scholarly journals The function of antimicrobial peptide MPX on the apoptosis and barrier of IPEC-J2 cells

2021 ◽  
Vol 23 (102) ◽  
pp. 125-129
Author(s):  
Xueqin Zhao

The porcine intestinal epithelial cell line IPEC-J2 cells, which were isolated from neonatal piglet mid-jejunum, the main components of the intestinal epithelium and play an important role in the intestine. Escherichia coli is an important cause of diarrhea in human and animal. E.coli infections are the leading cause of travelers' diarrhea and a major cause of diarrhea in developing nations, where it can be life-threatening among children. The aim of this study is to explore MPX against E. coli to inhibit IPEC-J2 cells apoptosis and enhance cell barrier. In this study, scanning electron microscopy results found that E. coli infection caused cell apoptosis and destroyed cell membranes of IPEC-J2 cell. MPX effectively alleviated apoptosis of IPEC-J2 cells. The laser confocal results further found that MPX prevented cell apoptosis by inhibiting caspase-3 and caspase-9 activation. In addition, it was found that MPX regulated the expression of tight junction proteins ZO-1, Occludin, and Claudin-1 in IPEC-J2 cells and is closely related to Rac1 by adding a Rac1 inhibitor. The results of adhesion and invasion experiments further found that MPX inhibited bacterial adhesion and invasion through Rac1. The above results indicated that MPX has better function in inhibiting IPEC-J2 cells apoptosis and enhancing cell barrier.

2019 ◽  
Vol 20 (3) ◽  
pp. 754 ◽  
Author(s):  
Zhihang Yuan ◽  
Zengenni Liang ◽  
Jine Yi ◽  
Xiaojun Chen ◽  
Rongfang Li ◽  
...  

Medicinal herbal plants have been commonly used for intervention in different diseases and improvement of health worldwide. Koumine, an alkaloid monomer found abundantly in Gelsemium plants, can be effectively used as an antioxidant. The purpose of this study was to evaluate the potential protective effect of koumine against hydrogen peroxide (H2O2)-induced oxidative stress and apoptosis in porcine intestinal epithelial cell line (IPEC-J2 cells). MTT assays showed that koumine significantly increased cell viability in H2O2-mediated IPEC-J2 cells. Preincubation with koumine ameliorated H2O2-medicated apoptosis by decreasing reactive oxygen species (ROS) production, and efficiently suppressed the lactate dehydrogenase (LDH) release and malondialdehyde (MDA) production. Moreover, a loss of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activities was restored to normal level in H2O2-induced IPEC-J2 cells upon koumine exposure. Furthermore, pretreatment with koumine suppressed H2O2-mediated loss of mitochondrial membrane potential, caspase-9 and caspase-3 activation, decrease of Bcl-2 expression and elevation of Bax expressions. Collectively, the results of this study indicated that koumine possesses the cytoprotective effects in IPEC-J2 cells during exposure to H2O2 by suppressing production of ROS, inhibiting the caspase-3 activity and influencing the expression of Bax and Bcl-2. Koumine could potentially serve as a protective effect against H2O2-induced apoptosis.


1992 ◽  
Vol 55 (7) ◽  
pp. 555-565 ◽  
Author(s):  
NISHA V. PADHYE ◽  
MICHAEL P. DOYLE

Escherichia coli O157:H7 is now recognized as an important human pathogen. Illnesses caused by E. coli O157:H7 infection can range from self-limited, watery diarrhea to life-threatening manifestations such as hemolytic uremic syndrome or thrombotic thrombocytopenic purpura. The mode of transmission is primarily through food; however, person-to-person transmission also has been identified in some day-care center and nursing home out-breaks. Studies to date indicate that cattle are an important reservoir of the organism. Although adhesion to intestinal epithelial cells and verotoxins are considered important virulence factors in the pathogenesis of the organism, more research is are necessary to determine the exact mechanism of pathogenicity. There is need for a rapid diagnostic test for the detection of E. coli O157:H7 in food and in clinical samples. Several useful research reagents have been developed for detecting E. coli O157:H7; however, they must be applied to a procedure that is specific, sensitive, rapid, easy to use, and commercially available so that microbiological laboratories can readily use them.


2005 ◽  
Vol 125 (3) ◽  
pp. 293-305 ◽  
Author(s):  
Peter Schierack ◽  
Marcel Nordhoff ◽  
Marion Pollmann ◽  
Karl Dietrich Weyrauch ◽  
Salah Amasheh ◽  
...  

