The Effect of Lactoferrin and Pepsin-Treated Lactoferrin on IEC-6 Cell Damage Induced by Clostridium Difficile Toxin B. Issue 1 (July 2018)

• Record Type: Journal Article
• Authors: Otake, Kosuke; Sato, Norio; Kitaguchi, Ayako; Irahara, Takayuki; Murata, Satoru; Shiraga, Keiichiro; Ogawa, Yuichi; Fujiwara, Takahiro K.; Koike, Kaoru; Yokota, Hiroyuki
• Is Part Of: Shock. Volume 50:Issue 1(2018)
• Publication Date: 2018-07
• Subjects: Clostridium difficile infections (CDI) -- cytotoxicity -- intestinal epithelial barrier -- tight junction -- zonula occludens-1 -- CD -- Clostridium difficile -- CDI -- Clostridium difficile infections -- IEC-6 cells -- rat intestinal epithelial cells -- LF -- Lactoferrin -- PLF -- pepsin-treated lactoferrin -- TJs -- tight junctions -- ZO-1 -- zonula occludens-1; Shock -- Periodicals; Shock -- Periodicals; Choc (Pathologie) -- Périodiques; Shock; Periodicals; 616.0475
• DOI: 10.1097/SHK.0000000000000990
• Languages: English
• ISSNs: 1073-2322

Abstract

Abstract : ABSTRACT: Clostridium difficile infections (CDI) have recently increased worldwide. Some CDI progress to fulminant and recurrent CDI and are associated with high mortality and morbidity. CD produces toxins A and B, which cause intestinal mucosal damage, although toxin B exhibits greater cytotoxicity. Pepsin-treated lactoferrin (PLF) is the decomposed product of lactoferrin (LF), a multifunctional glycoprotein with anti-inflammatory properties. Here, we investigate the effects of LF and PLF in toxin B-stimulated rat intestinal epithelial (IEC-6) cells. Different toxin B concentrations were added to IEC-6 cells with or without LF or PLF. Mitochondrial function and cell cytotoxicity were assessed by measuring WST-1 and LDH levels, respectively. WST-1 levels were higher in IEC-6 cells treated with toxin B and LF or PLF than in the toxin B-only control ( P < 0.05). Compared with the toxin B-only control, LDH levels significantly decreased after toxin B and LF or PLF addition ( P < 0.05). Wound restitution measurement using microscopy demonstrated significantly greater levels of wound restitution in cells treated with toxin B and LF or PLF than in those treated with toxin B alone after 12 h ( P < 0.001). Furthermore, changes in IEC-6 cell tight junctions (TJs) were evaluated by immunofluorescence microscopy and zonula occludens-1 (ZO-1) protein expression. When LF or PLF were added to IEC-6 cells, TJ structures were maintained, and ZO-1 and occludin expression was upregulated. Taken together, these results demonstrate that LF and PLF prevent the cytotoxicity of toxin B and might have the potential to control CDI.