scholarly journals The expression of cytokines in the milk somatic cells, blood leukocytes and serum of goats infected with small ruminant lentivirus

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Justyna Jarczak ◽  
Danuta Słoniewska ◽  
Jarosław Kaba ◽  
Emilia Bagnicka
Keyword(s):  
Protein Expression ◽  
Small Ruminant ◽  
Somatic Cells ◽  
Immunological Response ◽  
Dairy Goats ◽  
Blood Leukocytes ◽  
Protein Levels ◽  
Tnf Α ◽  
Gene And Protein Expression ◽  
Milk Somatic Cells

Abstract Background The present study aimed to determine the expression of cytokines, which is associated with the immunological response of dairy goats against small ruminant lentivirus (SRLV). The study was conducted on 26 dairy goats in their second to sixth lactation, which were divided by breed and parity into two groups: SRLV naturally infected (N = 13) and non-infected (N = 13) animals. All goats in the study were asymptomatic. The milk and blood samples, which served as studied material were taken on days 7, 30, 120 and 240 of the lactation. The gene and protein expression of several cytokines was studied using Real-Time PCR and ELISA methods. Results INF-β and INF-γ expression was down-regulated in the milk somatic cells (MSC) of SRLV-infected goats. However, an increased concentration of INF-β was observed in the MSC in SRLV-infected goats, while INF-γ expression was not observed in both SRLV-infected and non-infected animals The SRLV-infected goats also displayed decreased expression of IL-1α, IL-1β, IL-6 and INF-γ genes in the blood leukocytes,with IL-1α, IL-1β and IL-6 protein levels also being decreased in the sera. TNF-α was the only gene that demonstrated increased expression in both the MSC and the blood of infected animals; however, no such overexpression was observed at the protein level. Conclusions SRLV probably influences the immune system of infected animals by deregulating of the expression of cytokines. Further, epigenetic studies may clarify the mechanisms by which SRLV regulates the gene and protein expression of the host.

scholarly journals Crocin ameliorated skin tissue inflammation in atopic dermatitis in mice

2020 ◽  
Vol 6 (2) ◽  
pp. 142
Author(s):  
Abdullah Alyoussef
Keyword(s):  
Atopic Dermatitis ◽  
Protein Expression ◽  
Skin Disease ◽  
Therapeutic Effects ◽  
Inflammatory Skin Disease ◽  
Mice Model ◽  
Subsequent Reduction ◽  
Gardenia Jasminoides ◽  
Tnf Α ◽  
Gene And Protein Expression

<p class="abstract"><strong>Background:</strong> Atopic dermatitis (AD) is considered a chronic recurrent inflammatory skin disease. In addition, crocin is the major carotenoid compound found in Gardenia jasminoides. It is previously proved to produce anti-inflammatory actions. Therefore, we conducted this research to investigate the therapeutic effects of crocin on a mice model of AD.</p><p class="abstract"><strong>Methods:</strong> Mice were investigated for the number of scratches and dermatitis score. Skin was isolated and used for measurements of gene and protein expression of β-catenin, NFκB, TNF-α and IL-1β.<strong></strong></p><p class="abstract"><strong>Results:</strong> Authors found that crocin significantly reduced the number of scratches, ear thickness and dermatitis score. In addition, crocin ameliorated AD-induced elevation in the expression of β-catenin, NFκB, TNF-α and IL-1β.</p><p class="abstract"><strong>Conclusions:</strong> Crocin ameliorated DNCB-induced AD in mice via blockage of β-catenin with subsequent reduction in inflammatory pathway.</p><p class="abstract"> </p>

scholarly journals Silibinin Downregulates the NF-κB Pathway and NLRP1/NLRP3 Inflammasomes in Monocytes from Pregnant Women with Preeclampsia

Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1548 ◽  
Author(s):  
Mariana Leticia Matias ◽  
Virginia Juliani Gomes ◽  
Mariana Romao-Veiga ◽  
Vanessa Rocha Ribeiro ◽  
Priscila Rezeck Nunes ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Immunological Response ◽  
Response Characteristic ◽  
Sterile Inflammation ◽  
Blood Monocytes ◽  
Specific Syndrome ◽  
Tnf Α ◽  
Gene And Protein Expression ◽  
Nlrp3 Inflammasomes

