J Korean Med Sci.  2004 Aug;19(4):591-597. 10.3346/jkms.2004.19.4.591.

Protective Effect of Heat Shock Protein 70 Against Oxidative Stresses in Human Corneal Fibroblasts

Affiliations
  • 1Department of Ophthalmology, College of Medicine, Chung-Ang University, Seoul, Korea. jck50ey@kornet.net
  • 2Korea Institute of Science and Technology, Seoul, Korea.

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.

Keyword

Heat Shock Proteins 70; Cornea; Fibroblasts; Oxidative Stress; Reactive Oxygen Species; In-Situ Nick-End Labeling

MeSH Terms

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
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