Abstract
Poison of intestinal induce severe health problems in human infants and young animals due to contaminating foods and feedstuffs. With the emergence of public health concerns and high-speed diffuse of drug-opposition of bacteria, the adoption of antimicrobial peptides as potential candidates in treating pathogen infections raised up. Nature Microcin J25 (MccJ25), a class of lasso peptides separated from a fecal strain of E. coli, has been replied to display powerful antimicrobial behavior. Herein, the study was to assess the usefulness of biogenic MccJ25 in the prophylaxis of ETEC K88 infection in IPEC-J2 cells. In vitro antimicrobial activity against ETEC K88 and cytotoxicity of biogenic MccJ25 were determined first. To further understand how biogenic MccJ25 mediates its impact, ETEC K88 adhesion in cells, membrane permeability [as indicated by reduced release of lactate dehydrogenase (LDH)], transepithelial electrical resistance (TEER), barrier function, and proinflammatory cytokines levels were determined in IPEC-J2 cells after treatment with biogenic MccJ25 and challenge with ETEC K88. Biogenic MccJ25 had a minimum inhibitory concentration of 0.25 μg/mL against ETEC K88, decreased ETEC K88 adhesion in cells and did not cause cytotoxicity toward cells. Furthermore, biogenic MccJ25 protects against ETEC-induced barrier dysfunction by increasing the TEER, decreasing the LDH and promoting tight junction proteins (TJPs) by promoting the assembly of occludin and claudin-1 in the tight junction complex. Biogenic MccJ25 was further found to relieve inflammation responses through modulation of interleukine-6, IL-8 and tumor necrosis factor-α levels via inhibition of mitogen-activated protein kinase (MAPK) and nuclear factor κB activation. In summary, biogenic MccJ25 can protects against ETEC K88-induced intestinal damage and inflammatory response, recommend the hidden adoption of biogenic MccJ25 as a novel prophylactic agent to reduce pathogen infection in animals, food or humans.
Highlights
The intestinal upper protective screen is the primary platoon of ward against the incursion of cause of disease microorganisms or poison components (Farhadi et al, 2003)
When the biogenic Microcin J25 (MccJ25) at the minimal inhibitory concentration (MIC) was transferred to a well prepared on a solidified agarose mixed with enterotoxigenic Escherichia coli (ETEC) Enterotoxigenic Escherichia coli K88 (K88), a distinct zone of inhibition was observed.The time-killing kinetic curve showed that an obvious decrease in bacterial growth appeared after 20 and 30 min exposure to biogenic MccJ25, indicating that ETEC K88 was rapidly killed by biogenic MccJ25 within 0.5 h (Figure 1B)
Our findings in this study revealed that the biogenic MccJ25 showed no cytotoxicity in the intestinal porcine epithelial cells J2 (IPEC-J2) cell line based on lactate dehydrogenase (LDH) and CCK-8 assays, while it exerted strong antimicrobial activity on ETEC K88 and significantly reduced ETEC K88 adhesion in cells
Summary
The intestinal upper protective screen is the primary platoon of ward against the incursion of cause of disease microorganisms or poison components (Farhadi et al, 2003). There is increasing evidence that human infants and young animals experience a high rate of intestinal diseases caused by enterotoxigenic Escherichia coli (ETEC) acquired by ingestion of contaminated food or water (Black, 1990). This well-known ETEC can destroy epithelial barrier, increase intestinal permeability, cause inflammatory responses to human or mammals, which intensifies the systemic inflammation and damages intestinal health (Johnson et al, 2010; Wu et al, 2016; Brown et al, 2018). Due to the important statue of crosstalk among gut microbiota, intestinal barrier, and inflammatory responses in gut micro-ecology (Chen et al, 2015; Zhang et al, 2017; Fuente-Nunez et al, 2018), it is critical to understand the antibacterial effect on pathogens to fullfil the full potential and consequences of antimicrobial agents
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