scholarly journals Pre- and post-translational negative effect of β-adrenoceptor agonists on adiponectin secretion: in vitro and in vivo studies

2002 ◽  
Vol 367 (3) ◽  
pp. 677-685 ◽  
Author(s):  
Marie-Laure DELPORTE ◽  
Tohru FUNAHASHI ◽  
Masahiko TAKAHASHI ◽  
Yuji MATSUZAWA ◽  
Sonia M. BRICHARD
Keyword(s):  
Gene Expression ◽  
In Vivo Studies ◽  
Adrenergic Agonists ◽  
Cytosol Fraction ◽  
Negative Effect ◽  
Adrenoceptor Agonists ◽  
Visceral Adipose ◽  
Qualitative Changes

The adipose-derived hormone, adiponectin (ApN), has a role in fuel homoeostasis, insulin action and atherosclerosis. Regulation of ApN by catecholamines has scarcely been investigated. We examined the effects of β-adrenergic agonists (and their second messenger, cAMP) on ApN gene expression, production and secretion in mouse in vitro and in vivo; their effects in human fat were also briefly studied in vitro. β-Adrenergic agonists and cAMP inhibited ApN gene expression in human visceral adipose tissue. Likewise, cAMP down-regulated ApN mRNAs in cultured mouse explants from visceral and subcutaneous regions. The amount of ApN released into the medium decreased concomitantly. cAMP also caused qualitative changes in ApN secretion. Under basal conditions, ApN was secreted as a single 32kDa species. In the presence of cAMP, an additional and probably immature (not modified post-translationally) 30kDa species was also sorted. This altered secretion resulted from cAMP-induced quantitative and qualitative changes of ApN within the adipocyte. Under basal conditions, the 32kDa form of ApN was mainly associated with high-density microsomes (HDMs), while the 30kDa species was confined to a pool recovered with the cytosol fraction. cAMP depleted intracellular ApN at the expense of both HDM and cytosol fractions, and abnormally targeted ApN species to the different subcellular compartments as a result of impaired maturation. β-Adrenergic agonists mimicked the inhibitory effects of cAMP on ApN mRNA and secretion, the β3-agonist BRL37344 being the most potent. Administration of BRL37344 to mice reduced ApN mRNAs in both adipose regions, and ApN levels in plasma. In conclusion, β-agonists inhibited ApN production and maturation, and thus exerted a dual (pre- and post-translational) negative effect on ApN secretion by cultured mouse adipose explants. ApN inhibition by β-agonists was reproduced in mouse in vivo and in humans in vitro. ApN down-regulation may have an important role in fuel homoeostasis, insulin resistance and stress-induced atherosclerosis.

scholarly journals CircFAM13B promotes the proliferation of hepatocellular carcinoma by sponging miR-212, upregulating E2F5 expression and activating the P53 pathway

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ying Xie ◽  
Xiaofeng Hang ◽  
Wensheng Xu ◽  
Jing Gu ◽  
Yuanjing Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepatocellular Carcinoma ◽  
Gene Expression Omnibus ◽  
Differentially Expressed ◽  
In Vivo Studies ◽  
Circular Rnas ◽  
Novel Target ◽  
Underlying Mechanisms

Abstract Background Most of the biological functions of circular RNAs (circRNAs) and the potential underlying mechanisms in hepatocellular carcinoma (HCC) have not yet been discovered. Methods In this study, using circRNA expression data from HCC tumor tissues and adjacent tissues from the Gene Expression Omnibus database, we identified out differentially expressed circRNAs and verified them by qRT-PCT. Functional experiments were performed to evaluate the effects of circFAM13B in HCC in vitro and in vivo. Results We found that circFAM13B was the most significantly differentially expressed circRNA in HCC tissue. Subsequently, in vitro and in vivo studies also demonstrated that circFAM13B promoted the proliferation of HCC. Further studies revealed that circFAM13B, a sponge of miR-212, is involved in the regulation of E2F5 gene expression by competitively binding to miR-212, inhibits the activation of the P53 signalling pathway, and promotes the proliferation of HCC cells. Conclusions Our findings revealed the mechanism underlying the regulatory role played by circFAM13B, miR-212 and E2F5 in HCC. This study provides a new theoretical basis and novel target for the clinical prevention and treatment of HCC.

