MicroRNA-141 participates in milk lipid metabolism by targeting SIRT1 in bovine mammary epithelial cells

2020 ◽  
Vol 60 (16) ◽  
pp. 1877
Author(s):  
Yujia Sun ◽  
Hailei Xia ◽  
Xubin Lu ◽  
Chong Xu ◽  
Mingxun Li ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Luciferase Assay ◽  
Milk Lipid ◽  
Potential Candidate ◽  
Bovine Mammary Epithelial Cells ◽  
Sirt1 Gene

Context The regulation of milk lipids is important for the evaluation of dairy cows’ performance. Lipids are produced and secreted by mammary gland under the regulation of steroid hormones, growth factors and microRNAs (miRNAs). MicroRNAs have been verified to be involved in numerous biological processes. Previous studies have shown that miR-141 is expressed at higher levels in dairy cows at peak lactation than in those at early lactation. However, the roles of miR-141 in bovine mammary epithelial cells (BMECs) and the mechanisms how it affects lipid metabolism are as yet unknown. Aims The aims of this study were to clarify (i) the molecular mechanisms of miR-141 in milk lipid metabolism, and (ii) how miR-141 affects milk lipid metabolism in BMECs. Methods Triglycerides were observed in BMECs using triglyceride analysis after overexpression or inhibition of miR-141; selected potential candidate genes that are targeted by miR-141 using TargetScan. The regulatory relationship among miR-141, SIRT1 gene and lipid metabolism-related genes (SREBF1, FASN and PPARγ) by using the dual luciferase assay, quantitative real-time PCR and western blotting. Key results Through overexpression or inhibition of miR-141 expression, we found that miR-141 promoted lipid metabolism in BMECs and an increase in triglycerides was observed in these cells. Further, miR-141 targets the 3′UTR of SIRT1 mRNA, and negative regulates the expression of SIRT1 gene in BMECs. Also, the expression levels of SREBF1, FASN and PPARγ, which are related to milk lipid metabolism, were also altered after overexpression miR-141. Conclusions Our results have revealed that miR-141 could promote milk lipid metabolism in BMECs by means of negative regulates SIRT1 gene and positive effects lipid metabolism-related genes (SREBF1, FASN and PPARγ) in BMECs. Implications Our research indicates that miR-141 could be considered a marker in cattle breeding to obtain high quality dairy products. It would be useful to study the function of miRNAs in milk lipid metabolism and synthesis. In the long term these findings might be helpful in developing practical means to improve the quality of ruminant milk.

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.

scholarly journals Circadian Gene PER2 Silencing Downregulates PPARG and SREBF1 and Suppresses Lipid Synthesis in Bovine Mammary Epithelial Cells

Biology ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1226
Author(s):  
Yujia Jing ◽  
Yifei Chen ◽  
Shan Wang ◽  
Jialiang Ouyang ◽  
Liangyu Hu ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Epithelial Cells ◽  
Circadian Clock ◽  
Lipid Droplets ◽  
Mammary Epithelial Cells ◽  
Lipid Synthesis ◽  
Mammary Epithelial ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Bovine Mammary Epithelial Cells

PER2, a circadian clock gene, is associated with mammary gland development and lipid synthesis in rodents, partly via regulating peroxisome proliferator-activated receptor gamma (PPARG). Whether such a type of molecular link existed in bovines was unclear. We hypothesized that PER2 was associated with lipid metabolism and regulated cell cycles and apoptosis in bovine mammary epithelial cells (BMECs). To test this hypothesis, BMECs isolated from three mid-lactation (average 110 d postpartum) cows were used. The transient transfection of small interfering RNA (siRNA) was used to inhibit PER2 transcription in primary BMECs. The silencing of PER2 led to lower concentrations of cellular lipid droplets and triacylglycerol along with the downregulation of lipogenic-related genes such as ACACA, FASN, LPIN1, and SCD, suggesting an overall inhibition of lipogenesis and desaturation. The downregulation of PPARG and SREBF1 in response to PER2 silencing underscored the importance of circadian clock signaling and the transcriptional regulation of lipogenesis. Although the proliferation of BMECs was not influenced by PER2 silencing, the number of cells in the G2/GM phase was upregulated. PER2 silencing did not affect cell apoptosis. Overall, the data provided evidence that PER2 participated in the coordination of mammary lipid metabolism and was potentially a component of the control of lipid droplets and TAG synthesis in ruminant mammary cells. The present data suggested that such an effect could occur through direct effects on transcriptional regulators.

