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School of Biology and Basic Medical Sciences, Suzhou Medical College of Soochow University, Suzhou 215000, China
lingqf@suda.edu.cn
收稿:2023-05-23,
录用:2023-07-06,
网络首发:2023-08-23,
纸质出版:2024-05-01
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Heat stress induced hepatocyte apoptosis in largemouth bass
ZHAO Xuqian,MAO Wenjia,LIN Zijie,et al.Heat stress induced hepatocyte apoptosis in largemouth bass Micropterussalmoides via IRE1α/TRAF2/ASK1/JNK pathway[J].Journal of Oceanology and Limnology,2024,42(03):988-1000.
Heat stress induced hepatocyte apoptosis in largemouth bass
ZHAO Xuqian,MAO Wenjia,LIN Zijie,et al.Heat stress induced hepatocyte apoptosis in largemouth bass Micropterussalmoides via IRE1α/TRAF2/ASK1/JNK pathway[J].Journal of Oceanology and Limnology,2024,42(03):988-1000. DOI:
Heat stress (HS) has been shown to adversely affect fish livers and can lead to extensive apoptosis. To investigate the relati
onship between endoplasmic reticulum (ER) stress and HS-induced apoptosis in fish livers
we isolated and cultured primary hepatocytes of largemouth bass
Micropterus
salmoides
by trypsin method
then established an in-vitro model of liver cells under HS (35 ℃). The contents of lactic dehydrogenase (LDH) and hydrogen peroxide (H
2
O
2
) were determined to evaluate the effects of HS on hepatocyte injury and oxidative stress. RT-qPCR was performed to discover the key genes in unfolded protein response (UPR) pathways involved at different HS duration. ERS inhibitor 4-PBA and IRE1α inhibitor 4μ8C were used to further investigate the effects of HS on IRE1α apoptosis pathway in hepatocytes. Results show that HS led to significant increases in the release of LDH
the content of H
2
O
2
and the expressions of oxidative protein folding genes (
ero1α
and
pdi
) under HS
suggesting severe hepatocyte injury and oxidative stress happened in heat-stressed largemouth bass hepatocytes
.
The continuous activation of IRE1α pathway genes (
grp78
grp94
atf6
perk
eif2a
atf4
chop
ire1α
traf2
ask1
jnk1
and
jnk2
) indicated that HS led significantly to ER stress. In particular
the mRNA expression levels of ER stress-related genes (
grp78
grp94
atf6
perk
ire1α
chop
jnk1
and
jnk2
) in the high temperature (HT) +4-PBA group and the HT+4μ8C group were significantly down-regulated under HS. After 4μ8C treatment
the expression levels of apoptosis-related genes (
caspase
-
2
caspase
-
3
caspase
-
6
caspase
-
7
caspase
-
8
caspase
-
9
and
caspase
-
10
) and LDH content were significantly decreased
whereas the cell survival rate was significantly increased when given 4-PBA or 4μ8C treatment. These findings demonstrate that HS could induce liver apoptosis of largemouth bass through the IRE1α pathway
which may act as a key switch mediating liver apoptosis of largemouth bass under HS.
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