Reduced Serotonin Transporter Levels and Inflammation in the Midbrain Raphe of 12 Month Old APPswe/PSEN1dE9 Mice

Background: Although mood and sleep disturbances are nearly universal among patients with Alzheimer's disease (AD), brain structures involved in non-cognitive processing remain under characterized in terms of AD pathology. Objectives: This study was designed to evaluate hallmarks of AD pathology in the brainstem of the APPswe/PS1dE9 mouse model of familial AD. Methods: Fresh-frozen sections from female, 12 month old, transgenic and control B6C3 mice (n=6/genotype) were examined for amyloid burden and neurofibrillary alterations, by using 6E10 immunohistochemistry and the Gallyas silver stain, respectively. Serotonin transporter (SERT) densities in the dorsal and the median raphe were quantified by [3H]DASB autoradiography. SERT mRNA expression was measured by RT-PCR and visualized by in situ hybridization. Neuroinflammation was evaluated by immunohistochemical staining for microglia and astrocytes, and by measuring mRNA levels of the proinflammatory cytokines TNF-α, IL-1β and IL-6. Results: No amyloid- and tau-associated lesions were observed in the midbrain raphe of 12 month old APPswe/PS1dE9 mice. SERT binding levels were reduced in transgenic animals compared to age-matched controls, and SERT mRNA levels were decreased by at least 50% from control values. Intense microglial, but not astrocytic immunoreactivity was observed in APPswe/PS1dE9 vs. wild-type mice. Levels of TNF-α mRNA were two-fold higher than control and correlated positively with SERT mRNA expression levels in transgenic animals. Conclusions: There was no amyloid accumulation and tau-associated pathology in the midbrain raphe of 12 month old APPswe/PS1dE9 mice. However, there was a local neuroinflammatory response with loss of serotonergic markers, which may partially account for some of the behavioral symptoms of AD.

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Reciprocal Regulatory Interaction between TRPV1 and Kinin B1 Receptor in a Rat Neuropathic Pain Model

International Journal of Molecular Sciences ◽

10.3390/ijms21030821 ◽

2020 ◽

Vol 21 (3) ◽

pp. 821 ◽

Cited By ~ 1

Author(s):

Veronica Cernit ◽

Jacques Sénécal ◽

Rahmeh Othman ◽

Réjean Couture

Keyword(s):

Spinal Cord ◽

Neuropathic Pain ◽

Mrna Expression ◽

Thermal Hyperalgesia ◽

Mrna Levels ◽

Regulatory Interaction ◽

B1 Receptor ◽

Tnf Α ◽

Kinin B1 Receptor ◽

Sensory Fibers

Kinins are mediators of pain and inflammation and evidence suggests that the inducible kinin B1 receptor (B1R) is involved in neuropathic pain (NP). This study investigates whether B1R and TRPV1 are colocalized on nociceptors and/or astrocytes to enable regulatory interaction either directly or through the cytokine pathway (IL-1β, TNF-α) in NP. Sprague Dawley rats were subjected to unilateral partial sciatic nerve ligation (PSNL) and treated from 14 to 21 days post-PSNL with antagonists of B1R (SSR240612, 10 mg·kg−1, i.p.) or TRPV1 (SB366791, 1 mg·kg−1, i.p.). The impact of these treatments was assessed on nociceptive behavior and mRNA expression of B1R, TRPV1, TNF-α, and IL-1β. Localization on primary sensory fibers, astrocytes, and microglia was determined by immunofluorescence in the lumbar spinal cord and dorsal root ganglion (DRG). Both antagonists suppressed PSNL-induced thermal hyperalgesia, but only SB366791 blunted mechanical and cold allodynia. SSR240612 reversed PSNL-induced enhanced protein and mRNA expression of B1R and TRPV1 mRNA levels in spinal cord while SB366791 further increased B1R mRNA/protein expression. B1R and TRPV1 were found in non-peptide sensory fibers and astrocytes, and colocalized in the spinal dorsal horn and DRG, notably with IL-1β on astrocytes. IL-1β mRNA further increased under B1R or TRPV1 antagonism. Data suggest that B1R and TRPV1 contribute to thermal hyperalgesia and play a distinctive role in allodynia associated with NP. Close interaction and reciprocal regulatory mechanism are suggested between B1R and TRPV1 on astrocytes and nociceptors in NP.