Transcriptional regulation of milk lipid synthesis by exogenous C16:0 and C18 fatty acids in bovine mammary epithelial cells

2018 ◽  
Vol 98 (2) ◽  
pp. 260-270 ◽  
Author(s):  
Ni Dan ◽  
Hang Zhang ◽  
Changjin Ao ◽  
Khas-Erdene
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Regulatory Element ◽  
Fatty Acid Synthetase ◽  
Mammary Epithelial ◽  
Lower Amount ◽  
Milk Lipid ◽  
Bovine Mammary Epithelial Cells

The objective of this study was to examine the effects of removing one fatty acid from a combination of long-chain fatty acids (LCFA) on milk lipogenesis in bovine mammary epithelial cells. The incubation concentration of LCFA was determined, and 100 μmol L−1 of C16:0, 5 μmol L−1 of C18:0, 100 μmol L−1 of cis-9 C18:1, 25 μmol L−1 of n-6 C18:2, and 1.2 μmol L−1 of n-3 C18:3 were used in the study. Treatments were C16:0, C18:0, C18:1, C18:2, and C18:3 combinations as control; control absent of C16:0 as A-C16:0; control absent of C18:0 as A-C18:0; control absent of C18:1 as A-C18:1; control absent of C18:2 as A-C18:2; control absent of C18:3 as A-C18:3. Results showed that compared with control, fatty acid synthetase expression was reduced by A-C18:0 and A-C18:1. Palmitic acid decreased expression of lipoprotein lipase. Compared with control, the expression of stearoyl-coenzyme A desaturase-1 and cluster of differentiation 36 was reduced by all treatments. Peroxisome proliferator-activated receptor-α expression was down-regulated by A-C16:0, A-C18:0, A-C18:1, and A-C18:2. Sterol regulatory element binding factor-1 was decreased when treated with A-C18:0, A-C18:1, and A-C18:2. Cells lack of 18-carbon fatty acid synthesized lower amount of intracellular triglyceride compared with control.

scholarly journals Beta-Sitosterol Promotes Milk Protein and Fat Syntheses-Related Genes in Bovine Mammary Epithelial Cells

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3238
Author(s):  
Xinlu Liu ◽  
Jinglin Shen ◽  
Jinxin Zong ◽  
Jiayi Liu ◽  
Yongcheng Jin
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Dairy Cows ◽  
Milk Fat ◽  
Milk Protein ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells ◽  
Mrna And Protein Expression ◽  
Fat Synthesis

β-sitosterol, a phytosterol with multiple biological activities, has been used in the pharmaceutical industry. However, there are only a few reports on the use of β-sitosterol in improving milk synthesis in dairy cows. This study aimed to investigate the effects of β-sitosterol on milk fat and protein syntheses in bovine mammary epithelial cells (MAC-T) and its regulatory mechanism. MAC-T cells were treated with different concentrations (0.01, 0.1, 1, 5, 10, 20, 30, or 40 μM) of β-sitosterol, and the expression levels of milk protein and fat synthesis-related genes and proteins were analyzed. β-sitosterol at 0.1, 1, and 10 μM concentrations promoted the mRNA and protein expression of β-casein. β-sitosterol (0.1, 1, 10 μM) increased the mRNA and protein expression levels of signal transducer activator of transcription 5 (STAT5), mammalian target of rapamycin (mTOR), and ribosomal protein S6 kinase beta-1 (S6K1) of the JAK2/STAT5 and mTOR signaling pathways. It also stimulated the milk fat synthesis-related factors, including sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor-gamma (PPARγ), acetyl-CoA carboxylase (ACC), lipoprotein lipase (LPL), and stearyl CoA desaturase (SCD). β-sitosterol (0.1, 1, 10 μM) also significantly increased the expression of growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis and hypoxia-inducible factor-1α (HIF-1α)-related genes. Notably, the compound inhibited the expression of the negative regulator, the suppressor of cytokine signaling 2 (SOCS2) at the two lower concentrations (0.1, 1 μM), but significantly promoted the expression at the highest concentration (30 μM). These results highlight the role of β-sitosterol at concentrations ranging from 0.1 to 10 μM in improving milk protein and fat syntheses, regulating milk quality. Therefore, β-sitosterol can be used as a potential feed additive to improve milk quality in dairy cows.

