Effect of DNA Methylation on LPS-Induced Expression of Tumour Necrosis Factor Alpha (TNF-α) in Bovine Mammary Epithelial Cells

2021 ◽  
Author(s):  
Yangyunyi Dong ◽  
Dong An ◽  
Jun Wang ◽  
Hongyu Liu ◽  
Qing Zhang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Tumour Necrosis Factor Alpha ◽  
Mammary Epithelial ◽  
Necrosis Factor Alpha ◽  
Bovine Mammary Epithelial Cells ◽  
Tnf Α ◽  
Factor Alpha ◽  
Necrosis Factor

Background: Cow mastitis is a major disease that affects dairy industry worldwide. Although the inflammatory response induced by lipopolysaccharide (LPS) in bovine mammary epithelial cells (BMECs) is similar to the response to pathogenic bacteria, the underlying regulatory mechanisms remain unclear. This study aimed to clarify the effect of DNA methylation on LPS-induced expression of tumour necrosis factor alpha (TNF-α) in BMECs. Methods: The mammary epithelial cells were treated with LPS and DNA methylation inhibition 5-Aza-2’-deoxycytodine (5Aza). Expression of TNF-α, IL-6, BNBD-5, DNA methyltransferases (DNMT1, DNMT2, DNMT3A and DNMT3B) and DNA methylation at TNF-α regions, were analyzed using quantitative realtime PCR (qRT-PCR), Elisa and bisulfite sequencing PCR (BSP). Result: Our results showed that LPS significantly increased the expression of inflammatory factors, including interleukin-6 (IL-6), bovine neutrophil beta-defensins (BNBD-5) and TNF-α. Further, we observed that the DNA methylation inhibitor, 5-Aza-2'-deoxycytosine (5-Aza), enhanced LPS-induced TNF-α mRNA expression. In addition, we found that LPS treatment significantly decreased the methylation levels of specific CpG sites in the TNF-α promoter (at -245 and -323) and inhibited the expression of DNA methyl transferases (DNMT1, DNMT2, DNMT3A and DNMT3B). Our results indicate that LPS promotes the expression of TNF-α in BMECs by inhibiting DNA methylation in the gene promoter region.

scholarly journals Comparative Kinetics ofEscherichia coli- andStaphylococcus aureus-Specific Activation of Key Immune Pathways in Mammary Epithelial Cells Demonstrates ThatS. aureusElicits a Delayed Response Dominated by Interleukin-6 (IL-6) but Not by IL-1A or Tumor Necrosis Factor Alpha

2010 ◽  
Vol 79 (2) ◽  
pp. 695-707 ◽  
Author(s):  
Juliane Günther ◽  
Kathrin Esch ◽  
Norbert Poschadel ◽  
Wolfram Petzl ◽  
Holm Zerbe ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis Factor Alpha ◽  
Tumor Necrosis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Necrosis Factor Alpha ◽  
E Coli ◽  
Tnf Α ◽  
Factor Alpha ◽  
Necrosis Factor

ABSTRACTInfections of the udder byEscherichia colivery often elicit acute inflammation, whileStaphylococcus aureusinfections tend to cause mild, subclinical inflammation and persistent infections. The molecular causes underlying the different disease patterns are poorly understood. We therefore profiled the kinetics and extents of global changes in the transcriptome of primary bovine mammary epithelial cells (MEC) after challenging them with heat-inactivated preparations ofE. coliorS. aureuspathogens.E. coliswiftly and strongly induced an expression of cytokines and bactericidal factors.S. aureuselicited a retarded response and failed to quickly induce an expression of bactericidal factors. Both pathogens induced similar patterns of chemokines for cell recruitment into the udder, butE. colistimulated their synthesis much faster and stronger. The genes that are exclusively and most strongly upregulated byE. colimay be clustered into a regulatory network with tumor necrosis factor alpha (TNF-α) and interleukin-1 (IL-1) in a central position. In contrast, the expression of these master cytokines is barely regulated byS. aureus. Both pathogens quickly trigger an enhanced expression of IL-6. This is still possible after completely abrogating MyD88-dependent Toll-like receptor (TLR) signaling in MEC. TheE. coli-specific strong induction of TNF-α and IL-1 expression may be causative for the severe inflammatory symptoms of animals suffering fromE. colimastitis, while the avoidance to quickly induce the synthesis of bactericidal factors may support the persistent survival ofS. aureuswithin the udder. We suggest thatS. aureussubverts the MyD88-dependent activation of immune gene expression in MEC.

Tumor necrosis factor alpha induces LIF expression through ERK1/2 activation in mammary epithelial cells

2010 ◽  
Vol 110 (4) ◽  
pp. 857-865 ◽  
Author(s):  
Carolina Schere Levy ◽  
Victoria Slomiansky ◽  
Albana Gattelli ◽  
Karen Nahmod ◽  
Federico Pelisch ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Epithelial Cells ◽  
Tumor Necrosis Factor Alpha ◽  
Tumor Necrosis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Necrosis Factor Alpha ◽  
Factor Alpha ◽  
Necrosis Factor

scholarly journals MiR-125b regulates inflammation in bovine mammary epithelial cells by targeting the NKIRAS2 gene

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Zhuo-Ma Luoreng ◽  
Da-Wei Wei ◽  
Xing-Ping Wang
Keyword(s):  
T Cells ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Luciferase Reporter ◽  
Regulation Mechanism ◽  
Bovine Mammary Epithelial Cells ◽  
Tnf Α