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 202-202
Author(s):  
Hang Lu ◽  
Kolapo Ajuwon

Abstract Intestinal epithelial cells represent the frontline of host defense against invasion by enteric pathogens and uptake of potentially harmful substances. Mycotoxins are deleterious feed contaminants routinely found in cereal grains used in livestock feed. Development of strategies to eliminate mycotoxins and mitigate their harmful effects on the animal is of major importance to the livestock industry. The objective of the current study was to determine the effects of deoxynivalenol (DON), a common mycotoxin contaminant, and selected prebiotic, probiotic, and essential oil blended feed additives, on viability and integrity in porcine intestinal epithelial cell line (IPEC-J2). IPEC-J2 cells were grown on either plastic or semipermeable membranes until 100% confluence and treated with DON (12.5 uM) with or without additives (0.01 mg/mL Biolex® and 0.0001 mg/mL Encinnate®, Biomatrix Inc.) Use of DON at 12.5 uM decreased the trans-epithelial electrical resistance (TEER) by 37.5% compared to control, and led to 88.7% increase in FITC-dextran permeability after 3 days of culture, but with no negative effects on cell viability. The decrease in TEER by DON was minimized in the presence of Biolex (22.2%) and Encinnate (20.5%) relative to control. Treatment with DON also decreased the mRNA (24.0%) and protein abundance (64.4%) of claudin 3 and this was partially reversed by the additives, 11.5 and 11.0% decrease for claudin 3 mRNA and 58.7 and 56.7% decrease for the protein relative to control for Biolex and Encinnate, respectively. In summary, DON led to impaired epithelial tight junction integrity and this was partially rescued by the additives tested.


2015 ◽  
Vol 63 (1) ◽  
pp. 60-68
Author(s):  
Béla Gyetvai ◽  
Ákos Jerzsele ◽  
Erzsébet Pászti-Gere ◽  
Gábor Nagy ◽  
Péter Gálfi

Gentamicin is an aminoglycoside antibiotic widely used in combination with dimethyl sulphoxide (DMSO) in topical drug formulations. It is not known, however, whether DMSO can enhance the permeation of gentamicin through biological membranes, leading to oto- and nephrotoxic side effects. A simple and reliable high-performance liquid chromatographic (HPLC) method was applied for the quantitative determination of gentamicin collected from the apical and basolateral compartments of the porcine intestinal epithelial cell line IPEC-J2 cell monolayer using fluorometric derivatisation of the analyte with fluorenylmethyloxycarbonyl chloride (FMOC) prior to chromatographic run in the presence and absence of 1% DMSO. The lack of change in transepithelial electrical resistance (TER) demonstrated that gentamicin and 1% DMSO did not affect IPEC-J2 cell monolayer integrity via the disruption of cell membranes. Chromatographic data also ascertained that gentamicin penetration across the cell monolayer even in the presence of 1% DMSO was negligible at 6 h after the beginning of apical gentamicin administration. This study further indicates that the addition of this organic solvent does not increase the incidence of toxic effects related to gentamicin permeation.


2022 ◽  
Vol 67 (4) ◽  
pp. 1-9
Author(s):  
Shulong Zhang ◽  
Kaihua Zhu ◽  
Qi Han ◽  
Quan Wang ◽  
Bin Yang

LncRNA prostate cancer-associated transcript 1 (PCAT1) is a well-known oncogene, but the mechanisms of exosomes PCAT1 in colorectal cancer (CRC) remain largely unknown. Thus, the mechanisms of exosomes lncRNA PCAT1 were investigated. The expressions of exosomes lncRNA PCAT1 in tissues from stage 0-I and stage II-III CRC patients, and intestinal epithelial cell line FHC and two CRC cell lines, HT29 and HCT8 were measured by real-time quantitative PCR. The effects of lncRNA PCAT1 on adhesion and invasion of two CRC cell lines were investigated by cell-matrix adhesion and transwell assays. In addition, the target of PCAT1 (ZNF217) was validated using an RNA immune precipitation assay. Finally, the protein levels of MTA2, MTA3, SNAI1, and E-cadherin in normal participants, stage 0-I and stage II-III CRC patients, as well as two cell lines with stable ZNF217 knockdown were investigated by western blotting. The plasma exosomal lncRNA PCAT1 was found to be significantly increased in the CRC tissues and cell lines. In addition, lncRNA PCAT1 knockdown significantly inhibited the adhesion and invasion of HT29 and HCT8 cells. RIP assay results showed lncRNA PCAT1 could target ZNF217, and downregulation of lncRNA PCAT1 could decrease the protein expressions of ZNF217 in two CRC cells lines. Moreover, ZNF217 knockdown significantly decreased MTA2, MTA3, and SNAI1 expressions, but increased E-cadherin expressions in both CRC cells lines. Exosomal lncRNA PCAT1 can promote the adhesion and invasion of CRC cells, and PCAT1 overexpression may lead to ZNF217 upregulation that regulates EMT-related MTA2/MTA3/Snai1/E-cadherin signaling


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