Preeclampsia (PE) is a human pregnancy-specific syndrome with abnormal activation of cells from the innate immune system. The present study evaluated whether silibinin (SB) treatment of monocytes from preeclamptic women could modulate NLRP1 and NLRP3 inflammasomes as well as TLR4/NF-κB pathway activation. Peripheral blood monocytes from 20 preeclamptic and 20 normotensive (NT) pregnant women, as well as the THP-1 cell line, were cultured with or without monosodium urate (MSU) or SB. NLRP1, NLRP3, Caspase-1, TLR4, MyD88, NF-κB, IL-1β, IL-18, TNF-α and IL-10 gene expression by monocytes was analysed by quantitative real-time polymerase chain reaction (qPCR), while inflammatory cytokine production and p65NF-κB activity were determined by enzyme-linked immunosorbent assays (ELISAs). TLR4/MyD88/NF-κB and NLRP1/NLRP3 inflammasomes pathways in THP-1 cells were evaluated by flow cytometry and western blot respectively. Compared with NT women, monocytes from preeclamptic women showed The Ethics Committee of the Botucatu Medical School approved the study (protocol number 2.333.216)higher endogenous activation of NLRP1/NLRP3 inflammasomes and the TLR4/NF-κB pathway as well as higher gene and protein expression of IL-1β, IL-18 and TNF-α, and lower expression of IL-10. Monocyte stimulation with MSU increased inflammation-related genes as well as NF-κB activity. In vitro, SB treatment of monocytes from preeclamptic women reduced the basal activation of these cells by decreasing NLRP1/NLRP3 inflammasomes and p65NF-κB activity. THP-1 cells exhibited a similar immunological response profile to monocytes from preeclamptic women when cultured with or without MSU or SB. These results suggest uric acid participates in the systemic inflammatory response characteristic of preeclampsia and that in vitro SB treatment can modulate the sterile inflammation established in monocytes from preeclamptic women.

Upregulation of the thioredoxin-dependent redox system during differentiation of 3T3-L1 cells to adipocytes

2014 ◽  
Vol 395 (6) ◽  
pp. 667-677 ◽  
Author(s):  
Ann-Marie Rajalin ◽  
Mustafa Micoogullari ◽  
Helmut Sies ◽  
Holger Steinbrenner
Keyword(s):  
Hydrogen Peroxide ◽  
Redox System ◽  
Antioxidant Systems ◽  
Protein Levels ◽  
Glutathione Peroxidases ◽  
Tnf Α ◽  
Gene And Protein Expression ◽  
Hydrogen Peroxide Generation ◽  
Factor Α ◽  
Thioredoxin Reductases

Abstract Hydrogen peroxide acts as a signaling molecule in early adipogenesis. In differentiating adipocytes, elevated hydrogen peroxide generation is balanced through induction of antioxidant enzymes such as catalase and peroxiredoxins. Thioredoxin reductases (TrxR) and glutathione peroxidases (GPx) are selenoenzymes that constitute part of the major thiol-dependent antioxidant systems in cells. Here we show that the protein levels of cytoplasmic/nuclear TrxR1 and mitochondrial TrxR2 increase in the course of adipocyte differentiation of 3T3-L1 cells together with the TrxR2 substrate thioredoxin 2 (Trx2), resulting in elevated TrxR activity in mature adipocytes. Gene and protein expression of the GPx isoenzyme GPx4 was also stimulated during adipogenesis. Chronic exposure of 3T3-L1 cells to the anti-adipogenic factors tumor necrosis factor α (TNF-α) or rapamycin during differentiation suppressed TrxR1 and Trx2 upregulation, concomitantly with inhibition of adipogenesis and lipogenesis. In contrast, TNF-α or rapamycin did not affect expression of TrxRs and their Trx substrates in mature adipocytes. These results indicate that upregulation of the thioredoxin-dependent redox system is linked to the development of an adipocyte phenotype.