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Enhanced AP-1 and NF-κB activities and stability of interleukin 8 (IL-8) transcripts are implicated in IL-8 mRNA superinduction in lung epithelial H292 cells

Biochemical Journal ◽

10.1042/bj3300429 ◽

1998 ◽

Vol 330 (1) ◽

pp. 429-435 ◽

Cited By ~ 85

Author(s):

Thierry ROGER ◽

A. Theo OUT ◽

Naofumi MUKAIDA ◽

Kouji MATSUSHIMA ◽

M. Henk JANSEN ◽

...

Keyword(s):

Protein Synthesis ◽

Dna Binding ◽

Mrna Expression ◽

Gene Transcription ◽

Molecular Mechanisms ◽

Interleukin 8 ◽

Mrna Levels ◽

Tnf Α ◽

Lung Epithelial ◽

Inhibitor Of Protein Synthesis

Inhibition of protein synthesis may result in superinduction of short-lived transcripts and has been attributed variably to stabilization of transcripts and/or increased gene transcription. Little is known about the kinetics of these processes and relevant transcriptional elements have not been identified. In this study, we describe superinduction of interleukin 8 (IL-8) mRNA, an important inflammatory mediator, in lung epithelial-like H292 cells and identify the underlying molecular mechanisms and their kinetics. Cycloheximide (CHI, 10 μg/ml), an inhibitor of protein synthesis, maximally increased IL-8 mRNA levels 30-fold in H292 cells. Tumour necrosis factor α (TNF-α), which induced IL-8 mRNA 3-fold, synergized with CHI causing a 150-fold increase at 6 h. CHI early on increased the stability of IL-8 mRNA (from 40 min in cells cultured with medium to more than 4 h with CHI). CHI also increased transcription as shown by transfection with IL-8 promoter constructs. Truncated and mutated constructs identified NF-κB and AP-1 binding sites as primary cis-acting elements in IL-8 gene transcription and IL-8 mRNA superinduction. Electrophoretic mobility shift assays indicated that CHI increased NF-κB and prolonged AP-1 DNA-binding activities and that the synergism of TNF-α and CHI on IL-8 mRNA expression was paralleled by a further increase of AP-1 DNA-binding activity. This synergism was still noticed when 4 h elapsed between the addition of CHI and that of TNF-α. Taken together, our results indicate that CHI interferes with both post-transcriptional and transcriptional repressive mechanisms of IL-8 mRNA expression.

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Histamine enhances the production of human β-defensin-2 in human keratinocytes

AJP Cell Physiology ◽

10.1152/ajpcell.00293.2007 ◽

2007 ◽

Vol 293 (6) ◽

pp. C1916-C1923 ◽

Cited By ~ 25

Author(s):

Naoko Kanda ◽

Shinichi Watanabe

Keyword(s):