scholarly journals PRL/microRNA-183/IRS1 Pathway Regulates Milk Fat Metabolism in Cow Mammary Epithelial Cells

Genes ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 196 ◽  
Author(s):  
Peixin Jiao ◽  
Yuan Yuan ◽  
Meimei Zhang ◽  
Youran Sun ◽  
Chuanzi Wei ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Molecular Mechanism ◽  
Milk Fat ◽  
Mammary Epithelial Cells ◽  
Cpg Island ◽  
Fat Metabolism ◽  
Milk Quality ◽  
Mammary Epithelial

The aim of the study was to understand the internal relationship between milk quality and lipid metabolism in cow mammary glands. A serial of studies was conducted to assess the molecular mechanism of PRL/microRNA-183/IRS1 (Insulin receptor substrate) pathway, which regulates milk fat metabolism in dairy cows. microRNA-183 (miR-183) was overexpressed and inhibited in cow mammary epithelial cells (CMECs), and its function was detected. The function of miR-183 in inhibiting milk fat metabolism was clarified by triglycerides (TAG), cholesterol and marker genes. There is a CpG island in the 5′-flanking promoter area of miR-183, which may inhibit the expression of miR-183 after methylation. Our results showed that prolactin (PRL) inhibited the expression of miR-183 by methylating the 5′ terminal CpG island of miR-183. The upstream regulation of PRL on miR-183 was demonstrated, and construction of the lipid metabolism regulation network of microRNA-183 and target gene IRS1 was performed. These results reveal the molecular mechanism of PRL/miR-183/IRS1 pathway regulating milk fat metabolism in dairy cows, thus providing an experimental basis for the improvement of milk quality.

scholarly journals New function of the CD44 gene: Lipid metabolism regulation in bovine mammary epithelial cells

2020 ◽  
Vol 103 (7) ◽  
pp. 6661-6671 ◽  
Author(s):  
Ping Jiang ◽  
Lixin Xia ◽  
ZiKang Jin ◽  
Shaokat Ali ◽  
Mengyan Wang ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells ◽  
Metabolism Regulation

Effects of exogenous C18 unsaturated fatty acids on milk lipid synthesis in bovine mammary epithelial cells

2020 ◽  
Vol 87 (3) ◽  
pp. 344-348
Author(s):  
Hang Zhang ◽  
Ni Dan ◽  
Changjin Ao ◽  
Sizhen Wang ◽  
Khas Erdene ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Epithelial Cells ◽  
Unsaturated Fatty Acids ◽  
Mammary Epithelial Cells ◽  
Lipid Synthesis ◽  
Mammary Epithelial ◽  
Control Treatment ◽  
Milk Lipid ◽  
Bovine Mammary Epithelial Cells

AbstractWe determined the effects of a combination of C18 unsaturated fatty acids (C18-UFAs) consisting of oleic, linoleic, and linolenic acids on milk lipogenesis in bovine mammary epithelial cells (BMECs). By orthogonal experiments to determine cellular triacylglycerol (TAG) accumulation, a combination of 200 μmol/l C18 : 1, 50 μmol/l C18 : 2, and 2 μmol/l C18 : 3 was selected as C18-UFAs combination treatment, and culture in medium containing fatty acid-free bovine serum albumin was used as the control. The expression of genes related to milk lipid synthesis and intracellular FA composition was measured. The results showed that cytosolic TAG formation was higher under C18-UFAs treatment than under control treatment. The mRNA expression of acetyl-CoA carboxylase-α (ACACA), fatty acid synthase (FASN), and peroxisome proliferator-activated receptor gamma (PPARG) did not differ between treatments. The abundance of stearoyl-CoA desaturase (SCD) and acyl-CoA synthetase long-chain family member 1 (ACSL1) was higher, whereas that of sterol regulatory element binding transcription factor 1 (SREBF-1) was lower after C18-UFAs treatment compared to control treatment. The C16 : 0 and SFA content was decreased following C18-UFAs treatment compared to control treatment, while the cis-9 C18 : 1 and UFA content was increased. In conclusion, C18-UFAs could stimulate triglyceride accumulation, increase the cellular UFA concentration, and regulate lipogenic genes in BMECs.

