Effects of Neutrophil Extracellular Traps on Bovine Mammary Epithelial Cells in vitro

Wei, Zhengkai; Wang, Jingjing; Wang, Yanan; Wang, Chaoqun; Liu, Xiao; Han, Zhen; Fu, Yunhe; Yang, Zhengtao

doi:10.3389/fimmu.2019.01003

Cited by 55 publications

(38 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cell bioactivity was assessed by CK18, ER, PR, and PRLR proteins. Previous studies have reported that CK18 is an epithelial cell-specific protein marker; thus, the positive images of CK18 in GMECs indicated that the cells that we obtained were, in fact, epithelial cells ( 21 ). We also detected the expression of ER, PR, and PRLR in primary GMECs, which is consistent with a previous study ( 24 ).…”

Section: Discussionsupporting

confidence: 56%

“…In this study, primary purified GMECs were taken from a Laoshan dairy goat by digesting mammary gland tissue pieces with collagenase I. This method differs from that of Shi et al ( 21 ), as they obtained MECs by cultured tissue blocks. However, their method required more than 6 days; in addition, the purity of the cells was lower than that obtained by the digestion method ( 22 , 23 ).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Establishment of a Stable β-Casein Protein-Secreted Laoshan Dairy Goat Mammary Epithelial Cell Line

Zhang

Liu

et al. 2020

Front. Vet. Sci.

View full text Add to dashboard Cite

Mammary epithelial cells are widely used as models in mastitis research and as tools for mammalian bioreactors; however, the short lifespan of these cells limits their utility. Several mammal epithelial cell line models have been established; however, the secretion capacity and the bacterial sensitivity of these lines have not been effectively evaluated. In this study, a stable immortalized goat mammary epithelial cell (GMEC) line was constructed by transfection with the SV40 gene. The monoclonal cells were then passaged through more than 50 generations after puromycin selection. The GMEC line was evaluated by reverse transcriptase polymerase chain reaction, the cell cycle, karyotype analysis, detection of apoptosis, Western blotting, and β-casein (CSN2) inducible assays. The GMEC line had a strong proliferation capacity relative to the primary GMECs. GMECs had the same karyotype as the primary cells. The GMEC lines maintained basic biological properties and had estrogen, prolactin, and progesterone receptors as same the primary cells. Additionally, the cells and the cell line could synthesize and secrete β-casein proteins. Finally, the rate of apoptosis of the transfected cells suggested that the cell line could provide a useful tool for signal research and mammary gland bioreactors.

show abstract

Section: Discussionsupporting

confidence: 56%

Section: Discussionmentioning

confidence: 99%

Establishment of a Stable β-Casein Protein-Secreted Laoshan Dairy Goat Mammary Epithelial Cell Line

Zhang

Liu

et al. 2020

Front. Vet. Sci.

View full text Add to dashboard Cite

show abstract

“…Lactate dehydrogenase (LDH) is an important marker of bacterial cytotoxicity [ 28 , 29 ]. Infection of bMECs with gram-negative bacteria, including K. pneumoniae and E. coli , increases LDH release and is accompanied by substantial cell damage [ 4 , 30 ].…”

Section: Discussionmentioning

confidence: 99%

Bacteriophages isolated from dairy farm mitigated Klebsiella pneumoniae-induced inflammation in bovine mammary epithelial cells cultured in vitro

et al. 2021

View full text Add to dashboard Cite

Background Klebsiella pneumoniae, an environmental pathogen causing mastitis in dairy cattle, is often resistant to antibiotics. K. pneumoniae was used as the host bacteria to support bacteriophage replication; 2 bacteriophages, CM8-1 and SJT-2 were isolated and considered to have therapeutic potential. In the present study, we determined the ability of these 2 bacteriophages to mitigate cytotoxicity, pathomorphological changes, inflammatory responses and apoptosis induced by K. pneumoniae (bacteriophage to K. pneumoniae MOI 1:10) in bovine mammary epithelial cells (bMECs) cultured in vitro. Results Bacteriophages reduced bacterial adhesion and invasion and cytotoxicity (lactate dehydrogenase release). Morphological changes in bMECs, including swelling, shrinkage, necrosis and hematoxylin and eosin staining of cytoplasm, were apparent 4 to 8 h after infection with K. pneumoniae, but each bacteriophage significantly suppressed damage and decreased TNF-α and IL-1β concentrations. K. pneumoniae enhanced mRNA expression of TLR4, NF-κB, TNF-α, IL-1β, IL-6, IL-8, caspase-3, caspase-9 and cyt-c in bMECs and increased apoptosis of bMECs, although these effects were mitigated by treatment with either bacteriophage for 8 h. Conclusions Bacteriophages CM8-1 and SJT-2 mitigated K. pneumoniae-induced inflammation in bMECs cultured in vitro. Therefore, the potential of these bacteriophages for treating mastitis in cows should be determined in clinical trials.

show abstract

“…Although we found that Rac1 is involved in LPS-induced mastitis, the signaling pathways in this process are still unclear. NLRP3, a component of the in ammasome, is composed of the NLRP3, ASC and caspase-1 [27,28]. Once NLRP3 in ammasome activated, it can convert the inactive pro-IL-1β to a biologically active form [29].…”

Section: Discussionmentioning

confidence: 99%

Rac1 signaling regulates LPS-induced mouse mastitis via inhibition of NLRP3, NF-κB and MAPK signaling pathways

Wang¹,

Jiang²,

Wang³

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Background Bovine mastitis characterized by mammary gland inflammatory responses, is the most frequent and costly diseases in dairy cattle. Researchers have been seeking effective treatments for this disease, but still not very successful. Ras-related C3 botulinum toxin substrate 1 (Rac1) is implicated in various cellular functions, including apoptosis, ROS production and inflammatory responses, and presents an attractive therapeutic target for many diseases. However, the effects of Rac1 signaling on mastitis remain unclear. The aim of this study is to investigate the effects of Rac1 signaling on mastitis via inhibition by Rac1 specific inhibitor NSC23766 based on a murine model of lipoplysaccharide (LPS)-induced mastitis. Methods Murine mastitis model was established by perfusion of LPS, hematoxylin-eosin (H & E) staining was employed to explore the mechanisms in mouse model, qRT- PCR analysis, Western blotting analysis. Results The results revealed that NSC23766 significantly decreased the damage of mammary gland by LPS, reduced myeloperoxidase activities, and the productions of IL-1β, IL-6, TNF-α and MCP-1 gene expression in the mammary glands with LPS perfusion. Moreover, western blot analysis showed that NSC23766 inhibited the phosphoryation of p65, IκBα, ERK, and p38, and suppressed the expression of NLRP3. Conclusion These findings suggested that administration of NSC23766 prevented the development of mastitis by inhibiting NLRP3, mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways. Accordingly, this study may provide research basis for the development of new drugs against mastitis, and NSC23766 might be a potential therapeutic drugs for mastitis.

show abstract

Effects of Neutrophil Extracellular Traps on Bovine Mammary Epithelial Cells in vitro

Cited by 55 publications

References 30 publications

Establishment of a Stable β-Casein Protein-Secreted Laoshan Dairy Goat Mammary Epithelial Cell Line

Establishment of a Stable β-Casein Protein-Secreted Laoshan Dairy Goat Mammary Epithelial Cell Line

Bacteriophages isolated from dairy farm mitigated Klebsiella pneumoniae-induced inflammation in bovine mammary epithelial cells cultured in vitro

Rac1 signaling regulates LPS-induced mouse mastitis via inhibition of NLRP3, NF-κB and MAPK signaling pathways

Contact Info

Product

Resources

About