Mrna Expression ◽

Wound Repair ◽

Human Keratinocytes ◽

Serine Phosphorylation ◽

Epidermal Keratinocytes ◽

Mrna Levels ◽

Tnf Α ◽

Ifn Γ ◽

Microbial Defense

The anti-microbial peptide human β-defensin-2 (hBD-2), produced by epidermal keratinocytes, plays pivotal roles in anti-microbial defense, inflammatory dermatoses, and wound repair. hBD-2 induces histamine release from mast cells. We examined the in vitro effects of histamine on hBD-2 production in normal human keratinocytes. Histamine enhanced TNF-α- or IFN-γ-induced hBD-2 secretion and mRNA expression. Histamine alone enhanced transcriptional activities of NF-κB and activator protein-1 (AP-1) and potentiated TNF-α-induced NF-κB and AP-1 activities or IFN-γ-induced NF-κB and STAT1 activities. Antisense oligonucleotides against NF-κB components p50 and p65, AP-1 components c-Jun and c-Fos, or H1 antagonist pyrilamine suppressed hBD-2 production induced by histamine plus TNF-α or IFN-γ. Antisense oligonucleotide against STAT1 only suppressed hBD-2 production induced by histamine plus IFN-γ. Histamine induced serine phosphorylation of inhibitory NF-κBα (IκBα) alone or together with TNF-α or IFN-γ. Histamine induced c-Fos mRNA expression alone or together with TNF-α, whereas it did not further increase c-Jun mRNA levels enhanced by TNF-α. Histamine induced serine phosphorylation of STAT1 alone or together with IFN-γ, whereas it did not further enhance IFN-γ-induced tyrosine phosphorylation of STAT1. The histamine-induced serine phosphorylation of STAT1 was suppressed by MAPKK (MEK) inhibitor PD98059. These results suggest that histamine stimulates H1 receptor and potentiates TNF-α- or IFN-γ-induced hBD-2 production dependent on NF-κB, AP-1, or STAT1 in human keratinocytes. Histamine may potentiate anti-microbial defense, skin inflammation, and wound repair via the induction of hBD-2.

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The Expression of Toll-Like Receptor 4 mRNA in PBMCs Is Upregulated in Smokers and Decreases Upon Smoking Cessation

Frontiers in Immunology ◽

10.3389/fimmu.2021.667460 ◽

2021 ◽

Vol 12 ◽

Author(s):

Hsin-Yu Yeh ◽

Shou-Hung Hung ◽

Su-Chiu Chen ◽

Fei-Ran Guo ◽

Hsien-Liang Huang ◽

...

Keyword(s):

Smoking Cessation ◽

Cigarette Smoking ◽

Mrna Expression ◽

Mononuclear Cells ◽

Intercellular Adhesion Molecule ◽

Mrna Levels ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Tlr4 Mrna ◽

Tnf Α

BackgroundStudies have shown in vitro that cigarette smoke condensate stimulates monocytes to express toll-like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule 1 (ICAM-1), and enhances their adhesion to the endothelium. However, the same effects of cigarette smoking have not been explored in vivo. This study is to investigate the effect of cigarette smoking and smoking cessation on their mRNA expression in human peripheral blood mononuclear cells (PBMCs).MethodsA group of 97 smokers and 62 nonsmokers were enrolled. The RNA from PBMCs was assessed with real-time polymerase chain reaction (PCR) to determine the levels of ICAM-1, TNF-α, and TLR4. The same markers in PBMCs of 87 quitters were examined before and at one week, one month, and two months after smoking cessation.ResultsOf the 97 smokers, 85 (87.6%) were males, and 30 (48.4%) of the nonsmokers were males (p < 0.0001). The mean (SD) age of the smokers was 43.24 (10.89) years, which was younger than 43.45 (11.41) years of nonsmokers (p < 0.0001). The incidence of cardiovascular diseases was 13.4% in smokers, which was higher than 1.6% in nonsmokers (p < 0.05). Both ICAM-1 and TNF-α mRNA levels in PBMCs were higher among the smokers (p < 0.0001). In addition, TLR4 mRNA levels in PBMCs were statistically elevated in the smokers (p < 0.0001) comparing with those in the nonsmokers. The mRNA levels of TLR4 and TNF-α in PBMCs decreased in those who had quit smoking for 2 months (p < 0.0001).ConclusionsICAM-1, TNF-α, and TLR4 mRNA expression levels in PBMCs increased in smokers and decreased after being on a smoking cessation program for 2 months. This finding suggested that TLR4 expression may mediate the atherogenic inflammatory process induced by smoking.