scholarly journals Astragaloside IV Alleviates Ammonia-Induced Apoptosis and Oxidative Stress in Bovine Mammary Epithelial Cells

2019 ◽  
Vol 20 (3) ◽  
pp. 600 ◽  
Author(s):  
Fengge Wang ◽  
Yun Zhao ◽  
Shuxiong Chen ◽  
Lu Chen ◽  
Liting Sun ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
T Cells ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Inflammatory Responses ◽  
Mammary Epithelial Cells ◽  
Mammary Glands ◽  
Mammary Epithelial ◽  
Astragaloside Iv ◽  
Bovine Mammary Epithelial Cells

Ammonia is one of the major toxic components of metabolites in blood and tissues of high-producing dairy cows and could affect the health of bovine mammary glands. Bovine mammary epithelial cells are sensitive to oxidative stress induced by intensive cell metabolism. In our previous study, we found that ammonia could induce oxidative stress, apoptosis and inflammatory responses in bovine mammary epithelial cells. In the present study, the cytoprotective effects of astragaloside IV against ammonia in vitro were explored. The results demonstrated that pretreatment of MAC-T cells with astragaloside IV could potently suppress the increase in the level of intracellular reactive oxygen species (ROS) and the rate of cell apoptosis, inhibit the ammonia-induced inflammatory responses, and rescue the decrease of cell viability. Astragaloside IV prevented ammonia-induced endoplasmic reticulum stress. Astragaloside IV also significantly suppressed the levels of BAX, caspase 3 and p53 phosphorylation in ammonia-induced MAC-T cells. Nuclear factor erythroid 2-related factor 2(Nrf2) was essential for cytoprotective effects of astragaloside IV in MAC-T cells, as knockdown of Nrf2 dramatically abolished the prosurvival effects of astragaloside IV on treated cells. Furthermore, the PI3K/AKT and ERK/MAPK pathways were responsible for the induction of Nrf2 by astragaloside IV. In conclusion, astragaloside IV played a beneficial role against ammonia-induced damage of MAC-T cells. This provides a cue for future study to use astragaloside IV as a protective and curative agent against ammonia exposure of mammary glands in dairy cows.

scholarly journals 5-ALA Attenuates the Palmitic Acid-Induced ER Stress and Apoptosis in Bovine Mammary Epithelial Cells

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1183
Author(s):  
Mst Mamuna Sharmin ◽  
Md Aminul Islam ◽  
Itsuki Yamamoto ◽  
Shin Taniguchi ◽  
Shinichi Yonekura
Keyword(s):  
T Cells ◽  
Epithelial Cells ◽  
Palmitic Acid ◽  
Er Stress ◽  
Aminolevulinic Acid ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Protective Effects ◽  
Bovine Mammary Epithelial Cells ◽  
Induced Apoptosis

The conservation of mammary gland physiology by maintaining the maximum number of mammary epithelial cells (MECs) is of the utmost importance for the optimum amount of milk production. In a state of negative energy balance, palmitic acid (PA) reduces the number of bovine MECs. However, there is no effective strategy against PA-induced apoptosis of MECs. In the present study, 5-aminolevulinic acid (5-ALA) was established as a remedial agent against PA-induced apoptosis of MAC-T cells (an established line of bovine MECs). In PA-treated cells, the apoptosis-related genes BCL2 and BAX were down- and upregulated, respectively. The elevated expression of major genes of the unfolded protein response (UPR), such as CHOP, a proapoptotic marker (C/EBP homologous protein), reduced the viability of PA-treated MAC-T cells. In contrast, 5-ALA pretreatment increased and decreased BCL2 and BAX expression, respectively. Moreover, cleaved caspase-3 protein expression was significantly reduced in the 5-ALA-pretreated group in comparison with the PA group. The downregulation of major UPR-related genes, including CHOP, extended the viability of MAC-T cells pretreated with 5-ALA and also reduced the enhanced intensity of the PA-induced expression of phospho-protein kinase R-like ER kinase. Moreover, the enhanced expression of HO-1 (antioxidant gene heme oxygenase) by 5-ALA reduced PA-induced oxidative stress (OxS). HO-1 is not only protective against OxS but also effective against ER stress. Collectively, these findings offer new insights into the protective effects of 5-ALA against PA-induced apoptosis of bovine MECs.

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