PAR2-induced inflammatory responses in human kidney tubular epithelial cells

2013 ◽  
Vol 304 (6) ◽  
pp. F737-F750 ◽  
Author(s):  
David A. Vesey ◽  
Jacky Y. Suen ◽  
Vernon Seow ◽  
Rink-Jan Lohman ◽  
Ligong Liu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Inflammatory Responses ◽  
Cell Monolayer ◽  
Human Kidney ◽  
Tubular Epithelial Cells ◽  
Pronounced Increase ◽  
Gm Csf ◽  
Tnf Α ◽  
Gene And Protein Expression

Protease-activated receptor-2 (PAR2) is a G protein-coupled receptor abundantly expressed in the kidney. The aim of this study was to profile inflammatory gene and protein expression induced by PAR2 activation in human kidney tubular epithelial cells (HTEC). A novel PAR2 antagonist, GB88, was used to confirm agonist specificity. Intracellular Ca2+ (iCa2+) mobilization, confocal microscopy, gene expression profiling, qRTPCR, and protein expression were used to characterize PAR2 activation. PAR2 induced a pronounced increase in iCa2+ concentration that was blocked by the PAR2 antagonist. Treatment with SLIGKV-NH2 at the apical or basolateral cell surface for 5 h induced expression of a range of inflammatory genes by greater than fourfold, including IL-1β, TRAF1, IL-6, and MMP-1, as assessed by cDNA microarray and qRTPCR analysis. Using antibody arrays, GM-CSF, ICAM-1, TNF-α, MMP-1, and MMP-10 were among the induced proteins secreted. Cytokine-specific ELISAs identified three- to sixfold increases in GM-CSF, IL-6, IL-8, and TNF-α, which were blocked by GB88 and protein kinase C inhibitors. Treatment of cells at the basolateral surface induced more potent inflammatory responses, with release of MCP-1 and fibronectin to the apical and basolateral compartments; apical treatment only increased secretion of these factors to the apical compartment. PAR2 activation at the basolateral surface dramatically reduced transepithelial electrical resistance (TEER) whereas apical treatment had no effect. There was very little leakage (<5%) of peptides across the cell monolayer (liquid chromatography-mass spectrometry). In summary, SLIGKV-NH2 induced robust proinflammatory responses in HTEC that were antagonized by GB88. These results suggest that PAR2 antagonists could be useful disease-modifying, anti-inflammatory agents in kidney disease.

scholarly journals Enhanced Cerebral Expression of MCT1 and MCT2 in a Rat Ischemia Model Occurs in Activated Microglial Cells

2009 ◽  
Vol 29 (7) ◽  
pp. 1273-1283 ◽  
Author(s):  
Tiago JTP Moreira ◽  
Karin Pierre ◽  
Fumihiko Maekawa ◽  
Cendrine Repond ◽  
Aleta Cebere ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna Level ◽  
Protein Levels ◽  
Microglial Cell Line ◽  
Tnf Α ◽  
Ischemic Episode ◽  
Isolectin B4 ◽  
Factor Α ◽  
Necrosis Factor

Monocarboxylate transporters (MCTs) are essential for the use of lactate, an energy substrate known to be overproduced in brain during an ischemic episode. The expression of MCT1 and MCT2 was investigated at 48 h of reperfusion from focal ischemia induced by unilateral extradural compression in Wistar rats. Increased MCT1 mRNA expression was detected in the injured cortex and hippocampus of compressed animals compared to sham controls. In the contralateral, uncompressed hemisphere, increases in MCT1 mRNA level in the cortex and MCT2 mRNA level in the hippocampus were noted. Interestingly, strong MCT1 and MCT2 protein expression was found in peri-lesional macrophages/microglia and in an isolectin B4+/S100β+ cell population in the corpus callosum. In vitro, MCT1 and MCT2 protein expression was observed in the N11 microglial cell line, whereas an enhancement of MCT1 expression by tumor necrosis factor-α (TNF-α) was shown in these cells. Modulation of MCT expression in microglia suggests that these transporters may help sustain microglial functions during recovery from focal brain ischemia. Overall, our study indicates that changes in MCT expression around and also away from the ischemic area, both at the mRNA and protein levels, are a part of the metabolic adaptations taking place in the brain after ischemia.

