Inflammatory Marker Genes Research Articles

Effect of insoluble fibre on intestinal morphology and mRNA expression pattern of inflammatory, cell cycle and growth marker genes in a piglet model

The effects of insoluble dietary fibre differing in lignin content on intestinal morphology and mRNA expression was tested in an animal model of 48 weaned piglets. Engaged fibre sources were wheat bran (rich in cellulose and hemicellulose) and pollen from Chinese Masson pine (Pinus massoniana) (rich in lignin), respectively. The fibre sources were added to a basal diet as follows: no addition (control), 3.0% wheat bran, 1.27% pine pollen, and 2.55% pine pollen. The 12 animals of each feeding group were fed four experimental diets ad libitum for 37 days and were then slaughtered for retrieving tissue samples from stomach, jejunum, ileum, colon and mesenterial lymph nodes. Both fibre sources increased villus height of mucosa in jejunum (+10% on average) and ileum (+16% on average). Results of mRNA expression rates of inflammatory, cell cycle and growth marker genes (NFκB, TNFα, TGFβ, Caspase3, CDK4, IGF1) were specific to fibre source and tissue: wheat bran induced an up-regulation of NFκB in stomach and jejunum, as well as TNFα and TGFβ, and Caspase3 in jejunum. Pine pollen induced down regulation of NFκB, TNFα, TGFβ, Caspase3, CDK4 and IGF1 in the colon as well as up-regulation of NFκB and TGFβ in mesenterial lymph nodes. Finally, an overall data comparison based on a hierarchical cluster analysis showed a close relation between gene regulation in different gut sections and organs, as well as between small intestine morphology and zootechnical performance.

Relations of Inflammatory Biomarkers and Common Genetic Variants With Arterial Stiffness and Wave Reflection

Inflammation causes vascular dysfunction and perpetuates proatherosclerotic processes. We hypothesized that a broad panel of inflammatory biomarkers and single nucleotide polymorphisms in inflammatory genes is associated with vascular stiffness. We assessed 12 circulating inflammatory biomarkers (C-reactive protein, fibrinogen, interleukin-6, intercellular adhesion molecule-1, lipoprotein-associated phospholipase-A2 [mass and activity], monocyte chemoattractant protein-1, myeloperoxidase, CD40 ligand, osteoprotegerin, P-selectin, and tumor necrosis factor receptor-II) in relation to tonometry variables (central pulse pressure, mean arterial pressure, forward pressure wave, reflected pressure wave, carotid-femoral pulse wave velocity, and augmentation index) measured in 2409 Framingham Heart Study participants (mean age: 60 years; 55% women; 13% ethnic/racial minorities). Single nucleotide polymorphisms (n=2195) in 240 inflammatory candidate genes were related to tonometry measures in 1036 white individuals. In multivariable analyses, biomarkers explained <1% of any tonometry measure variance. Applying backward elimination, markers related to tonometry (P<0.01) were as follows: tumor necrosis factor receptor-II (inversely) with mean arterial pressure; C-reactive protein (positively) and lipoprotein-associated phospholipase-A2 (inversely) with reflected pressure wave; and interleukin-6 and osteoprotegerin (positively) with carotid-femoral pulse wave velocity. In genetic association analyses, lowest P values (false discovery rate <0.50) were observed for rs10509561 (FAS), P=6.6x10(-5) for central pulse pressure and rs11559271 (ITGB2), P=1.1x10(-4) for mean arterial pressure. These data demonstrate that, in a community-based sample, circulating inflammatory markers tumor necrosis factor receptor-II (mean arterial pressure), C-reactive protein, lipoprotein-associated phospholipase-A2 activity (reflected pressure wave), interleukin-6, and osteoprotegerin (carotid-femoral pulse wave velocity) were significantly but modestly associated with measures of arterial stiffness and wave reflection. Additional studies are needed to determine whether variation in inflammatory marker genes is associated with tonometry measures.