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Brief report: Expression of TNF-α in chronic periapical lesions correlates with expression of bacterial chaperonin-60

Vojnosanitetski pregled ◽

10.2298/vsp201106052s ◽

2021 ◽

pp. 52-52

Author(s):

Jelena Stanisic-Zindovic ◽

Branko Mihailovic ◽

Filip Djordjevic ◽

Marija Milovanovic ◽

Nebojsa Arsenijevic ◽

...

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Mrna Expression ◽

Mrna Levels ◽

Periapical Lesions ◽

Quantitative Expression ◽

Tnf Α ◽

Chaperonin 60 ◽

Anti Inflammatory ◽

Anti Inflammatory Cytokine

Background/Aim: The aim of this study is to determine the quantitative expression of the bacterial heat shock protein, Chaperonin-60 (Cpn60) and pro-inflammatory and anti-inflammatory cytokine in periapical tissue, obtained from individuals with chronic periapical lesions and to determine the correlation between the expression of the bacterial heat shock protein and the expression of these cytokines. Methods. The study was performed on 18 periapical lesions and 6 control samples of healthy periapical tissue, taken at the Clinic of Dental Medicine, Faculty of Medical 4 Sciences University of Pristina, Kosovska Mitrovica. The levels of mRNA expression of pro- and anti- inflammatory cytokines and bacterial heat shock protein were determined by real time quantitative RT-PCR. Results. Analysis revealed significantly higher mRNA levels of TNF-? and Cpn60 in the tissue of periapical lesions compared with normal periapical tissue (P <0.05). Contrary to these results, the mRNA expression of anti-inflammatory IL-10 was significantly higher in the samples of normal periapical tissue compared with the mRNA levels of this cytokine in the tissue of periapical lesions (P <0.001). Expression of Cpn60 is in strong correlation with TNF-? expression in periapical lesions. Conclusion. Cpn60 released from bacteria in periapical tissue could be a strong stimulator of inflammatory response and one of the important players in the pathogenesis of periapical lesions.

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Intra-uterine Growth Restriction Downregulates the Hepatic Toll Like Receptor-4 Expression and Function

Clinical and Developmental Immunology ◽

10.1080/17402520400008905 ◽

2005 ◽

Vol 12 (1) ◽

pp. 59-66 ◽

Cited By ~ 14

Author(s):

Ozlem Equils ◽

Sapna Singh ◽

Semra Karaburun ◽

Daning Lu ◽

Manikkavasagar Thamotharan ◽

...

Keyword(s):

Mrna Expression ◽

Growth Restriction ◽

Milk Intake ◽

Mrna Levels ◽

Toll Like Receptor ◽

Tlr4 Expression ◽

Tlr4 Mrna ◽

Increased Risk ◽

Tnf Α ◽

And Function

Maternal starvation is a significant cause of intrauterine growth restriction (IUGR) in the world and increases the risk of infection in the neonate. We examined the effect of maternal starvation on Toll like receptor (TLR)4 expression in hepatic, splenic and intestinal tissues obtained from the adult IUGR offspring of prenatal calorie restricted rats. The hepatic TLR4 protein concentration was undetectable in the IUGR rats that had restricted milk intake during the suckling period (SM/SP;n= 4,p< 0.05) as compared to the normal growth controls (CM/CP;n=4), and access to ad lib milk intake during the sucking period partially corrected the hepatic TLR4 expression (SM/CP;n= 4). IUGR had no effect on the splenic (n= 4) or intestinal (n= 4) TLR4 mRNA levels. In the liver, IUGR led to a 20% increase in baseline tumor necrosis factor (TNF)-α mRNA expression (p< 0.03) and a 70% increase in interleukin-1β (IL-1β) mRNA expression (p< 0.008) as compared to the control rats (CM/CP;n= 7). LPS-induced hepatic TNF-α release was significantly higher in SM/SP as compared to CM/CP. We propose that IUGR dysregulates TLR4 expression and function in the offspring, which may help explain the increased risk of Gram-negative sepsis and inflammatory diseases in this population.