Mechanisms of MAdCAM-1 gene expression in human intestinal microvascular endothelial cells

2005 ◽  
Vol 288 (2) ◽  
pp. C272-C281 ◽  
Author(s):  
Hitoshi Ogawa ◽  
David G. Binion ◽  
Jan Heidemann ◽  
Monica Theriot ◽  
Pamela J. Fisher ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Protein Expression ◽  
Molecular Mechanisms ◽  
Primary Cultures ◽  
Microvascular Endothelial Cells ◽  
Cellular Density ◽  
Tnf Α ◽  
Gene And Protein Expression ◽  
Microvascular Endothelial

Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a homing receptor preferentially expressed on gut-associated endothelial cells that plays a central role in leukocyte traffic into the mucosal immune compartment. Although the molecular mechanisms underlying endothelial ICAM-1 or E-selectin expression have been intensively investigated, the mechanisms that regulate human MAdCAM-1 expression have not been defined. We report MAdCAM-1 gene and protein expression in primary cultures of human intestinal microvascular endothelial cells (HIMEC) that was not demonstrated in human umbilical vein endothelial cells. Similar to ICAM-1 and E-selectin expression, MAdCAM-1 gene expression in HIMEC was inducible with TNF-α, IL-1β, or LPS activation. However, in striking contrast to ICAM-1 and E-selectin expression, MAdCAM-1 mRNA and protein expression in HIMEC was heavily dependent on culture duration and/or cellular density, suggesting a prominent role for cell-cell interaction among these endothelial cells in the expression of the mucosal addressin. MAdCAM-1 expression was inhibited by both SN-50 (NF-κB inhibitor) and LY-294002 [phosphatidylinositol 3-kinase (PI3-K) inhibitor], whereas ICAM-1 and E-selectin expression was inhibited by SN-50 but not by LY-294002. The Akt phosphorylation by TNF-α or LPS was greater at higher cell density, demonstrating a pattern similar to that of MAdCAM-1 expression. NF-κB activation was not affected by cellular density in HIMEC. MAdCAM-1 expression in human gut endothelial cells is regulated by distinct signaling mechanisms involving both NF-κB and PI3-K/Akt. These data also suggest that PI3-K/Akt is involved in the gut-specific differentiation of HIMEC, which results in expression of the mucosal addressin MAdCAM-1.

scholarly journals LIPUS inhibits inflammation and catabolism through the NF‐κB pathway in human degenerative nucleus pulposus cells

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Weiwei Yi ◽  
Qing Chen ◽  
Chuan Liu ◽  
Kaiting Li ◽  
Bailong Tao ◽  
...  
Keyword(s):  
Protein Expression ◽  
Real Time ◽  
Signaling Pathway ◽  
Nucleus Pulposus ◽  
Real Time Pcr ◽  
Inflammatory Factors ◽  
Nucleus Pulposus Cells ◽  
Tnf Α ◽  
Gene And Protein Expression ◽  
Collagen Ii

Abstract Background Low-intensity pulsed ultrasound (LIPUS) is a safe and noninvasive rehabilitative physical therapy with anti-inflammatory effects. The current study investigated the effect of LIPUS on the inflammation of nucleus pulposus (NP) cells and its underlying mechanism. Methods Human NP cells were acquired from lumbar disc herniation tissue samples and cultured for experiments. Human NP cells were treated with LPS and then exposed to LIPUS (15 mW/cm2, 30 mW/cm2 and 60 mW/cm2) for 20 min daily for 3 days to determine the appropriate intensity to inhibit the expression of the inflammatory factors TNF-α and IL-1β. The gene and protein expression of aggrecan, collagen II, MMP-3 and MMP-9 was measured by real‐time PCR and western blotting, respectively. The activity of the nuclear factor‐kappa B (NF‐κB) pathway was examined by western blotting and immunofluorescence. After pretreatment with the NF-κB inhibitor PDTC, the expression of TNF-α, IL-1β, MMP-3 and MMP-9 was measured by real‐time PCR. Results LIPUS at intensities of 15 mW/cm2, 30 mW/cm2 and 60 mW/cm2 inhibited LPS-induced NP cell expression of the inflammatory factors TNF-α and IL-1β, especially at 30 mW/cm2. LIPUS significantly upregulated the gene and protein expression of aggrecan and collagen II and downregulated the gene and protein expression of MMP-3 and MMP-9 in LPS-induced NP cells. The NF‐κB signaling pathway was inhibited by LIPUS through inhibiting the protein expression of p-P65 and the translocation of P65 into the nucleus in LPS-induced NP cells. In addition, LIPUS had similar effects as the NF-κB inhibitor PDTC by inhibiting the NF-κB signaling pathway, inflammation and catabolism in LPS-induced human degenerative nucleus pulposus cells. Conclusion LIPUS inhibited inflammation and catabolism through the NF‐κB pathway in human degenerative nucleus pulposus cells.