Cardiac hypertrophy is enhanced in PPAR -/- mice in response to chronic pressure overload

Peroxisome proliferator-activated receptor-alpha (PPARalpha) is a nuclear receptor regulating cardiac metabolism that also has anti-inflammatory properties. Since the activation of inflammatory signalling pathways is considered to be important in cardiac hypertrophy and fibrosis, it is anticipated that PPARalpha modulates cardiac remodelling. Accordingly, in this study the hypothesis was tested that the absence of PPARalpha aggravates the cardiac hypertrophic response to pressure overload. Male PPARalpha-/- and wild-type mice were subjected to transverse aortic constriction (TAC) for 28 days. TAC resulted in a more pronounced increase in ventricular weight and left ventricular (LV) wall thickness in PPARalpha-/- than in wild-type mice. Compared with sham-operated mice, TAC did not affect cardiac function in wild-type mice, but significantly depressed LV ejection fraction and LV contractility in PPARalpha-/- mice. Moreover, after TAC mRNA levels of hypertrophic (atrial natriuretic factor, alpha-skeletal actin), fibrotic (collagen 1, matrix metalloproteinase-2), and inflammatory (interleukin-6, tumour necrosis factor-alpha, cyclo-oxygenase-2) marker genes were higher in PPARalpha-/- than in wild-type mice. The mRNA levels of genes involved in fatty acid metabolism (long-chain acyl-CoA synthetase, hydroxyacyl-CoA dehydrogenase) were decreased in PPARalpha-/- mice, but were not further compromised by TAC. The present findings show that the absence of PPARalpha results in a more pronounced hypertrophic growth response and cardiac dysfunction that are associated with an enhanced expression of markers of inflammation and extracellular matrix remodelling. These findings indicate that PPARalpha exerts salutary effects during cardiac hypertrophy.

The influence of apple- and red-wine pomace rich diet on mRNA expression of inflammatory and apoptotic markers in different piglet organs

Flavan-3-ols are a class of flavonoids that are widely distributed in fruits and beverages including red wine and apples. Consumption of flavanoid-rich food has been shown to exhibit anti-microbial, anti-oxidative, anti-inflammatory, and immune-modulating effects. To test the nutritional effects of flavanols on mRNA gene-expression of inflammatory and apoptotic marker genes, piglets were given two flavanoids-rich feeding regimens: a low flavanoid standard diet (SD) was compared with diets enriched with 3·5% apple pomace (APD) or 3·5% red-wine pomace (RWPD). The influence on mRNA expression levels was investigated in different immunological active tissues and in the gastro-intestinal tract (GIT). The investigation took place from 1 week prior weaning to 19 days post weaning in 78 piglets. The expression of expressed marker genes was determinate by one-step quantitative real-time (qRT-PCR): TNFα, NFκB as pro-inflammatory; IL10, as anti-inflammatory; caspase 3 as apoptosis; cyclin D1 as cell cycle marker; and nucleosome component histon H3 as reference gene.The feeding regimens result in tissue individual regulation of mRNA gene expression in all investigated organs. It was discovered that there were significant differences between the applied diets and significant changes during feeding time curse. Both pomace treatments caused a significant up-regulation of all investigated genes in liver. The effect on mesenterial lymph nodes and spleen was not prominent. In the GIT, the treatment groups showed a inhibitory effects on gene expression mainly in stomach and jejunum (NFκB, cyclin D1 and caspase 3). In colon the trend of caspase 3 was positive with the greatest change in the RWPD group.In jejunum and stomach the cell cycle turn over was reduced, whereas in liver the cell turn over was highly accelerate. The influence on inflammatory marker gene expression is mainly relevant in stomach. It is presume that both flavanoid rich feeding regimens have the potential to modulate the mRNA expressions of inflammatory, proliferation and apoptotic marker genes in the GIT and piglet organs.

