X/XO or H2O2 induced IPEC-J2 cell as a new in vitro model for studying apoptosis in post-weaning piglets
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- 作者：Xuan Cai ; Lihui Zhu ; Xiaolian Chen ; Yongshuai Sheng ; Qi Guo ; Jian Bao…
- 刊名：Cytotechnology
- 出版年：2016
- 出版时间：August 2016
- 年：2016
- 卷：68
- 期：4
- 页码：713-724
- 全文大小：1,647 KB
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Chemistry
Biotechnology
Biomedicine
Biochemistry - 出版者：Springer Netherlands
- ISSN：1573-0778
- 卷排序：68
文摘
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 °C in 5 % CO2 for hydrogen peroxide-induced apoptosis modeling, and a final concentration of 250 μM X/50 U/L XO incubated with IPEC-J2 cells for 6 h at 37 °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.KeywordsApoptosisOxidative stressCell modelIPEC-J2Weaning