scholarly journals Qualitative detection of haptoglobin mRNA in bovine and human blood leukocytes and bovine milk somatic cells

2012 ◽  
Vol 50 (No. 12) ◽  
pp. 515-520
Author(s):  
Thielen MA ◽  
M. Mielenz ◽  
S. Hiss ◽  
H. Sauerwein
Keyword(s):  
Mammary Gland ◽  
Somatic Cells ◽  
Bovine Milk ◽  
Bovine Mammary Gland ◽  
Blood Leukocytes ◽  
Human Leukocytes ◽  
Mrna Transcripts ◽  
Stromal Tissue ◽  
Tissue Homogenates ◽  
Milk Somatic Cells

Local expression of haptoglobin (Hp) mRNA in tissue homogenates from different sites of the bovine mammary gland has recently been established. Besides alveolar and stromal tissue, blood cells are one potential cellular location of Hp expression. The aim of this study was to investigate whether bovine leukocytes might contribute to the Hp mRNA transcripts found in the mammary gland after their migration into this organ. For this purpose RT-PCR was carried out with total RNA extracted from leukocytes in blood and from somatic cells in milk of three dairy cows. In comparison, human leukocytes of four donors were examined. Both bovine blood and milk somatic cells showed distinct signals for Hp mRNA in all samples in contrast to human leukocytes where Hp mRNA was detected in only two out of four donors after reamplification. In conclusion, the presence of Hp mRNA in blood derived leukocytes indicates that these cells contribute to the Hp mRNA transcripts found in the bovine mammary gland. It remains to be clarified whether other cells within the mammary gland are also capable of Hp mRNA synthesis.

scholarly journals Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells

2020 ◽  
Vol 295 (33) ◽  
pp. 11529-11541 ◽  
Author(s):  
Sarah M. Gray ◽  
Yurong Xin ◽  
Elizabeth C. Ross ◽  
Bryanna M. Chazotte ◽  
Megan E. Capozzi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Islet Cells ◽  
Expression Patterns ◽  
Metabolic Stress ◽  
Β Cells ◽  
Β Cell ◽  
Protein Levels ◽  
Gene And Protein Expression ◽  
Glucagon Like Peptide 1 ◽  
Mouse Pancreatic Islet

The insulinotropic actions of glucagon-like peptide 1 receptor (GLP-1R) in β-cells have made it a useful target to manage type 2 diabetes. Metabolic stress reduces β-cell sensitivity to GLP-1, yet the underlying mechanisms are unknown. We hypothesized that Glp1r expression is heterogeneous among β-cells and that metabolic stress decreases the number of GLP-1R–positive β-cells. Here, analyses of publicly available single-cell RNA-Seq sequencing (scRNASeq) data from mouse and human β-cells indicated that significant populations of β-cells do not express the Glp1r gene, supporting heterogeneous GLP-1R expression. To check these results, we used complementary approaches employing FACS coupled with quantitative RT-PCR, a validated GLP-1R antibody, and flow cytometry to quantify GLP-1R promoter activity, gene expression, and protein expression in mouse α-, β-, and δ-cells. Experiments with Glp1r reporter mice and a validated GLP-1R antibody indicated that >90% of the β-cells are GLP-1R positive, contradicting the findings with the scRNASeq data. α-cells did not express Glp1r mRNA and δ-cells expressed Glp1r mRNA but not protein. We also examined the expression patterns of GLP-1R in mouse models of metabolic stress. Multiparous female mice had significantly decreased β-cell Glp1r expression, but no reduction in GLP-1R protein levels or GLP-1R–mediated insulin secretion. These findings suggest caution in interpreting the results of scRNASeq for low-abundance transcripts such as the incretin receptors and indicate that GLP-1R is widely expressed in β-cells, absent in α-cells, and expressed at the mRNA, but not protein, level in δ-cells.

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