Inhibition of nuclear factor-kappa B activation attenuates hydrogen peroxide-induced cytotoxicity in human lens epithelial cells.
OBJECTIVE
Hydrogen peroxide (H(2)O(2)) is the major oxidant involved in cataract formation. Lens epithelial cells have been suggested to be the first site of oxidative damage. The authors investigated the relationship between H(2)O(2)-induced cytotoxicity and activation of nuclear factor kappa B (NF-kappaB) in human lens epithelial (HLE) cells.


METHODS
HLE B-3 cells were stimulated by various concentrations of H(2)O(2) in the presence or absence of pyrrolidine dithiocarbamate (PDTC), a potent inhibitor of NF-kappaB. H(2)O(2)-induced cytotoxicity was measured by lactate dehydrogenase cytotoxicity assay. Translocation of NF-kappaB was examined by Western blot and immunocytochemistry using anti-p65 antibody.


RESULTS
H(2)O(2)-induced cytotoxicity increased in a concentration-dependent manner. PDTC treatment significantly suppressed the cytotoxicity induced by H(2)O(2). After stimulated with H(2)O(2), NF-kappaB was found translocated from cytoplasm into the nuclei. PDTC treatment also inhibited the translocation of NF-kappaB.


CONCLUSIONS
NF-kappaB signal pathway may be important in the development of H(2)O(2)-induced damage in HLE cells that is involved in cataractogenesis.
Jin
Xue-Hai
XH
Department of Ophthalmology and Visual Sciences, Hokkaido University Graduate School of Medicine, North 15, West 7, Kita-ku, Sapporo 060-8638, Japan. xhjin@med.hokudai.ac.jp
Ohgami
Kazuhiro
K
Shiratori
Kenji
K
Koyama
Yoshikazu
Y
Yoshida
Kazuhiko
K
Kase
Satoru
S
Ohno
Shigeaki
S
D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014178:Translocation, Genetic / Q000187:drug effects
D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D006861:Hydrogen Peroxide