Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells
Abstract
:1. Introduction
2. Results
2.1. L. plantarum Lac16 Inhibited the Growth and Biofilm Formation of C. perfringens
2.2. L. plantarum Lac16 Enhanced mRNA Expression of Host Defense Peptides (HDPs) in IPEC-J2 Cells
2.3. L. plantarum Lac16 Alleviated C. perfringens Infection-Associated Lactate Dehydrogenase (LDH) Leakage
2.4. L. plantarum Lac16 Suppressed the Adhesion of C. perfringens to IPEC-J2 Cells
2.5. L. plantarum Lac16 Attenuated C. perfringens-Induced Damage to Intestinal Epithelial Barrier Function
2.6. L. plantarum Lac16 Alleviated C. perfringens-Induced Inflammatory Response
2.7. L. plantarum Lac16 Alleviated the Increase in mRNA Expression Levels of Pattern Recognition Receptors (PRRs) Induced by C. perfringens Infection
2.8. L. plantarum Lac16 Alleviated Inflammatory Response Induced by C. perfringens Infection by Nuclear Factor Kappa B (NF-κB) Signaling Pathways
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Culture Conditions
4.2. Bacteriostasis Detection of LFS to C. perfringens
4.3. Determination of Biofilm Formation of C. perfringens
4.4. Co-Culture Experiment and pH Determination of Bacterial Cultures
4.5. IPEC-J2 Cells Culture
4.6. Determination of Expression Levels of HDPs by Real-Time PCR
4.7. Cytotoxicity Assay
4.8. Determination of C. perfringens Adhesion to IPEC-J2 Cells
4.9. Cell Permeability to Fluorescein Sodium
4.10. Immunofluorescence Analysis
4.11. PAS Staining
4.12. Quantitative Real-Time PCR
4.13. Western Blot Analysis
4.14. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Gene Name | Forward Sequence (5′→3′) | Reverse Sequence (5′→3′) | Accession Number |
---|---|---|---|
β-actin | CCAGGTCATCACCATCGGCAAC | CAGCACCGTGTTGGCGTAGAG | DQ845171.1 |
pBD1 | TTCCTCCTCATGGTCCTGTT | AGGTGCCGATCTGTTTCATC | NM_213838.1 |
pBD2 | TGTCTGCCTCCTCTCTTCC | AACAGGTCCCTTCAATCCTG | AY506573.1 |
pBD3 | CCTTCTCTTTGCCTTGCTCTT | GCCACTCACAGAACAGCTACC | XM_021074698.1 |
pEP2C | ACTGCTTGTTCTCCAGAGCC | TGGCACAGATGACAAAGCCT | BK005522.1 |
Mucin 2 | GGTCATGCTGGAGCTGGACAGT | TGCCTCCTCGGGGTCGTCAC | XM_021082584.1 |
IL-1β | AGAGGGACATGGAGAAGCGA | GCCCTCTGGGTATGGCTTT | NM_001302388.2 |
IL-6 | ATCAGGAGACCTGCTTGATG | TGGTGGCTTTGTCTGGATTC | NM_001252429.1 |
IL-8 | TCCTGCTTTCTGCAGCTCTC | GGGTGGAAAGGTGTGGAATG | NM_213867.1 |
TNF-α | CTGTAGGTTGCTCCCACCTG | CCAGTAGGGCGGTTACAGAC | NM_214022.1 |
TLR-1 | GTCAGTCAGCACCGCAGTAA | CAGACAAACTGGAGGGTGGT | NM_001031775 |
TLR-2 | TCACTTGTCTAACTTATCATCCTCT | TCAGCGAAGGTGTCATTATTGC | NM_213761.1 |
TLR-4 | GCCATCGCTGCTAACATCATC | CTCATACTCAAAGATACACCATCG | NM_001113039.2 |
NOD-1 | CTGTCGTCAACACCGATCCA | CCAGTTGGTGACGCAGCTT | AB187219.1 |
NOD-2 | CCTTTTGAAGATGCTGCCTG | GATTCTCTGCCCCATCGTAG | NM_001105295.1 |
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Zhou, Y.; Wang, B.; Wang, Q.; Tang, L.; Zou, P.; Zeng, Z.; Zhang, H.; Gong, L.; Li, W. Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells. Int. J. Mol. Sci. 2021, 22, 12388. https://doi.org/10.3390/ijms222212388
Zhou Y, Wang B, Wang Q, Tang L, Zou P, Zeng Z, Zhang H, Gong L, Li W. Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells. International Journal of Molecular Sciences. 2021; 22(22):12388. https://doi.org/10.3390/ijms222212388
Chicago/Turabian StyleZhou, Yuanhao, Baikui Wang, Qi Wang, Li Tang, Peng Zou, Zihan Zeng, Huihua Zhang, Li Gong, and Weifen Li. 2021. "Protective Effects of Lactobacillus plantarum Lac16 on Clostridium perfringens Infection-Associated Injury in IPEC-J2 Cells" International Journal of Molecular Sciences 22, no. 22: 12388. https://doi.org/10.3390/ijms222212388