Lactobacillus johnsonii L531 Ameliorates Escherichia coli-Induced Cell Damage via Inhibiting NLRP3 Inflammasome Activity and Promoting ATG5/ATG16L1-Mediated Autophagy in Porcine Mammary Epithelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. Porcine Mammary Epithelial Cell Culture
2.3. Bacterial Strains and Growth Conditions
2.4. Immunofluorescence
2.5. Adhesion Assay
2.6. Internalization Assay
2.7. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM)
2.8. Cell Death Assay
2.9. Real-Time Quantitative PCR
2.10. Western Blotting
2.11. Statistical Analysis
3. Results
3.1. L. johnsonii L531 Pretreatment Reduces the Adhesion of E. coli to PMECs
3.2. L. johnsonii L531 Pretreatment Reduces PMEC Damage Induced by E. coli
3.3. L. johnsonii L531 Pretreatment Ameliorates Disruption of PMEC Ultrastructure and Decreased Cell Death Induced by E. coli
3.4. L. johnsonii L531 Pretreatment Ameliorates E. coli-Induced Activation of NLRP3 Inflammasome
3.5. L. johnsonii L531 Pretreatment Suppresses E. coli-Induced Cytokine and Chemokine mRNA Expression in PMECs
3.6. L. johnsonii L531 Pretreatment Reverses the Inhibitory Effect of E. coli on Autophagy in PMECs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Primers Name | Direction a | Sequence (5′→3′) | Accession Number |
---|---|---|---|
Gapdh | F | CCAGAACATCATCCCTGCTT | NM_001206359 |
R | GTCCTCAGTGTAGCCCAGGA | ||
Il-1β | F | GGCCGCCAAGATATAACTGA | NM_214055 |
R | GGACCTCTGGGTATGGCTTTC | ||
Il-18 | F | GCTGCTGAACCGGAAGACAA | NM_213997.1 |
R | AAACACGGCTTGATGTCCCT | ||
Il-6 | F | GGGAAATGTCGAGGCTGTG | NM_214399 |
R | AGGGGTGGTGGCTTTGTCT | ||
Il-8 | F | TCCTGCTTTCTGCAGCTCTC | NM_213867 |
R | GGGTGGAAAGGTGTGGAATG | ||
Tnf-α | F | GCCCACGTTGTAGCCAATGTCAAA | NM_214022 |
R | GTTGTCTTTCAGCTTCACGCCGTT | ||
Cxcl2 | F | GGAAGTTTGTCTCAACCCCGC | NM_001001861 |
R | AGCCAGTAAGTTTCCTCCATCTC | ||
Tlr4 | F | GCCATCGCTGCTAACATCATC | NM_001113039 |
R | CTCATACTCAAAGATACACCATCGG | ||
Nlrp3 | F | GAGCCTAGGAACTCGGAGGA | NM_001256770.1 |
R | GCTCATCAAAGGCACCTTGC |
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Zou, Y.-J.; Xu, J.-J.; Wang, X.; Zhu, Y.-H.; Wu, Q.; Wang, J.-F. Lactobacillus johnsonii L531 Ameliorates Escherichia coli-Induced Cell Damage via Inhibiting NLRP3 Inflammasome Activity and Promoting ATG5/ATG16L1-Mediated Autophagy in Porcine Mammary Epithelial Cells. Vet. Sci. 2020, 7, 112. https://doi.org/10.3390/vetsci7030112
Zou Y-J, Xu J-J, Wang X, Zhu Y-H, Wu Q, Wang J-F. Lactobacillus johnsonii L531 Ameliorates Escherichia coli-Induced Cell Damage via Inhibiting NLRP3 Inflammasome Activity and Promoting ATG5/ATG16L1-Mediated Autophagy in Porcine Mammary Epithelial Cells. Veterinary Sciences. 2020; 7(3):112. https://doi.org/10.3390/vetsci7030112
Chicago/Turabian StyleZou, Yun-Jing, Jia-Jia Xu, Xue Wang, Yao-Hong Zhu, Qiong Wu, and Jiu-Feng Wang. 2020. "Lactobacillus johnsonii L531 Ameliorates Escherichia coli-Induced Cell Damage via Inhibiting NLRP3 Inflammasome Activity and Promoting ATG5/ATG16L1-Mediated Autophagy in Porcine Mammary Epithelial Cells" Veterinary Sciences 7, no. 3: 112. https://doi.org/10.3390/vetsci7030112