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X/XO or H2O2 induced IPEC-J2 cell as a new in vitro model for studying apoptosis in post-weaning piglets

文献类型： 外文期刊

作者： Cai, Xuan 1 ; Zhu, Lihui 1 ; Chen, Xiaolian 3 ; Sheng, Yongshuai 1 ; Guo, Qi 1 ; Bao, Jian 1 ; Xu, Jianxiong 1 ;

作者机构： 1.Shanghai Jiao Tong Univ, Sch Agr & Biol, Shanghai, Peoples R China

2.Shanghai Key Lab Vet & Biotechnol, Shanghai 200240, Peoples R China

3.Jiangxi Acad Agr Sci, Inst Anim Husb & Vet Sci, Nanchang 330200, Peoples R China

关键词： Apoptosis;Oxidative stress;Cell model;IPEC-J2;Weaning

期刊名称：CYTOTECHNOLOGY （ 影响因子：2.058； 五年影响因子：2.362 ）

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收录情况： SCI

摘要： We previously demonstrated that intestinal epithelial cell apoptosis in weaned piglets is much more serious than that observed in sucking piglets and is related to oxidative stress during weaning. It is difficult to study the apoptosis mechanisms only using in vivo methods because of the limit of existing research technology. An in vitro cellular system is required for piglet intestinal epithelial cell apoptosis research. In this study, a non-tumorigenic epithelial cell line, IPEC-J2 cells, was employed as a cell model. Hydrogen peroxide and xanthine/xanthine oxidase (X/XO) were both used and compared for apoptosis modeling. The concentrations of hydrogen peroxide and XO were selected and verified using cell viability analysis, the comet assay and flow cytometry. Intracellular ROS were measured using fluorescent probes. Additionally, the expression levels of the apoptosis-related genes Fas, Bcl-2, P53, Caspase 3, Caspase 8, and Caspase 9 were analyzed using quantitative RT-PCR. The results indicated the optimal modeling method is a final concentration of 0.5 mM H2O2 incubated with IPEC-J2 cells for 1 h at 37 A degrees C in 5 % CO2 for hydrogen peroxide-induced apoptosis modeling, and a final concentration of 250 mu M X/50 U/L XO incubated with IPEC-J2 cells for 6 h at 37 A degrees C in 5 % CO2 for X/XO-induced apoptosis modeling. For the apoptotic pathway, the X/XO modeling method is more similar to 21 days weaning piglets. Therefore, we suggest that X/XO modeling with IPEC-J2 cells be used as an in vitro cell culture model for weaning piglet intestinal epithelial cell apoptosis.

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