1. Functional Characterization of Porcine NK-Lysin: A Novel Immunomodulator That Regulates Intestinal Inflammatory Response.
- Author
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Lin Q, Fu Q, Chen D, Yu B, Luo Y, Huang Z, Zheng P, Mao X, Yu J, Luo J, Yan H, and He J
- Subjects
- Animals, Anti-Bacterial Agents chemistry, Anti-Bacterial Agents metabolism, Anti-Bacterial Agents pharmacology, Bacteria drug effects, Cell Line, Immunologic Factors chemistry, Immunologic Factors pharmacology, Models, Molecular, Pore Forming Cytotoxic Proteins chemistry, Pore Forming Cytotoxic Proteins pharmacology, Proteolipids chemistry, Proteolipids pharmacology, Recombinant Proteins chemistry, Recombinant Proteins metabolism, Recombinant Proteins pharmacology, Immunologic Factors metabolism, Inflammation metabolism, Intestinal Mucosa metabolism, Pore Forming Cytotoxic Proteins metabolism, Proteolipids metabolism, Swine metabolism
- Abstract
Porcine NK-Lysine (PNKL) is a new antimicrobial peptide (AMP) identified in the small intestine. In this study, PNKL protein was obtained through heterologous expression in Escherichia coli and was estimated by SDS-PAGE at 33 kDa. The antibacterial activities of PNKL were determined using various bacterial strains and showed broad-spectrum antimicrobial activity against Gram-negative and Gram-positive bacteria. Furthermore, E. coli K88-challenged IPEC-J2 cells were used to determine PNKL influences on inflammatory responses. Hemolytic assays showed that PNKL had no detrimental impact on cell viability. Interestingly, PNKL elevated the viability of IPEC-J2 cells exposure to E. coli K88. PNKL significantly decreased the cell apoptosis rate, and improved the distribution and abundance of tight junction protein ZO-1 in IPEC-J2 cells upon E. coli K88-challenge. Importantly, PNKL not only down regulated the expressions of inflammatory cytokines such as the IL-6 and TNF-α, but also down regulated the expressions of NF-κB, Caspase3, and Caspase9 in the E. coli K88-challenged cells. These results suggest a novel function of natural killer (NK)-lysin, and the anti-bacterial and anti-inflammatory properties of PNKL may allow it a potential substitute for conventionally used antibiotics or drugs.
- Published
- 2021
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