Table 1.
List of abbreviations.
Table 2.
Real-time PCR primer sequences.
Fig 1.
Heat- and shake-induced stress response in rats.
(A) Heat- and shake-induced stress response in rats. (A) Stress induced temperature and weight changes (n = 6), and the level of HSP27 and 70 (n = 3) detected by RT-PCR in rats. Values represent the mean ± SD rats for each group. (B) Quantification of western blot determination of P-AKT, AKT, cleaved Casp3 and LC3; (C) mRNA expression levels of Casp12, Casp8, Casp9, Atf4, Atf6, Lcn2, Dap, Zbtb16, Herpud1, and mTOR in the rat jejunum were quantified by real-time PCR. Sections of small intestine were collected from control, 1-day, or 3-day stressed rats. P-AKT and AKT were significantly decreased after 1-day or 3-day stress. Casp12, Casp8, Casp9, Atf4, Atf6, Lcn2, Dap, Zbtb16, and Atf6 levels were significantly increased, while Herpud1 and mTOR levels were decreased. Values are expressed as a percentage of control. Data are mean ± SE, n = 3 rats for each group. *P < 0.05, **P < 0.01 compared with control; t-test.
Fig 2.
Histological changes in the small intestine following transport-associated stress.
(A) Photomicrographs of H&E-stained sections of rat jejunum from the control, 1-day, and 3-day stress groups. (B) Morphological alterations in the ultrastructure of rat jejunal epithelium following treatment in the 3-day stress group. (C) Fluorescent microscopic images of TUNEL-stained sections of rat jejunum from the control, 1-day, and 3-day stress groups. The negative control was incubated without the TUNEL reaction mixture. The positive control was incubated with micrococcal nuclease to induce DNA breaks prior to the labeling procedures.
Fig 3.
ER stress and autophagy activity in rat intestine.
(A): LC3, (B): Casp3, (C): mTOR and (D): Casp12 expression in the small intestine (jejunal villus) of rats subjected to 3 days of stress treatment or control conditions. LC3, Casp3, and Casp12 levels increased while mTOR levels decreased (brown stain; sections counterstained with hematoxylin, light purple) in response to heat treatment above control levels.
Table 3.
Fold changes in the differentially expressed genes.
Fig 4.
(A) The heat map shows differentially expressed genes in the control and 3-day stress groups. Three samples were included in each group and the gene expression profiles are shown in rows. Red and green in the heat map represent up-regulation and down-regulation relative to the control, respectively. (B) Gene interaction network map of ER stress-related molecules including mTOR, Tnf, Casp12, Bcl2ls, Slc2a4 and others using MAS 3.0 based on the database.
Fig 5.
GO term enrichment graph, construction and dynamic visualization of the gene relation network.
(A) Cellular Components; 68 differentially expressed genes were related to 39 chart records in cellular components, mainly enriched in the GO related to the cell membranes, such as the organelle membrane, organelle envelope, membrane-enclosed lumen, ER, cytoplasmic vesicle, and autophagic vacuole (B) Biological Processes; 15 genes were related to autophagy, 24 to the cellular response to stress, and 11 to the response to endoplasmic reticulum stress (C) Molecular Function. There were also 22 chart records in molecular function, mainly enriched in the GO related to protein binding and calcium ion binding.
Fig 6.
The network graph of proteins and GO, and the gene pathway network graph
(A) GO protein network graph of the biological processes using MAS 3.0. (B) Gene pathway network graph of the biological processes based on KEGG using MAS 3.0.
Fig 7.
2D view of the functional annotation clustering using DAVID software
(A) Cluster of anti-apoptosis-related GO (B) Cluster of autophagy-related GO; (C) Cluster of ER stress-related GO (D) Cluster of autophagic vacuole-related GO.
Table 4.
GO analysis of the differentially expressed genes.
Table 5.
KEGG pathway analysis of the differentially expressed genes.