Relationship between intestinal mucosal iNOS mRNA expression and intestinal bacterial translocation in cirrhotic rats

Abstract The expression of CD14+ macrophages, CD4+, CD8+ lymphocytes and mRNA of inducible nitric oxide synthase (iNOS) was investigated in the endometrium of repeat breeders with subclinical endometritis [experimental group (EXP), n = 10] and healthy [control group (CTRL), n = 10] cows. The cows were selected on the basis of repeat breeding (3 unsuccessful inseminations), clinical and cytological examinations (> 10% polymorphonuclear neutrophils in uterine smears obtained by cytobrush). From all the cows endometrial biopsies were collected and the presence of CD14+, CD4+ and CD8+ cells in the endometrium was evaluated immunohistochemically using semi quantitative counting method. The mRNA expression of iNOS was determined using reverse transcription-PCR. In general, there were no significant differences between EXP and CTRL groups in the expression of CD4+ and CD8 + lymphocytes in all endometrial structures. In contrast, we observed a higher number of CD14+ macrophages in repeat breeding group compared to the control cows, however, this difference was slightly pronounced. CD14+ cells were detectable only in the stratum compactum and stratum spongiosum. The statistically significant (p ≤ 0.05) higher expression of iNOS mRNA was measured in the cows with subclinical endometritis compared to the healthy animals. Our results suggest that the increased expression of CD14+ macrophages and iNOS mRNA may be associated with embryonal mortality in repeat breeding cows with subclinical endometritis.

Download Full-text

Regulatory Actions of Toll-Like Receptor 2 (TLR2) and TLR4 in Leishmania donovani Infection in the Liver

Infection and Immunity ◽

10.1128/iai.01468-12 ◽

2013 ◽

Vol 81 (7) ◽

pp. 2318-2326 ◽

Cited By ~ 24

Author(s):

Henry W. Murray ◽

Yunhua Zhang ◽

Yan Zhang ◽

Vanitha S. Raman ◽

Steven G. Reed ◽

...

Keyword(s):

Mrna Expression ◽

Leishmania Donovani ◽

Interleukin 10 ◽

Inos Mrna ◽

Toll Like Receptor ◽

Content Type ◽

Toll Like Receptor 2 ◽

Ifn Γ ◽

Low Dose Chemotherapy ◽

Oxide Synthase

ABSTRACTIn livers of susceptible but self-curing C57BL/6 mice, intracellularLeishmania donovaniinfection enhanced Toll-like receptor 4 (TLR4) and TLR2 gene expression. In the liver, infected TLR4−/−mice showed reduced gamma interferon (IFN-γ), tumor necrosis factor (TNF), and inducible nitric oxide synthase (iNOS) mRNA expression, higher-level and slowly resolving infection, delayed granuloma formation, and little response to low-dose chemotherapy; in serum, the ratio of IFN-γ to interleukin 10 (IL-10) activity was decreased by 50%. In contrast, in TLR2−/−mice, control of liver infection, parasite killing, and granuloma assembly were accelerated and chemotherapy's efficacy enhanced. In livers of infected TLR2−/−mice, mRNA expression was not increased for inflammatory cytokines or iNOS or decreased for IL-10; however, the serum IFN-γ/IL-10 ratio was increased 6.5-fold and minimal responses to IL-10 receptor blockade suggested downregulated IL-10. In established infection in wild-type mice, blockading TLR2 induced parasite killing and triggering TLR4 strengthened resistance and promoted chemotherapy's effect. Thus, in experimentalL. donovaniinfection in the liver, TLR4 signaling upregulates and TLR2 signaling downregulates macrophage antileishmanial activity, making both receptors potential therapeutic targets in visceral leishmaniasis for engagement (TLR4) or blockade (TLR2).