scholarly journals Modulatory Effects of Osthole on Lipopolysaccharides-Induced Inflammation in Caco-2 Cell Monolayer and Co-Cultures with THP-1 and THP-1-Derived Macrophages

Nutrients ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 123
Author(s):  
Natalia K. Kordulewska ◽  
Justyna Topa ◽  
Małgorzata Tańska ◽  
Anna Cieślińska ◽  
Ewa Fiedorowicz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Inflammatory Reaction ◽  
Wide Spectrum ◽  
Cell Monolayer ◽  
In Vivo Studies ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Tnf Α

Lipopolysaccharydes (LPS) are responsible for the intestinal inflammatory reaction, as they may disrupt tight junctions and induce cytokines (CKs) secretion. Osthole has a wide spectrum of pharmacological effects, thus its anti-inflammatory potential in the LPS-treated Caco-2 cell line as well as in Caco-2/THP-1 and Caco-2/macrophages co-cultures was investigated. In brief, Caco-2 cells and co-cultures were incubated with LPS to induce an inflammatory reaction, after which osthole (150–450 ng/mL) was applied to reduce this effect. After 24 h, the level of secreted CKs and changes in gene expression were examined. LPS significantly increased the levels of IL-1β, -6, -8, and TNF-α, while osthole reduced this effect in a concentration-dependent manner, with the most significant decrease when a 450 ng/mL dose was applied (p < 0.0001). A similar trend was observed in changes in gene expression, with the significant osthole efficiency at a concentration of 450 ng/μL for IL1R1 and COX-2 (p < 0.01) and 300 ng/μL for NF-κB (p < 0.001). Osthole increased Caco-2 monolayer permeability, thus if it would ever be considered as a potential drug for minimizing intestinal inflammatory symptoms, its safety should be confirmed in extended in vitro and in vivo studies.

IWR-1 Inhibits Collagen-Induced Platelet Activation and Protects against Thrombogenesis

2019 ◽  
Vol 39 (04) ◽  
pp. 392-397
Author(s):  
Wei Wang ◽  
Songqing Lai ◽  
ZiJin Xiao ◽  
Haiyue Yan ◽  
Yongxi Li ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Antiplatelet Agent ◽  
Wnt Signalling ◽  
In Vivo Studies ◽  
Dependent Manner ◽  
Platelet Activity ◽  
Occlusion Time ◽  
Negative Effect

AbstractPlatelets play a crucial role in haemostasis and several pathophysiological processes. Collagen is a main initiator for platelet activation and aggregation. Given that Wnt signalling negatively regulates platelet function, and IWR-1 (a small molecule inhibitor for Wnt signalling) has the potential of inhibiting collagen synthesis, it is essential to investigate whether IWR-1 regulates collagen-induced platelet activation and protects against thrombogenesis. In the present study we found that IWR-1 pretreatment effectively suppressed collagen-induced platelet aggregation in a dose-dependent manner. In addition, IWR-1 also resulted in a decrease of P-selectin and phosphatidylserine surface exposure using fluorescence-activated cell sorting analysis. In vitro studies further revealed that IWR-1 had a negative effect on integrin a2β1 activation and platelet spreading. More importantly, the results from in vivo studies showed that IWR-1 exhibited a robust bleeding diathesis in the tail-bleeding assay and a prolonged occlusion time in the FeCl3-induced carotid injury model. Taken together, current results demonstrate that IWR-1 could effectively block collagen-induced platelet activity in vitro and in vivo, and suggest its candidacy as a new antiplatelet agent.

The VSL#3 probiotic formula induces mucin gene expression and secretion in colonic epithelial cells

2007 ◽  
Vol 292 (1) ◽  
pp. G315-G322 ◽  
Author(s):  
C. Caballero-Franco ◽  
K. Keller ◽  
C. De Simone ◽  
K. Chadee
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Cultured Cells ◽  
In Vivo Studies ◽  
Lactobacillus Species ◽  
Mucin Secretion ◽  
Colonic Epithelial Cells ◽  
Dnase Treatment

