Abstract
Intestinal cells can sense the presence of pathogens and trigger many important signaling pathways to maintain tissue homeostasis and normal function. Escherichia coli and lipopolysaccharides (LPS) are the main pathogenic factors of intestinal disease in pigs. However, the roles of endoplasmic reticulum stress (ERS) and its mediated apoptosis in intestinal malfunction induced by E. coli or LPS remain unclear. In the present study, we aimed to evaluate whether ERS could be activated by E. coli fed to piglets and whether the underlying mechanisms of this disease process could be exploited. Eighteen weaned pigs (21 days old) were randomly assigned to one of two treatment groups (n = 9 per group). After pre-feeding for 1 week, the diets of the piglets in one group were supplemented with E. coli (W25 K, 109 cells kg−1 diet) for 7 days. At the end of the experiment, all piglets were slaughtered to collect jejunum and ileum samples. Western blotting and immunofluorescence experiments were used to determine the expression levels and histological locations of ERS and its downstream signaling proteins. The intestinal porcine epithelial cell line J2 (IPEC-J2) was used as in vitro model to investigate the possible mechanism. The results showed that E. coli supplementation in the diet increased the GRP78 expression in the jejunum and ileum, especially in the jejunal epithelium and ileac germinal center, and elevated the expression levels of CHOP (in both the jejunum and ileum) and caspase-11 (in the ileum), indicating that ERS and CHOP–caspase-11 dependent apoptosis were activated in the porcine small intestine. Moreover, as demonstrated by in vitro experiments, the CHOP inhibitor 4-phenylbutyrate alleviated the damage to IPEC-J2 cells induced by LPS derived from E. coli. Taken together, these data strongly suggest that ERS can be triggered in the small intestine by dietary supplementation with E. coli and that CHOP–caspase-11 dependent apoptosis may play a key role in maintaining normal homeostasis of the intestine in response to pathogenic factors.
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Abbreviations
- LPS:
-
Lipopolysaccharide
- ERS:
-
Endoplasmic reticulum stress
- CHOP:
-
C/EBP homologous protein
- GRP78:
-
Glucose-regulated protein78
- E. coli :
-
Escherichia coli
- IPEC-J2:
-
Intestinal porcine epithelial cell line J2
- 4-PBA:
-
4-phenylbutyrate
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Q.J. G.L. and K.Y. conceived the experiment(s), Q.J. S.C. W.R. and G.L. conducted the experiments, Q.J. G.W. and Y.Y. analyzed the results. Q.J. G.L. and K.Y. prepared the manuscript. All authors reviewed the manuscript.
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The authors declare that they have no conflict of interest.
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The protocol for this study was approved by the Committee on the Ethics of Animal Experiments of Institute of Subtropical Agriculture, Chinese Academy of Sciences (Permit Number: 201509-03) and it was conducted out in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of Institute of Subtropical Agriculture, Chinese Academy of Sciences.
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This study was jointly supported by the National Science Foundation for Distinguished Young Scholars of Hunan Province (2016JJ1015), Key Programs of frontier scientific research of the Chinese Academy of Sciences, the National Basic Research Program of China (2013CB127306), the Open Foundation of Key Laboratory of Agro-ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences (ISA2016101), the Chinese Academy of Sciences “Hundred Talent” award (Y451022111) and the National Natural Science Foundation Project (31772642, 31472107).
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Jiang, Q., Chen, S., Ren, W. et al. Escherichia coli aggravates endoplasmic reticulum stress and triggers CHOP-dependent apoptosis in weaned pigs. Amino Acids 49, 2073–2082 (2017). https://doi.org/10.1007/s00726-017-2492-4
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DOI: https://doi.org/10.1007/s00726-017-2492-4