Protective Effect of Heat Shock Protein 70 Against Oxidative Stresses in Human Corneal Fibroblasts.
10.3346/jkms.2004.19.4.591
- Author:
Yun Sang KIM
1
;
Jung Ah HAN
;
Tae Bum CHEONG
;
Jae Chun RYU
;
Jae Chan KIM
Author Information
1. Department of Ophthalmology, College of Medicine, Chung-Ang University, Seoul, Korea. jck50ey@kornet.net
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Heat Shock Proteins 70;
Cornea;
Fibroblasts;
Oxidative Stress;
Reactive Oxygen Species;
In-Situ Nick-End Labeling
- MeSH:
Cell Survival;
Cells, Cultured;
Cornea/*cytology;
DNA Damage;
Dose-Response Relationship, Drug;
Fibroblasts/cytology/drug effects/*metabolism;
Heat;
Heat-Shock Proteins 70/genetics/*metabolism;
Humans;
In Situ Nick-End Labeling;
Nitric Oxide/metabolism;
Nitric Oxide Donors/pharmacology;
*Oxidative Stress;
Reactive Oxygen Species/metabolism;
Research Support, Non-U.S. Gov't;
S-Nitroso-N-Acetylpenicillamine/pharmacology;
Xanthine/pharmacology;
Xanthine Oxidase/pharmacology
- From:Journal of Korean Medical Science
2004;19(4):591-597
- CountryRepublic of Korea
- Language:English
-
Abstract:
We evaluated DNA protection effect of heat shock protein (HSP) against cytotoxic effects of exogenous nitric oxide (NO) and reactive oxygen intermediate (ROI). Cultured human corneal fibroblasts were divided into 4 groups. Control (Group I) was not exposed to a sub-lethal heat treatment. Other 3 groups were exposed to 43 degrees C for 1 hr, then incubated at 37 degrees C during different duration (1, 6, 24 hr, Group II, III, IV, respectively). Expression pattern of HSP 70 was analyzed by Western blot. Cell viability was measured by MTT assay and the relationship between HSP 70 expression and DNA damage was examined by terminal deoxyribonucleotidyl transferase mediated dUTP-digoxigenin nick and labeling (TUNEL) stain and single cell gel electrophoresis. Expression pattern of HSP 70 was dependent on recovery times. Cell viability following heat treatment was significantly increased and the TUNEL positive cell number was decreased at 6 hr. In single cell gel electrophoresis, tail moments were increased in a dose-dependent manner by SNAP and X/XO. Following heat treatment, tail moments showed decreased significantly at 6 hr. These results suggest that induction of HSP 70 by sub-lethal heat treatment is closely related with cytoprotective effects against oxidative stresses in human corneal fibroblasts.
