Adhesion and inflammatory effect of Enterococcus faecalis to intestinal porcine epithelial cells
Journal Title: Journal of Henan Agricultural University - Year 2023, Vol 57, Issue 4
Abstract
[Objective] This study was conducted to explore the pathogenicity of swine Enterococcus faecalis by observing its adhesion and inflammatory effect to enterocytes in vitro. [Method] Two swineoriginated enterococcal isolates N41 and N8 were selected and their adhesion to artificially infected IPEC-J2 cell cultures were observed by means of microscopy after Giemsa's staining, Gram's staining, indirect immunofluorescence and field emission scanning electron microscopy as well. Meanwhile, time-line expression changes of several pro-and anti-inflammatory cytokines in the infected IPEC-J2 cells were measured through fluorogenic quantitative PCR examination. The qPCR method was also applied for evaluating the expression of epithelial-mesenchymal transition markers such as E-cadherin, N-cadherin, β-catenin, snail and vimen to explore and compare the effects of Enterococcus faecalis as a conditional pathogen on cell surface marker proteins. [Result] The staining results showed that the E. faecalis bacteria adhered to the IPEC-J2 cells at the cell surface and the intercellular tight junctions, and the bacteria were rather aggregated than evenly distributed. The fluorogenic quantitative PCR data indicated that after infected by the enterococci, the IPEC-J2 cells presented similar responses in the expression of pro-inflammatory cytokines IL-1β, IL-6 and IL-8, elevating at 1 h to 4 h PI(post infection), and decreasing at 4h to 6h PI yet still overcasting those of the control group. Contrarily, down-regulated expression was seen in the anti-inflammatory IL-12 and TGF-βlevels, especially that of IL-12. It was also noticed that there were differences between the two isolates in regulating the cytokine transcription of the infected enterocytes: the N41 bacteria being more potent in upregulating the pro-inflammatory cytokine levels while the N8 in down-regulating the anti-inflammatory responses. The detection results of EMT markers showed that, compared with the pathogenic E. coli O157:H7, E. faecalis presented less influence on the expression levels of EMT markers. [Conclusion] It is confirmed that swine E. faecalis possesses the in vitro invasion and inflammatory potentials.
Authors and Affiliations
Yue JIN, Binzhou LI, Zhenzhen GUO, Yabin WANG, Zhanyong WEI, Liying CHEN
Keywords
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- EP ID EP769233
- DOI 10.16445/j.cnki.1000-2340.20230625.001
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