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Cytokine Expression in the Colonic Mucosa of Human Immunodeficiency Virus-Infected Individuals before and during 9 Months of Antiretroviral Therapy

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00250-08 ◽

2008 ◽

Vol 52 (9) ◽

pp. 3377-3384 ◽

Cited By ~ 23

Author(s):

Hubert Schulbin ◽

Hagen Bode ◽

Hartmut Stocker ◽

Wolfgang Schmidt ◽

Thomas Zippel ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Antiretroviral Therapy ◽

T Lymphocytes ◽

Mrna Expression ◽

Colonic Mucosa ◽

Mrna Levels ◽

Ifn Gamma ◽

Hiv Rna ◽

Tnf Α ◽

Immunodeficiency Virus

ABSTRACT High-level human immunodeficiency virus (HIV) replication and the rapid breakdown of the mucosal immune system are the hallmarks of HIV infection in the gut. Cytokine dysregulation may be related to both phenomena. Using real-time PCR we quantified the colonic mucosal mRNA expression of selected proinflammatory and regulatory (gamma interferon [IFN-gamma], tumor necrosis factor alpha [TNF-α], and interleukin-2 [IL-2], IL-4, IL-6, and IL-10) and HIV-inhibitory (IL-16, CCL3, and CCL5) cytokines for 10 HIV-infected patients before and during 9 months of highly active antiretroviral therapy (HAART). HIV RNA and T-cell dynamics were measured in the colonic mucosa and the blood. Seven HIV-negative individuals served as controls. The mucosal mRNA expression of TNF-α, IFN-gamma, IL-4, IL-6, and IL-10 was significantly higher in HIV-infected patients than in control patients and remained elevated during 9 months of HAART despite the decline in blood and mucosal HIV RNA levels and an increase in the level of CD4+ T lymphocytes. The mRNA levels of CCL3 and CCL5, both of which were elevated before treatment, returned to nearly normal during therapy. Despite reductions in levels of mucosal HIV RNA and the restoration of mucosal CD4+ T lymphocytes, antiretroviral therapy failed to restore the normal colonic immunologic environment.

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Cytokine-dependent regulation of hepatic organic anion transporter gene transactivators in mouse liver

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00307.2004 ◽

2005 ◽

Vol 289 (5) ◽

pp. G831-G841 ◽

Cited By ~ 70

Author(s):

Andreas Geier ◽

Christoph G. Dietrich ◽

Sebastian Voigt ◽

Meenakshisundaram Ananthanarayanan ◽

Frank Lammert ◽

...

Keyword(s):

