Objective To observe the
oxidative stress, integrity of
lysosome and
apoptosis of intestinal
epithelial cells 6 (IEC-6) after
heat stress, and explore the pathogenesis of intestinal damage caused by
heat stress.
Methods In the
heat stress groups,the
cells were incubated at 43℃ for 1 hour, then, further incubated at 37℃ and 5% CO2 for 0, 1, 3, 6 and 12 hours respectively; in the
medicine intervention group, the
cells were pretreated with the
medicine 1h before
heat stress; while in
control group, the
cells were incubated at 37℃ and 5% CO2. The amount of
reactive oxygen species (ROS) was assayed with 2', 7'-dichlorofluorescin diacetate (DCFH-DA) and dihydroethidium (DHE)
staining. The stability of
lysosome membrane was checked by AO
staining.
Apoptosis was analyzed by
flow cytometry using
annexinⅤ-
FITC/PI
staining,
CCK-8 assay was used to assess cellular viability.Results Compared with
control group,
cell viability decreased and
apoptosis increased at 1 h after
heat stress, which was the most obvious at 12h after
rewarming (P<0.05). While ROS and pale
cells increased immediately after
heat stress and the increase become the most obvious (P<0.05). The
cell viability in E-64 pretreatment group was significantly improved such as
apoptosis reduction, compared with
heat stress group (P<0.05).Conclusion
Heat stress could induce robust increase of ROS, which mediates
lysosome damage and results in
cell apoptosis, thus suggesting that ROS-
lysosome pathway may
play an important
role in intestinal
injury in
heat stress.