Overexpression of a 60-kDa heat shock protein enhances cytoprotective function of small intestinal epithelial cells.
OBJECTIVE
With the advancement of small intestinal (double balloon and capsule) endoscopy technology, incidence of small intestinal lesion caused by nonsteroidal anti-inflammatory drugs (NSAIDs) has been known to be high. However, therapy for small intestinal mucosal lesion has not yet been developed. Previous studies have shown that heat shock proteins (HSPs) are involved in cytoprotection mediated by their function as a molecular chaperone. In this study, we examined the effect of HSP60 or HSP70 overexpression on hydrogen peroxide-induced (H2O2) or indomethacin-induced cell damage in the small intestinal epithelial cells.


METHODS
cDNA of human HSP60 or HSP70 was transfected to rat small intestinal (IEC-6) cells, and HSP60- or HSP70-overexpressing cells were cloned. IEC-6 cells transfected with vector only were used as control cells. These cells were treated with H2O2 (0-0.14mM) or indomethacin (0-2.5mM). The cell viability was determined by MTT-assay. Cell necrosis was evaluated by LDH-release assay. Further, apoptosis was evaluated by caspases-3/7 activity and TUNEL assay.


RESULTS
Cell viability after H2O2 or indomethacin treatment was significantly higher in HSP60-overexpressing cells compared with that in control cells and HSP60-overexpressing cells. Apoptotic cells were also reduced in HSP60-overexpressing.


CONCLUSIONS
These results indicate that HSP60 plays an important role in protecting small intestinal mucosal cells from H2O2-induced or indomethacin-induced cell injury. HSP70-overexpressing cells did not show anti-apoptotic ability.


CONCLUSIONS
These findings possibly suggest that function of each HSP is different in the small intestine. Therefore, for the therapy of small intestinal mucosal lesion, HSP60-induction therapy could be a new therapeutic strategy.
Department of Gastroenterology, Juntendo University School of Medicine, Japan. mootaka@juntendo.ac.jp
Makiko
M
Takada
Michiro
M
Otaka
Taiji
T
Takahashi
Yuko
Y
Izumi
Kumiko
K
Tamaki
Tomoyoshi
T
Shibuya
Naoto
N
Sakamoto
Taro
T
Osada
Sou
S
Yamamoto
Ryuichi
R
Ishida
Masaru
M
Odashima
Hideaki
H
Itoh
Sumio
S
Watanabe
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002470:Cell Survival; D018834:Chaperonin 60 / Q000378:metabolism*; D003001:Cloning, Molecular; D004847:Epithelial Cells / Q000378:metabolism*; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007421:Intestine, Small / Q000293:injuries / Q000378:metabolism*; D051381:Rats
D018834:Chaperonin 60; D018840:HSP70 Heat-Shock Proteins