Several studies have stressed the importance of the microbiota in the maintenance of the gastrointestinal epithelium. Administration of probiotic bacteria, supplements composed of microbiota constituents, was previously shown to diminish symptoms in patients suffering from inflammatory bowel diseases. This raises the possibility that probiotics may play an active role in enhancing the intestinal barrier at the mucosal surface. In this study, we investigated whether the clinically tested VSL#3 probiotic formula and/or its secreted components can augment the protective mucus layer in vivo and in vitro. For in vivo studies, Wistar rats were orally administered the probiotic mixture VSL#3 on a daily basis for seven days. After treatment, basal luminal mucin content increased by 60%. In addition, we exposed isolated rat colonic loops to the VSL#3 probiotic formula, which significantly stimulated colonic mucin (MUC) secretion and MUC2 gene expression; however, MUC1 and MUC3 gene expression were only slightly elevated. The effect of the VSL#3 mucin secretagogue was also tested in vitro by use of LS 174T colonic epithelial cells. In contrast to the animal studies, cultured cells incubated with VSL#3 bacteria did not exhibit increased mucin secretion. However, the bacterial secreted products contained in the conditioned media stimulated a remarkable mucin secretion effect. Among the three bacterial groups ( Lactobacilli, Bifidobacteria, and Streptococci) contained in VSL#3, the Lactobacillus species were the strongest potentiator of mucin secretion in vitro. A preliminary characterization of the putative mucin secretagogue suggested that it was a heat-resistant soluble compound, which is not sensitive to protease and DNase treatment. These findings contribute to a better understanding of the complex and beneficial interaction between colonic epithelial cells and intestinal bacteria.

Antisense Oligodeoxynucleotide Technology: Potential Use for the Treatment of Malignant Brain Tumors

1998 ◽  
Vol 5 (2) ◽  
pp. 163-170 ◽  
Author(s):  
Herbert H. Engelhard
Keyword(s):  
Gene Expression ◽  
Brain Tumors ◽  
Relevant Information ◽  
In Vivo Studies ◽  
Antisense Oligodeoxynucleotide ◽  
Malignant Brain Tumors ◽  
Antisense Odns ◽  
The Brain

Background: Antisense oligodeoxynucleotides (ODNs) have been proposed as a new therapy for patients with cancer, including malignant brain tumors. Antisense ODNs are taken up by tumor cells and selectively block gene expression. Use of ODNs for brain tumors is attractive due to their theoretical specificity, relative ease of production and, to date, paucity of reported adverse effects. This article presents current information regarding antisense ODNs and their possible future use for the treatment of brain tumors. Methods: The available published experimental and clinical information regarding antisense ODN treatment of glioblastoma cells and administration into the central nervous system (CNS) was reviewed. Other clinically relevant information pertaining to the molecular biology of antisense ODNs was also collected and summarized. Results: Targets for antisense ODN therapy in malignant glioma cells have included c-myc, c-myb, c-sis, c-erb B, CD44, p34cdc2, bFGF, PDGF, TGF-beta, IGF-1, PKC-alpha tumor necrosis factor, urokinase, and S100beta protein. Few in vivo studies of ODN treatment of brain tumors have yet been reported. Systemically administered ODNs enter the brain only in extremely small quantities; therefore, microinfusion into the brain has been recommended. Conclusions: Antisense ODNs have been used successfully to block glioblastoma gene expression in vitro and expression of multiple genes within the CNS of experimental animals. Upcoming clinical trials will address the safety of antisense ODN use against malignant brain tumors.

scholarly journals Yeast TFIID Serves as a Coactivator for Rap1p by Direct Protein-Protein Interaction

2006 ◽  
Vol 27 (1) ◽  
pp. 297-311 ◽  
Author(s):  
Krassimira A. Garbett ◽  
Manish K. Tripathi ◽  
Belgin Cencki ◽  
Justin H. Layer ◽  
P. Anthony Weil
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Gene Transcription ◽  
In Vitro Transcription ◽  
Tata Binding Protein ◽  
In Vivo Studies ◽  
Activator Protein 1 ◽  
Protein Protein Interaction

ABSTRACT In vivo studies have previously shown that Saccharomyces cerevisiae ribosomal protein (RP) gene expression is controlled by the transcription factor repressor activator protein 1 (Rap1p) in a TFIID-dependent fashion. Here we have tested the hypothesis that yeast TFIID serves as a coactivator for RP gene transcription by directly interacting with Rap1p. We have found that purified recombinant Rap1p specifically interacts with purified TFIID in pull-down assays, and we have mapped the domains of Rap1p and subunits of TFIID responsible. In vitro transcription of a UASRAP1 enhancer-driven reporter gene requires both Rap1p and TFIID and is independent of the Fhl1p-Ifh1p coregulator. UASRAP1 enhancer-driven transactivation in extracts depleted of both Rap1p and TFIID is efficiently rescued by addition of physiological amounts of these two purified factors but not TATA-binding protein. We conclude that Rap1p and TFIID directly interact and that this interaction contributes importantly to RP gene transcription.

