[논문]Probiotics Inhibit Lipopolysaccharide-Induced Interleukin-8 Secretion from Intestinal Epithelial Cells

Oh, Hyun-Wook; Jeun, Gi-Hoon; Lee, Jin; Chun, Tae-Hoon; Kim, Sae-Hun

doi:10.5851/kosfa.2012.32.4.434

Probiotics Inhibit Lipopolysaccharide-Induced Interleukin-8 Secretion from Intestinal Epithelial Cells 원문보기

Korean journal for food science of animal resources = 한국축산식품학회지, v.32 no.4, 2012년, pp.434 - 440

Oh, Hyun-Wook (Division of Food Bioscience and Technology, Korea University) , Jeun, Gi-Hoon (Division of Food Bioscience and Technology, Korea University) , Lee, Jin (Division of Food Bioscience and Technology, Korea University) , Chun, Tae-Hoon (Division of Biotechnology, Korea University) , Kim, Sae-Hun (Division of Food Bioscience and Technology, Korea University)

Abstract ▼ AI-Helper

It has been suggested that probiotics could be useful for the prevention of symptomatic relapse in patients with inflammatory bowel disease (IBD). Interleukin (IL)-8 has been well recognized as one of the pro-inflammatory cytokines that could trigger inflammation and epithelial barrier dysfunction. In this study, the anti-inflammatory effects of probiotics were investigated using a human epithelial cell line (HT-29). Probiotics from infant feces and kimchi were tested for their cytotoxicity and effects on adhesion to epithelial cells. The present results show that seven strains could form 70 % adhesion on HT-29. The probiotics used in this study did not affect HT-29 cell viability. To screen anti-inflammatory lactic acid bacteria, HT-29 cells were pretreated with live and heat-killed probiotics, and lipopolysaccharide (LPS) ($1{\mu}g/mL$) was then added to stimulate the cells. The cell culture supernatant was then used to measure IL-8 secretion by ELISA, and the cell pellet was used to determine IL-8 and toll-like receptor (TLR-4) mRNA expression levels by RT-PCR. Some probiotics (KJP421, KDK411, SRK414, E4191, KY21, and KY210) exhibited anti-inflammatory effects through the repression of IL-8 secretion from HT-29 cells. In particular, Lactobacillus salivarius E4191, originating from Egyptian infant feces, not only decreased IL-8 mRNA expression, but also decreased TLR-4 expression. These results indicate that Lactobacillus salivarius E4191 may have a protective effect in intestinal epithelial cells.

주제어

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문제 정의

This study attempted to establish appropriate conditions for heat killed probiotics with immunomodulatory functions. Before measuring IL-8 secretion from HT-29, any possible toxicity of both the live and heat-killed probiotics was investigated.

제안 방법

The aim of study was to screen anti-inflammatory probiotic strains by measuring IL-8 production in lipopolysaccharide (LPS)-stimulated human intestinal epithelial HT-29 cells, to develop a combination of these strains that has synergetic effects on the inflamed model cells, and confirm the findings by mechanism analysis.

대상 데이터

A human epithelial cell (HT-29) line was obtained from the Korea Cell Line Bank (KCLB, Korea). The cells were routinely cultured at 37℃ and 5% CO₂ in RPMI 1640 medium (HyClone) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (HyClone) and 1% penicillin/streptomycin (HyClone).
The probiotic strains used in this study were obtained from the Food Microbiology Laboratory at Korea University (Seoul, Korea). The strains were originally isolated from Korean infant feces, Egyptian infant feces, and kimchi, that are listed in Table 1.

데이터처리

Differences between groups were assessed for statistical significance using the Student’s t test.

성능/효과

Lactobacillus rhamnosus GG was used as positive control. Seven of the probiotic strains (Leuconostoc paramesenteroides KJP421, Lactobacillus salivarius E4191, Leuconostoc mesenteroides subsp. mesenteroides KFM402, Leuconostoc mesenteroides subsp. mesenteroides KDK411, Leuconostoc mesenteroides subsp. mesenteroides SRK414, Lactobacillus brevis KY21, and Lactobacillus plantarum KY210) were selected for their strong interaction with HT-29 cells, with adhesion values of greater than 70% of the inoculated bacteria. In contrast, Lactobacillus salivarius SWW308 showed the lowest adhesion capacity at 45.
mesenteroides SRK 414, Lactobacillus brevis KY21, and Lactobacillus plantarum KY210 reduced the secretion of IL-8 by 71.6% (587.3±17.3 pg/mL), 71% (582.3±37.9 pg/mL), 69.2% (567.2±32.6 pg/mL), 70.6% (579±2.2 pg/mL), 63.7% (522.6±12.4 pg/mL), and 70.4% (577.6±20 pg/mL), respectively.

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