours by 6.0(±1.4), 2.5(±0.5), and 2.2(±0.5) fold (n = 6∼8, p<0.05), respectively. However, gene expression of TNF-α, IL-1β, and IL-8 was not significantly affected by mechanical stretch. In the in vivo model of colon obstruction, we found that gene expression of iNOS, IL-6, and MCP-1 is also significantly increased in a time-dependent manner in the mechanically distended proximal segment, but not in the sham controls or distal segments. The conditioned medium from the muscle strips of the stretched proximal segment, but not the distal segment or control, significantly induced translocation and phosphorylation of NF-κB p65. This treatment further increased mRNA expression of inflammatory mediators in the naïve cells. However, treatment of the conditioned medium from the proximal segment with neutralizing antibody against rat IL-6 significantly attenuated the activation of NF-κB and gene expression of inflammatory mediators.ConclusionsOur studies demonstrate that mechanical stress induces gene expression of inflammatory mediators i.e. iNOS, IL-6, and MCP-1 in colonic SMC. Further ex vivo study showed that mechanical stress functions as a pro-inflammatory stimulus in the gut.

678P - Increased Gene Expression of Cytokines Induced By H. Pylori Infection and Changes After Eradication Therapy

ABSTRACT Aim: Chronic inflammation resulting from H. pylori infection is a risk factor for gastric cancer development, while the eradication of this bacterium can prevent malignant transformation. So, the aim of this study was to evaluate the mRNA quantitative expression of cytokines genes and receptors (TNFA, IL1B, IL6, IL12A, IL2 and TGFBRII) in patients with chronic H. pylori-positive gastritis before and after eradication treatment. Methods: Relative quantification (RQ) of mRNA expression was performed using specific probes for the target genes and ACTB and GAPDH reference genes by qPCR (TaqMan® gene expression assay). A total of 40 biopsies from patients with chronic gastritis were evaluated [28 H. pylori-positive (Hp+) before and 2-3 months after treatment, and 12 H. pylori-negative (Hp-)]. Wilcoxon signed-rank test was used to compare the groups before and after therapy. Results: TGFBRII showed reduced gene expression (RQ Conclusions: H. pylori infection increases the gene expression of inflammatory mediators that regulates the chronic inflammation such as TNFA, IL1B, IL6 and IL12A, however the eradication of bacterium does not completely normalize the gene expression of these cytokines, although TNFA expression has been reduced. Disclosure: All authors have declared no conflicts of interest.

Characterizing the Vocal Fold Epithelial Response to Recovery following Phonation‐Induced Trauma at the Transcriptional Level

Objectives: It is unknown how acute episodes of phonotrauma affect the recovery of the vocal fold epithelial barrier. The epithelial barrier is characterized by specialized adhesive structures. Our objective was to evaluate the gene expression of junctional complex proteins and inflammatory mediators following phonation-induced trauma in an in vivo rabbit model. Methods: Sixty-five New Zealand white breeder rabbits were randomized to normal (n = 5), modal intensity (n = 30), or raised intensity (n = 30) phonation for 120 minutes. Larynges were harvested at 0 hours, 4 hours, 8 hours, 1 day, 3 days, or 7 days following phonation and compared with normal. Real-time polymerase chain reaction was used to evaluate mRNA expression of occludin, zonula occluden-1 (ZO-1), E-cadherin, β-catenin, interleukin 1β (IL-1β), cyclooxygenase-2 (COX-2), and transforming growth factor β-1 (TGFβ1). Results: Both phonation groups demonstrated a down-regulation of occludin between 8 hours and 1 day ( P &lt; .016), and ZO-1 levels at 3 days ( P = .08). β-catenin and E-cadherin demonstrated an inverse relationship at 8 hours in the modal phonation group ( P = .08). IL-1β, COX-2, and TGFβ1 levels peaked at 8 hours in the modal intensity group ( P = .08), with TGFβ1 peaking at day 1 in the raised intensity group ( P = .08). Conclusions: Results revealed a critical time point at 8 hours and 1 day in which most junctional complex and inflammatory markers decreased or peaked. Given the mechanical stresses that the vocal folds must withstand on a regular basis, time-dependent changes in barrier and inflammatory gene regulation are critical to understanding vocal fold homeostasis and tissue recovery.