AbstractMastitis is a complex inflammatory disease caused by pathogenic infection of mammary tissue in dairy cows. The molecular mechanism behind its occurrence, development, and regulation consists of a multi-gene network including microRNA (miRNA). Until now, there is no report on the role of miR-125b in regulating mastitis in dairy cows. This study found that miR-125b expression is significantly decreased in lipopolysaccharide (LPS)-induced MAC-T bovine mammary epithelial cells. Also, its expression is negatively correlated with the expression of NF-κB inhibitor interacting Ras-like 2 (NKIRAS2) gene. MiR-125b target genes were identified using a double luciferase reporter gene assay, which showed that miR-125b can bind to the 3′ untranslated region (3′ UTR) of the NKIRAS2, but not the 3′UTR of the TNF-α induced protein 3 (TNFAIP3). In addition, miR-125b overexpression and silencing were used to investigate the role of miR-125b on inflammation in LPS-induced MAC-T. The results demonstrate that a reduction in miR-125b expression in LPS-induced MAC-T cells increases NKIRAS2 expression, which then reduces NF-κB activity, leading to low expression of the inflammatory factors IL-6 and TNF-α. Ultimately, this reduces the inflammatory response in MAC-T cells. These results indicate that miR-125b is a pro-inflammatory regulator and that its silencing can alleviate bovine mastitis. These findings lay a foundation for elucidating the molecular regulation mechanism of cow mastitis.

Tea tree oil prevents mastitis-associated inflammation in lipopolysaccharide-stimulated bovine mammary epithelial cells

2020 ◽  
Author(s):  
Zhi Chen ◽  
Yi Zhang ◽  
Jingpeng Zhou ◽  
Yu Tian ◽  
Qiaoni Zhou ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Differentially Expressed ◽  
Quantitative Detection ◽  
Tea Tree Oil ◽  
Tea Tree ◽  
Bovine Mammary Epithelial Cells ◽  
Tnf Α ◽  
Prevention And Treatment

Abstract Background Effective prevention and treatment of cow mastitis can provide a good guarantee for the healthy growth of cows and the qualified production of dairy products. The main purpose of this study was to explore the effect of tea tree oil on lipopolysaccharide (LPS) -induced mastitis in dairy cows, and the key gene in LPS -stimulated bovine mammary epithelial cells (BMECs) was identified. Results In this study, a model of mastitis induced by LPS was constructed, to which tea tree oil and LPS were added. The protective effects of tea tree oil on LPS-induced mastitis in BMECs were verified by the CCK-8 method, flow cytometry, real-time fluorescence quantitative detection, ELISA and other methods. The results showed that LPS at a concentration of 200 μg/ml could reduce the proliferative activity of the cells, induce a high proportion of apoptosis, and promote the expression of TNF-α, IL-6 and STAT1. Upon addition of tea tree oil, the proportion of apoptosis was reduced, and the expression of NF-κB, MAPK and caspase-3 was inhibited. Mammary epithelial cells were compared under control and LPS-treatment conditions and analyzed by second-generation sequencing. A total of 1270 mRNAs were identified as differentially expressed, of which 787 genes were upregulated and 483 were downregulated. These differentially expressed genes include TNF - α, IL6, STAT1, mapk4, etc. H&E staining and immunohistochemistry were used to verify the function of candidate genes. TNF-α and IL6 were observed to play important roles in mediating the preventive effect of tea tree oil on mastitis in LPS-stimulated bovine mammary epithelial cells. Conclusions The results showed that tea tree oil had a protective effect against LPS-induced mastitis. TNF - α and IL6 may be the marker genes of LPS-induced mastitis which provided a theoretical basis and experimental support for further research to determine new strategies for the prevention and treatment of mastitis and improvement of milk quality.

scholarly journals Isolated Flavonoids against Mammary Tumour Cells LM2

2009 ◽  
Vol 64 (1-2) ◽  
pp. 32-36 ◽  
Author(s):  
Camila B. de A. Carli ◽  
Djamile C. de Matos ◽  
Flávia C. M. Lopes ◽  
Danielle C. G. Maia ◽  
Maristela B. Dias ◽  
...  
Keyword(s):  
Antitumour Activity ◽  
Tumour Necrosis Factor Alpha ◽  
Tumour Cells ◽  
Mammary Tumour ◽  
Sandwich Assay ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Factor Alpha ◽  
Almost All ◽  
Necrosis Factor

1The purpose of the present study was to investigate antitumour and anti-inflammatory activities of flavonoids isolated from Byrsonima crassa, Davilla elliptica and Mouriri pusa. The antitumour activity was measured by the MTT assay in murine mammary tumour cells (LM2) and the IC50 values of the fl avonoids tested ranged from (31.5 ± 2.97) to (203.1 ± 5.9) μg/ml. The fl avonoids (myricetin-3-O-α-L-rhamnopyranoside) and 3 (quercetin-3-Ogalactopyranoside) from D. elliptica were the most active ones against the tumour cells. The same samples were tested to determine the inhibition of the release of nitric oxide (NO) and of the tumour necrosis factor-alpha (TNF-α) in murine macrophages by the Griess and ELISA sandwich assay, respectively. Almost all the samples showed inhibitory activity to the release of NO but not of TNF-α. Of all substances tested, flavonoids 2 (quercetin) and 6 (myricetin) may show promising activity in the treatment of murine breast cancer by immunomodulatory and antiproliferative activities.

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