The Protective Effect of Induced Heat Shock Protein in Human Corneal Epithelial Cells

PURPOSE: The purposes of study were to assess the expression patterns of heat shock protein (HSP) after glutamine and glutamine with non- lethal heat shock treatment, to evaluate the protective effects of heat shock protein from apoptosis in cultured human corneal epithelial cell. METHODS: The cultured human corneal epithelial cells were divided into two group. One group was treated with 0, 10, 20, 30, 40, 50 mM of glutamine and the other group was exposed to 43 degrees C (heat shock) for 30 minutes with same concentration of glutamine. After glutamine and heat treatment, the expression patterns of Hsp 27, 70 were examined by western blot and immunohistochemistry. Apoptosis was induced with 80uM of etoposide. The viability (cell protection rate of heat shock protein) against apoptosis after etoposide treatment was measured by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. RESULTS: Expression of Heat shock protein 70 was not significantly effected in only glutamine treatment, but was remarkably increased in heat shock with glutamine treatment group. The increased cell number (viability= antiapoptotic effect of heat shock protein)of glutamine with heat shock group after etoposide treatment suggested that Hsp 70 appeared to be a major role in protection of Human corneal epithelial cell from apoptosis. The expression of Heat shock protein 27 was not effected in only glutamine and heat with glutamine treatment group. CONCLUSIONS: These data suggest that induced heat shock protein protect etoposide-generated apoptosis in human corneal epithelial cell.

The Effect of Induced Heat Shock Protein 33 in Human Corneal Epithelial Cell

PURPOSE: The purposes of study was to assess the expression patterns of heat shock protein 33 (HSP33) after gene transfection and to evaluate the protective effects of heat shock protein 33 from apoptosis and oxidative stress in transfected human corneal epithelial cell. METHODS: The cultured human corneal epithelial cells were divided control and experimental group. Experimental group was transfected with HSP 33. After transfection, the experimental group was treated with etoposide (induced apoptosis) and hydrogen peroxide (induced oxidative stress). The expression patterns of HSP 33 was0 examined by western blot. The viability (cell protection rate of heat shock protein) and protective effect against apoptosis after etoposide and hydrogen peroxide treatment were measured using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. RESULTS: Expression of Heat shock protein 33 was increased in the transfection group compared with non-transfection group. The increased cell number (viability=anti-apoptotic effect of heat shock protein) of transfection group with induced HSP 33 etoposide and hydrogen peroxide treatment show that Hsp 33 appeared to have protective effect in Human corneal epithelial cell from apoptosis and oxidative stress. CONCLUSIONS: These data suggest that experimentally induced heat shock protein 33 could protect etoposide-generated apoptosis and hydreogen peroxide generated oxidative stress in human corneal epithelial cell.