Down syndrome critical region protein 1 (DSCR1), a novel VEGF target gene that regulates expression of inflammatory markers on activated endothelial cells

AbstractWe conducted a genome-wide analysis of genes that are regulated by vascular endothelial growth factor (VEGF) in endothelial cells and identified DSCR1 to be most significantly induced. Consistent with an antagonistic function on calcineurin (CnA) signaling, expression of DSCR1 in endothelial cells blocked dephosphorylation, nuclear translocation, and activity of nuclear factor of activated T cell (NFAT), a transcription factor involved in mediating CnA signaling. DSCR1 was not only induced by VEGF, but also by other compounds activating CnA signaling, suggesting a more general role for DSCR1 in activated endothelial cells. Transient expression of DSCR1 attenuated inflammatory marker genes such as tissue factor (TF), E-selectin, and Cox-2, identifying a previously unknown regulatory role for DSCR1 in activated endothelial cells. In contrast, knock-down of endogenous DSCR1 increased NFAT activity and stimulated expression of inflammatory genes on activated endothelial cells. Thus, the negative regulatory feedback loop between DSCR1 and CnA signaling in endothelial cells identified may represent a potential molecular mechanism underlying the frequently transient expression of inflammatory genes following activation of endothelial cells.

Myocardial and renal vascular protection with eplerenone, enalapril and coadministration in the SHHF rat

Recent experimental studies suggest a direct role for aldosterone in cardiovascular pathophysiology. Moreover, the EPHESUS trial (Eplerenone Post-AMI Heart Failure Efficacy and Survival Study) established that addition of the selective aldosterone blocker eplerenone to optimal medical therapy reduces morbidity and mortality in patients with acute myocardial infarction complicated by left ventricular dysfunction and heart failure (N Engl J Med 348:1309–1321, 2003). The potential mechanisms by which aldosterone may contribute to heart failure and the cardioprotective effects of aldosterone blockade were evaluated using the selective aldosterone blocker, eplerenone, in spontaneously hypertensive heart failure rats (SHHF). Fourteen month old SHHF rats were randomized to receive eplerenone (100 mg/kg/day, EPL), enalapril (10mg/kg/day, ENAL), or eplerenone + enalapril (EPL + ENAL). EPL, ENAL and EPL + ENAL treatment attenuated functional and structural decline and protected target organs. Compared to age-matched Sprague-Dawley controls, untreated SHHF exhibited elevated systolic blood pressure (SBP) (192±6 vs. 135±2 mmHg). ENAL (163±6 mmHg) or EPL + ENAL (151±6 mmHg) significantly decreased SBP, whereas SBP was not significantly reduced with EPL treatment (188±7 mmHg) compared to untreated SHHF (192±6 mmHg). EPL+ENAL improved ejection fraction (53±2 vs. 33±2%) and significantly reduced left ventricular end-diastolic (0.63±0.03 vs. 0.97±0.06 mL) and left ventricular end-systolic volume (0.29±0.02 vs. 0.65±0.05 mL) compared to untreated SHHF. Cardiac and renal histopathology revealed severe vascular injury, necrosis, fibrosis and inflammatory lesions in untreated SHHF. EPL + ENAL treatment attenuated myocardial damage, albuminuria (257.3±62.8 vs. 408.4±84.2 mg/24hr) and myocardial and renal inflammatory marker gene expression compared to untreated SHHF. Circulating osteopontin levels were also markedly reduced (∼3-fold). These data indicate that cardiac and renal damage in this model is accompanied by an aldosterone-mediated inflammatory response and coadministration of EPL + ENAL attenuates vascular inflammation and injury and provides superior target organ protection. Recent experimental studies suggest a direct role for aldosterone in cardiovascular pathophysiology. Moreover, the EPHESUS trial (Eplerenone Post-AMI Heart Failure Efficacy and Survival Study) established that addition of the selective aldosterone blocker eplerenone to optimal medical therapy reduces morbidity and mortality in patients with acute myocardial infarction complicated by left ventricular dysfunction and heart failure (N Engl J Med 348:1309–1321, 2003). The potential mechanisms by which aldosterone may contribute to heart failure and the cardioprotective effects of aldosterone blockade were evaluated using the selective aldosterone blocker, eplerenone, in spontaneously hypertensive heart failure rats (SHHF). Fourteen month old SHHF rats were randomized to receive eplerenone (100 mg/kg/day, EPL), enalapril (10mg/kg/day, ENAL), or eplerenone + enalapril (EPL + ENAL). EPL, ENAL and EPL + ENAL treatment attenuated functional and structural decline and protected target organs. Compared to age-matched Sprague-Dawley controls, untreated SHHF exhibited elevated systolic blood pressure (SBP) (192±6 vs. 135±2 mmHg). ENAL (163±6 mmHg) or EPL + ENAL (151±6 mmHg) significantly decreased SBP, whereas SBP was not significantly reduced with EPL treatment (188±7 mmHg) compared to untreated SHHF (192±6 mmHg). EPL+ENAL improved ejection fraction (53±2 vs. 33±2%) and significantly reduced left ventricular end-diastolic (0.63±0.03 vs. 0.97±0.06 mL) and left ventricular end-systolic volume (0.29±0.02 vs. 0.65±0.05 mL) compared to untreated SHHF. Cardiac and renal histopathology revealed severe vascular injury, necrosis, fibrosis and inflammatory lesions in untreated SHHF. EPL + ENAL treatment attenuated myocardial damage, albuminuria (257.3±62.8 vs. 408.4±84.2 mg/24hr) and myocardial and renal inflammatory marker gene expression compared to untreated SHHF. Circulating osteopontin levels were also markedly reduced (∼3-fold). These data indicate that cardiac and renal damage in this model is accompanied by an aldosterone-mediated inflammatory response and coadministration of EPL + ENAL attenuates vascular inflammation and injury and provides superior target organ protection.