scholarly journals Bile acids inhibit duodenal secretin expression via orphan nuclear receptor small heterodimer partner (SHP)

2009 ◽  
Vol 297 (1) ◽  
pp. G90-G97 ◽  
Author(s):  
Ian P. Y. Lam ◽  
Leo T. O. Lee ◽  
Hueng-Sik Choi ◽  
Gianfranco Alpini ◽  
Billy K. C. Chow
Keyword(s):  
Gene Expression ◽  
Bile Acids ◽  
Nuclear Receptor ◽  
Target Genes ◽  
In Vivo Studies ◽  
Control Group ◽  
Orphan Nuclear Receptor ◽  
Small Heterodimer Partner

Small heterodimer partner (SHP) is an orphan nuclear receptor in which gene expression can be upregulated by bile acids. It regulates its target genes by repressing the transcriptional activities of other nuclear receptors including NeuroD, which has been shown to regulate secretin gene expression. Here, we evaluated the regulation on duodenal secretin gene expression by SHP and selected bile acids, cholic acid (CA) and chenodeoxycholic acid (CDCA). In vitro treatment of CDCA or fexaramine elevated the SHP transcript level and occupancy on secretin promoter. The increase in the SHP level, induced by bile acid treatment or overexpression, reduced secretin gene expression, whereas this gene inhibitory effect was reversed by silencing of endogenous SHP. In in vivo studies, double-immunofluorescence staining demonstrated the coexpression of secretin and SHP in mouse duodenum. Feeding mice with 1% CA-enriched rodent chow resulted in upregulation of SHP and a concomitant decrease in secretin transcript and protein levels in duodenum compared with the control group fed with normal chow. A diet enriched with 5% cholestyramine led to a decrease in SHP level and a corresponding increase in secretin expression. Overall, this study showed that bile acids via SHP inhibit duodenal secretin gene expression. Because secretin is a key hormone that stimulates bile flow in cholangiocytes, this pathway thus provides a novel means to modulate secretin-stimulated choleresis in response to intraduodenal bile acids.

scholarly journals EDR Peptide: Possible Mechanism of Gene Expression and Protein Synthesis Regulation Involved in the Pathogenesis of Alzheimer’s Disease

Molecules ◽  
2020 ◽  
Vol 26 (1) ◽  
pp. 159
Author(s):  
Vladimir Khavinson ◽  
Natalia Linkova ◽  
Ekaterina Kozhevnikova ◽  
Svetlana Trofimova
Keyword(s):  
Gene Expression ◽  
Signaling Pathway ◽  
Animal Studies ◽  
Enzyme Synthesis ◽  
In Vivo Studies ◽  
Neuronal Cultures ◽  
Ribonucleic Acids ◽  
Synthesis Regulation

The EDR peptide (Glu-Asp-Arg) has been previously established to possess neuroprotective properties. It activates gene expression and synthesis of proteins, involved in maintaining the neuronal functional activity, and reduces the intensity of their apoptosis in in vitro and in vivo studies. The EDR peptide interferes with the elimination of dendritic spines in neuronal cultures obtained from mice with Alzheimer’s (AD) and Huntington’s diseases. The tripeptide promotes the activation of the antioxidant enzyme synthesis in the culture of cerebellum neurons in rats. The EDR peptide normalizes behavioral responses in animal studies and improves memory issues in elderly patients. The purpose of this review is to analyze the molecular and genetics aspects of the EDR peptide effect on gene expression and synthesis of proteins involved in the pathogenesis of AD. The EDR peptide is assumed to enter cells and bind to histone proteins and/or ribonucleic acids. Thus, the EDR peptide can change the activity of the MAPK/ERK signaling pathway, the synthesis of proapoptotic proteins (caspase-3, p53), proteins of the antioxidant system (SOD2, GPX1), transcription factors PPARA, PPARG, serotonin, calmodulin. The abovementioned signaling pathway and proteins are the components of pathogenesis in AD. The EDR peptide can be AD.