Download Full-text

PPAR-γ ligands modulate effects of LPS in stimulated rat synovial fibroblasts

AJP Cell Physiology ◽

10.1152/ajpcell.2002.282.1.c125 ◽

2002 ◽

Vol 282 (1) ◽

pp. C125-C133 ◽

Cited By ~ 57

Author(s):

Marie-Agnès Simonin ◽

Karim Bordji ◽

Sandrine Boyault ◽

Arnaud Bianchi ◽

Elvire Gouze ◽

...

Keyword(s):

Mrna Expression ◽

Constitutive Expression ◽

Synovial Fibroblasts ◽

Binding Activity ◽

Inos Mrna ◽

Peroxisome Proliferator ◽

Peroxisome Proliferator Activated Receptor ◽

Ppar Γ ◽

Tnf Α ◽

Cox 2

This work demonstrated the constitutive expression of peroxisome proliferator-activated receptor (PPAR)-γ and PPAR-α in rat synovial fibroblasts at both mRNA and protein levels. A decrease in PPAR-γ expression induced by 10 μg/ml lipopolysaccharide (LPS) was observed, whereas PPAR-α mRNA expression was not modified. 15-Deoxy-Δ12,14-prostaglandin J2(15d-PGJ2) dose-dependently decreased LPS-induced cyclooxygenase (COX)-2 (−80%) and inducible nitric oxide synthase (iNOS) mRNA expression (−80%), whereas troglitazone (10 μM) only inhibited iNOS mRNA expression (−50%). 15d-PGJ2 decreased LPS-induced interleukin (IL)-1β (−25%) and tumor necrosis factor (TNF)-α (−40%) expression. Interestingly, troglitazone strongly decreased TNF-α expression (−50%) but had no significant effect on IL-1β expression. 15d-PGJ2 was able to inhibit DNA-binding activity of both nuclear factor (NF)-κB and AP-1. Troglitazone had no effect on NF-κB activation and was shown to increase LPS-induced AP-1 activation. 15d-PGJ2 and troglitazone modulated the expression of LPS-induced iNOS, COX-2, and proinflammatory cytokines differently. Indeed, troglitazone seems to specifically target TNF-α and iNOS pathways. These results offer new insights in regard to the anti-inflammatory potential of the PPAR-γ ligands and underline different mechanisms of action of 15d-PGJ2 and troglitazone in synovial fibroblasts.

Download Full-text

Prolonged colonic epithelial hyporesponsiveness after colitis: role of inducible nitric oxide synthase

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1999.276.3.g703 ◽

1999 ◽

Vol 276 (3) ◽

pp. G703-G710 ◽

Cited By ~ 27

Author(s):

Samuel Asfaha ◽

Cameron J. Bell ◽

John L. Wallace ◽

Wallace K. MacNaughton

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Mrna Expression ◽

Inducible Nitric Oxide Synthase ◽

Mucosal Inflammation ◽

Inos Mrna ◽

Trinitrobenzenesulfonic Acid ◽

Ussing Chambers ◽

Oxide Synthase ◽

Inducible Nitric Oxide

Colonic epithelial secretion is an important host defense mechanism. We examined whether a bout of colitis would produce long-lasting changes in epithelial function that persisted after resolution of mucosal inflammation. Colitis was induced in rats with intracolonic trinitrobenzenesulfonic acid. Six weeks later, colonic damage and inducible nitric oxide synthase (iNOS) mRNA expression and activity were measured. Segments of distal colon were mounted in Ussing chambers for measurement of permeability and responsiveness to secretory stimuli. Basal electrolyte transport parameters and permeability were not different from untreated controls. Despite normal macroscopic and histological appearance, secretory responses to electrical field stimulation (EFS), isobutylmethylxanthine (IBMX), and carbachol were significantly depressed (by 60–70%) relative to controls. iNOS mRNA expression and enzyme activity were significantly elevated. Dexamethasone reversed epithelial hyporesponsiveness and significantly reduced iNOS mRNA expression. A selective iNOS inhibitor normalized the secretory responses to EFS and IBMX but not to carbachol. These data suggest that ongoing synthesis of nitric oxide by iNOS contributes to chronic suppression of epithelial secretory function after episodes of colitis.

Download Full-text