Mrna Expression ◽

Hormone Receptors ◽

Organic Anion ◽

Pregnane X Receptor ◽

Regulatory Elements ◽

Farnesoid X Receptor ◽

Organic Anion Transporter ◽

Mrna Levels ◽

Anion Transporter ◽

Tnf Α

Proinflammatory cytokines such as TNF-α and IL-1β lead to downregulation of hepatic organic anion transporters in cholestasis. This adapted response is transcriptionally mediated by nuclear hormone receptors and liver-specific transcription factors. Because little is known in vivo about cytokine-dependent regulatory events, mice were treated with either TNF-α or IL-1β for up to 16 h. Transporter mRNA expression was determined by Northern blot analysis, nuclear activity, and protein-expression of transactivators by EMSA and Western blotting. TNF-α induces a sustained decrease in Ntcp, Oatp1/Oatp1a1, and Bsep mRNA expression but exerts only transient [multidrug resistance-associated protein 2 (Mrp2)] or no effects (Mrp3) on Mrps. In addition to Ntcp and Oatp1/Oatp1a1, IL-1β also downregulates Bsep, Mrp2, and Mrp3 mRNAs to some extent. To study transcriptional regulation, Ntcp and Bsep promoters were first cloned from mice revealing a new distal Ntcp hepatocyte nuclear factor 1 (HNF-1) element but otherwise show a conserved localization to known rat regulatory elements. Changes in transporter-expression are preceeded by a reduction in binding activities at IR-1, ER-8, DR-5, and HNF-1α sites after 4 h by either cytokine, which remained more sustained by TNF-α in the case of nuclear receptors. Nuclear protein levels of retinoid X receptor (RXR)-α are significantly decreased by TNF-α but only transiently affected by IL-1β. Minor reductions of retinoic acid receptor, farnesoid X receptor, pregnane X receptor, and constitutive androstane receptor nuclear proteins are restricted to 4 h after cytokine application and paralleled by a decrease in mRNA levels. Basolateral and canalicular transporter systems are downregulated by both cytokines, TNF-α and IL-1β. Activity of HNF-1α as regulator of mNtcp is suppressed by both cytokines. Decreased binding activities of nuclear receptor heterodimers may be explained by a reduction of the ubiquitous heterodimerization partner RXR-α.

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Interleukin-1α stimulates proinflammatory cytokine expression in human cardiac myofibroblasts

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00372.2009 ◽

2009 ◽

Vol 297 (3) ◽

pp. H1117-H1127 ◽

Cited By ~ 69

Author(s):

Neil A. Turner ◽

Anupam Das ◽

Philip Warburton ◽

David J. O'Regan ◽

Stephen G. Ball ◽

...

Keyword(s):

Protein Kinase ◽

Mrna Expression ◽

Signaling Pathways ◽

P38 Mapk ◽

Proinflammatory Cytokines ◽

Enzyme Linked Immunosorbent Assay ◽

Mrna Levels ◽

Tnf Α ◽

Interleukin 1Α ◽

Cardiac Myofibroblasts

Cardiac myofibroblasts (CMF) play a key role in infarct repair and scar formation following myocardial infarction (MI) and are also an important source of proinflammatory cytokines. We postulated that interleukin-1α (IL-1α), a potential early trigger of acute inflammation post-MI, could stimulate human CMF to express additional proinflammatory cytokines. Furthermore, we hypothesized that these effects may be modulated by the anti-inflammatory cytokine interleukin-10 (IL-10). Human CMF were cultured from atrial biopsies from multiple patients. Interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and cardiotrophin-1 (CT-1) mRNA expression and secretion were measured using quantitative real-time RT-PCR and enzyme-linked immunosorbent assay. IL-1α (0.001–10 ng/ml, 0–6 h) stimulated IL-1β, TNF-α, and IL-6 mRNA expression with distinct temporal and concentration profiles, resulting in increased cytokine secretion. The response to IL-1α was much greater than with TNF-α. Neither IL-1α nor TNF-α treatment modulated CT-1 mRNA expression. Immunoblotting with phosphospecific antibodies revealed that IL-1α stimulated the extracellular signal-regulated kinase (ERK)-1/2, p38 mitogen-activated protein kinase (MAPK), c-Jun NH2-terminal kinase (JNK), phosphatidylinositol 3-kinase (PI 3-kinase)/protein kinase B (Akt), and nuclear factor (NF)-κB signaling pathways. Pharmacological inhibitor studies indicated roles for PI 3-kinase/Akt and NF-κB pathways in mediating IL-1β expression, and for NF-κB and p38 MAPK pathways in mediating TNF-α expression. IL-1α-induced IL-6 mRNA expression was reduced by p38 MAPK inhibition, but increased by ERK and JNK pathway inhibitors. IL-10 produced a consistent but modest reduction in IL-1α-induced IL-6 mRNA levels (not IL-1β or TNF-α), but this was not reflected by reduced IL-6 protein secretion. In conclusion, IL-1α stimulates human CMF to express IL-1β, TNF-α, and IL-6 via specific signaling pathways, responses that are unaffected by IL-10 exposure.

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