scholarly journals CSIG-16. RECONCILING TUMOR HETEROGENEITY IN GLIOBLASTOMA USING A PATHWAY-BASED APPROACH

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi47-vi47
Author(s):  
Alvaro Alvarado ◽  
Riki Kawaguchi ◽  
Giovanni Coppola ◽  
Steven Goldman ◽  
Harley Kornblum
Keyword(s):  
Gene Expression ◽  
Gene List ◽  
Molecular Targets ◽  
Principal Component ◽  
Gene Set Enrichment Analysis ◽  
The Cancer Genome Atlas ◽  
In Vivo Studies ◽  
Molecular Architecture

Abstract Despite efforts to gain a deeper understanding of its molecular architecture, glioblastoma (GBM) remains uniformly fatal. While genome-based molecular subtyping has revealed that GBMs may be parsed into several distinct molecular categories, this insight has yielded little progress towards extending patient survival. In particular, the great phenotypic heterogeneity of GBM – both inter and intratumorally – has hindered therapeutic efforts. To this end, we interrogated tumor samples using a pathway-based approach to resolve tumoral heterogeneity. Gene set enrichment analysis (GSEA) was applied to gene expression data and used to provide an overview of each sample that was then compared to others, generating sample clusters based on overall patterns of enrichment. The Cancer Genome Atlas (TCGA) samples were clustered using canonical and oncogenic signatures and in both cases the clustering was distinct from the molecular subtypes previously reported. Using principal component analysis (PCA) and other bioinformatics tools, we extracted gene sets to further characterize the pathways contributing to each of these clusters. We generated gene lists of the top common elements and Ingenuity pathway analysis exposed molecular targets that control critical pathways of each identified cluster. Similar analyses were completed in a gene expression database of patient-derived gliomasphere lines and molecular targets were also obtained. We found E2F1 to be a strong target based on gene lists from both databases. A cluster of gliomasphere lines have high enrichment scores for the gene list predicted to depend on E2F1. In vitro genetic perturbation showed decrease stem cell frequency and lower expression of cell cycle progression genes in cell lines from this cluster exclusively. Other cluster-specific targets are being validated and in vivo studies will follow momentarily. Our studies relate intertumoral heterogeneity to critical cellular pathways dysregulated in GBM, with the ultimate goal of establishing a pipeline for patient- and tumor-specific precision medicine.

scholarly journals Plasticity of Proinflammatory Macrophages Depends on Their Polarization Stage during Human MSC Immunomodulation—An In Vitro Study Using THP-1 and Human Primary Macrophages

Immuno ◽  
2021 ◽  
Vol 1 (4) ◽  
pp. 518-528
Author(s):  
Constantin Ißleib ◽  
Susanne Kurz ◽  
Samuel Scholl ◽  
Bettina Amberg ◽  
Juliane Spohn
Keyword(s):  
Gene Expression ◽  
Human Mesenchymal Stem Cells ◽  
In Vitro Study ◽  
Cytokine Secretion ◽  
In Vivo Studies ◽  
M1 Macrophages ◽  
M1 Polarization ◽  
In Vitro Investigation

Human mesenchymal stem cells (hMSCs) are well-known for their immunomodulatory potential. In recent clinical trials and in vivo studies, hMSCs were used as therapeutic measures to dampen inflammation. In this context, their effect on macrophages in vivo has been described to induce a phenotype change shifting from a proinflammatory to an anti-inflammatory environment. Despite several in vitro studies that investigated the potential of hMSCs to inhibit the polarization of macrophages into the proinflammatory M1 subtype, it is still unclear if hMSCs affect polarized M1 macrophages or if they control the environment by inhibiting the M1 polarization of unpolarized macrophages. Here, a comparative in vitro investigation of hMSC immunomodulation via soluble factors concerning the influence on the polarization of macrophages to M1 and on polarized M1 macrophages is presented. Human primary monocyte-derived macrophages (hMDMs) as well as THP-1 cells were used for this investigation. The macrophage subtype was analyzed by gene expression as well as cytokine secretion. hMSCs affected cytokine secretion of polarizing macrophages, while changes in gene expression were evident in polarized M1 macrophages. These effects were observed in THP-1 and hMDM macrophages. In conclusion, we suggest that hMSCs implement their immunomodulatory effects on polarizing and polarized macrophages in different ways.

Sign in / Sign up

Export Citation Format

Share Document