LPS에 의해 자극된 RAW264.7 세포에 대한 계지가출부탕의 항염증활동

Objectives GyejigaChulBuTang (GCBT) is a prescription used to treat acute and chronic arthritis in Korea, China, and Japan. This study assessed the anti-inflammatory and anti-oxidant activities of GCBT on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Methods Raw264.7 cells were pretreated with or without GCBT for 1 hour prior to incubation with LPS. Anti-inflammatory activity of GCBT was evaluated with reference to gene expression and production levels of proinflammatory cytokines (<TEX>$TNF{\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6, GM-CSF and <TEX>$INF{\gamma}$</TEX>) and inflammatory mediators (iNOS, COX-2, NO and <TEX>$PGE_2$</TEX>). In addition, intracellular ROS generation and signal transduction of MAPK family, PI3K/Akt and <TEX>$I{\kappa}B{\alpha}/NF{\kappa}B$</TEX> was investigated. Results Prior treatment with GCBT inhibited elevation of <TEX>$TNF{\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6, GM-CSF, <TEX>$INF{\gamma}$</TEX>, NO and <TEX>$PGE_2$</TEX>, together with their cognate mRNAs in a dose-dependent manner. Intracellular ROS contents were similarly reduced. These effects were due to inhibition of LPS-induced phosphorylation of MAPK family, PI3K/Akt and <TEX>$I{\kappa}B{\alpha}$</TEX> as well as nuclear translocation of <TEX>$NF{\kappa}B$</TEX>. Conclusions GCBT suppresses pro-inflammatory mediators. GCBT has potential in the treatment of juvenile rheumatoid arthritis associated with inflammation.

Expression and contributions of TRPM7 and KCa2.3/SK3 channels to the increased migration and invasion of microglia in anti-inflammatory activation states.

Microglia rapidly respond to CNS injury and disease and can assume a spectrum of activation states. While changes in gene expression and production of inflammatory mediators have been extensively described after classical (LPS-induced) and alternative (IL4-induced) microglial activation, less is known about acquired de-activation in response to IL10. It is important to understand how microglial activation states affect their migration and invasion; crucial functions after injury and in the developing CNS. We reported that LPS-treated rat microglia migrate very poorly, while IL4-treated cells migrate and invade much better. Having discovered that the lamellum of migrating microglia contains a large ring of podosomes – microscopic structures that are thought to mediate adhesion, migration and invasion – we hypothesized that IL4 and IL10 would differentially affect podosome expression, gene induction, migration and invasion. Further, based on the enrichment of the KCa2.3/SK3 Ca2+-activated potassium channel in microglial podosomes, we predicted that it regulates migration and invasion. We found both similarities and differences in gene induction by IL4 and IL10 and, while both cytokines increased migration and invasion, only IL10 affected podosome expression. KCa2.3 currents were recorded in microglia under all three activation conditions and KCNN3 (KCa2.3) expression was similar. Surprisingly then, of three KCa2.3 inhibitors (apamin, tamapin, NS8593), only NS8593 abrogated the increased migration and invasion of IL4 and IL10-treated microglia (and invasion of unstimulated microglia). This discrepancy was explained by the observed block of TRPM7 currents in microglia by NS8593, which occurred under all three activation conditions. A similar inhibition of both migration and invasion was seen with a TRPM7 inhibitor (AA-861) that does not block KCa2.3 channels. Thus, we conclude that TRPM7 (not KCa2.3) contributes to the enhanced ability of microglia to migrate and invade when in anti-inflammatory states. This will be an important consideration in developing TRPM7 inhibitors for treating CNS injury.

Antioxidant and Anti-inflammatory Effects of Yam (Dioscorea batatas Decne.) on Azoxymethane-induced Colonic Aberrant Crypt Foci in F344 Rats.

Yam (Dioscorea batatas Decne.) has long been used as a health food and oriental folk medicine because of its nutritional fortification, tonic, anti-diarrheal, anti-inflammatory, antitussive, and expectorant effects. Reactive oxygen species (ROS), which are known to be implicated in a range of diseases, may be important progenitors of carcinogenesis. The aim of this study was to investigate the modulatory effect of yam on antioxidant status and inflammatory conditions during azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. We measured the formation of aberrant crypt foci (ACF), hemolysate antioxidant enzyme activities, colonic mucosal antioxidant enzyme gene expression, and colonic mucosal inflammatory mediator gene expression. The feeding of yam prior to carcinogenesis significantly inhibited AOM-induced colonic ACF formation. In yam-administered rats, erythrocyte levels of glutathione, glutathione peroxidase (GPx), and catalase were increased and colonic mucosal gene expression of Cu/Zn-superoxide dismutase (SOD), Mn-SOD, and GPx were up-regulated compared to the AOM group. Colonic mucosal gene expression of inflammatory mediators (i.e., nuclear factor kappaB, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor alpha, and interleukin-1beta) was suppressed by the yam-supplemented diet. These results suggest that yam could be very useful for the prevention of colon cancer, as they enhance the antioxidant defense system and modulate inflammatory mediators.