Localized insulin-like growth factor I delivery to enhance new bone formation

Insulin-like growth factor I (IGF I) exerts an important role during skeletal growth and bone formation. Therefore, its localized delivery appears attractive for the treatment of bone defects. To prolong IGF I delivery, we entrapped the protein into biodegradable poly(lactide-co-glycolide) microspheres (PLGA MS) and evaluated the potential of this delivery system for new bone formation in two defect models of ovine long bones, i.e., a 8-mm methaphyseal drill hole and a 10-mm segmental tibia defect. Administration of 100 μg of IGF I in PLGA MS resulted in new bone formation within 3 weeks in the drill hole and bridging of the segmental defect within 8 weeks. The observed increase of 12% newly formed bone in the drill hole defect after 3 weeks was substantial, compared to the measured morphometric bone-to-total area ratio of 31% bone in normal cancellous bone. Bone regeneration was further explored by measuring gene expression of typical markers for local mediators and growth factors by real-time polymerase chain reaction. Inflammation was reduced in presence of IGF I and this in vivo observation was corroborated in vitro by quantifying gene expression of inflammatory proteins and by assessing the activation of the NF-κB pathway, playing an important role in the regulation of inflammation. Administration of the IGF I delivery system downregulated inflammatory marker gene expression at the site of bone injury, induced new bone formation and reduced bone resorption, and resulted in bridging of 10-mm segmental tibial defects within 8 weeks.

Inflammatory and Epithelial Responses in Mouse Strains That Differ in Sensitivity to Hyperoxic Injury

The pulmonary response to various toxicants including bleomycin, ozone, ionizing radiation, and hyperoxia is highly variable among mouse strains. The current study tests the hypothesis that at a similar stage of injury, regardless of strain, expression of inflammatory cytokine and epithelial marker genes would be similar, indicating a common pathway of injury progression. Three strains of mice, C57B1/6J, 129/J, and C3H/HeJ, ranging from sensitive to resistant, were exposed to >95% O2 for varying times. Ribonuclease protection was used to quantify changes in cytokine mRNA. Despite differences in the kinetics, each strain demonstrated similar hyperoxia-induced changes in the abundance of interleukin (IL)-6, IL-4β, IL-3, and tumor neucrosis factor (TNF)-α. For each strain, death was accompanied by similar increases in cytokine mRNAs above steady-state control levels. Other inflammatory cytokines, including IL-lα, IL-4, and interferon (IFN)-γ, were unaltered in all strains at all times. In situ hybridization analysis of the epithelial markers, surfactant protein B (SPB), and clara cell secretory protein (CCSP) at the time of proinflammatory induction showed a similar pattern of expression in all strains. Increased SPB was detected in bronchiolar epithelium, while the number of type II cells expressing this message declined. Both the number of cells expressing CCSP as well as abundance per cell declined. These results suggest that although differences in acute sensitivity to hyperoxia exist between mouse strains, once initiated, acute epithelial cell injury and associated inflammatory changes follow the same, pattern in all strains.