Limits...
Resveratrol protects human lens epithelial cells against H2O2-induced oxidative stress by increasing catalase, SOD-1, and HO-1 expression.

Zheng Y, Liu Y, Ge J, Wang X, Liu L, Bu Z, Liu P - Mol. Vis. (2010)

Bottom Line: In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H(2)O(2) induced cell death and cell apoptosis, its role in reducing H(2)O(2) induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect.The apoptosis rate and ROS generation were detected by flow cytometric analysis.Further studies showed that RES also inhibited H(2)O(2) induced p38 and JNK phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, The First Affiliated Hospital of Harbin Medical University, Harbin, PR China.

ABSTRACT

Purpose: Oxidative damage induced by H(2)O(2) treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. Whether the effects of oxidative stress could be attenuated in cultured human lens epithelial cells by incubation with resveratrol (RES) is still unknown. In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H(2)O(2) induced cell death and cell apoptosis, its role in reducing H(2)O(2) induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect.

Methods: HLEB-3 cells, a human lens epithelial cell line, were exposed to 100 muM H(2)O(2) with or without RES pre-treatment at different concentrations for different time duration. Cell viabilities were monitored by 4-[3-[4-iodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1) assay. The apoptosis rate and ROS generation were detected by flow cytometric analysis. Expression levels of superoxide dismutases-1 (SOD-1), catalase, and heme oxygenase-1 (HO-1) proteins were measured by western-blotting analysis. p38 and c-jun N terminal kinase (JNK) activation was also evaluated by western-blotting analysis.

Results: Resveratrol clearly reduced H(2)O(2) induced cell apoptosis and ROS accumulation; protected HLEB-3 cells from H(2)O(2) induced oxidative damage, and increased the expression levels of SOD-1, catalase, and HO-1. Further studies showed that RES also inhibited H(2)O(2) induced p38 and JNK phosphorylation.

Conclusions: These findings suggested that RES protected HLEB-3 cells from H(2)O(2) induced oxidative damage, presumably by inducing three antioxidative enzymes including catalase, SOD-1, and HO-1.

Show MeSH

Related in: MedlinePlus

Resveratrol upregulated the expression of SOD-1, HO-1, and catalase. A: western blotting analysis of SOD-1, HO-1, and catalase at different time points (6, 12, 24, and 48 h) after pretreatment with 20.0 μM resveratrol. B: western blotting analysis of SOD-1, HO-1, and catalase 12 h after pretreatment with different concentration of resveratrol (2.5, 5.0, 10.0, and 20.0 μM). The expression levels of RES pretreated cells significantly increased compared with control cells (p<0.05), and the expression levels were proportional to the concentration of resveratrol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2925910&req=5

f3: Resveratrol upregulated the expression of SOD-1, HO-1, and catalase. A: western blotting analysis of SOD-1, HO-1, and catalase at different time points (6, 12, 24, and 48 h) after pretreatment with 20.0 μM resveratrol. B: western blotting analysis of SOD-1, HO-1, and catalase 12 h after pretreatment with different concentration of resveratrol (2.5, 5.0, 10.0, and 20.0 μM). The expression levels of RES pretreated cells significantly increased compared with control cells (p<0.05), and the expression levels were proportional to the concentration of resveratrol.

Mentions: Since RES can significantly reduce the ROS levels and protect against H2O2 induced cell death, we presumed that RES may induce some enzymes which can protect the cells against cytotoxicity induced by H2O2. We evaluated the expression levels of three enzymes by western-blot analysis in treated HLEB-3 cells, which are known or believed to be essential in oxidative stress protection, including superoxide dismutases (SODs), heme oxygenase (HO-1), and catalase. We detected the expression levels of these proteins at 6, 12, 24, and 48 h after the initial treatment with 20.0 μM RES. We found that as time went by, the protein levels of pretreated cells were increased compared with control cells, especially at 12 h, as Figure 3A shows. Consistent with these results, the WST-1 assay showed that pretreatment with RES for 12 h was the optimal time compared with others. Then we examined the protein levels of these enzymes at 12 h after pretreatment with 2.5, 5.0, 10.0, and 20.0 μM RES and found that the expression levels of these proteins were proportional to the concentration of RES, as Figure 3B shows. These results indicated that SOD-1, HO-1, and catalase may play an important role in the protective action of RES.


Resveratrol protects human lens epithelial cells against H2O2-induced oxidative stress by increasing catalase, SOD-1, and HO-1 expression.

Zheng Y, Liu Y, Ge J, Wang X, Liu L, Bu Z, Liu P - Mol. Vis. (2010)

Resveratrol upregulated the expression of SOD-1, HO-1, and catalase. A: western blotting analysis of SOD-1, HO-1, and catalase at different time points (6, 12, 24, and 48 h) after pretreatment with 20.0 μM resveratrol. B: western blotting analysis of SOD-1, HO-1, and catalase 12 h after pretreatment with different concentration of resveratrol (2.5, 5.0, 10.0, and 20.0 μM). The expression levels of RES pretreated cells significantly increased compared with control cells (p<0.05), and the expression levels were proportional to the concentration of resveratrol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2925910&req=5

f3: Resveratrol upregulated the expression of SOD-1, HO-1, and catalase. A: western blotting analysis of SOD-1, HO-1, and catalase at different time points (6, 12, 24, and 48 h) after pretreatment with 20.0 μM resveratrol. B: western blotting analysis of SOD-1, HO-1, and catalase 12 h after pretreatment with different concentration of resveratrol (2.5, 5.0, 10.0, and 20.0 μM). The expression levels of RES pretreated cells significantly increased compared with control cells (p<0.05), and the expression levels were proportional to the concentration of resveratrol.
Mentions: Since RES can significantly reduce the ROS levels and protect against H2O2 induced cell death, we presumed that RES may induce some enzymes which can protect the cells against cytotoxicity induced by H2O2. We evaluated the expression levels of three enzymes by western-blot analysis in treated HLEB-3 cells, which are known or believed to be essential in oxidative stress protection, including superoxide dismutases (SODs), heme oxygenase (HO-1), and catalase. We detected the expression levels of these proteins at 6, 12, 24, and 48 h after the initial treatment with 20.0 μM RES. We found that as time went by, the protein levels of pretreated cells were increased compared with control cells, especially at 12 h, as Figure 3A shows. Consistent with these results, the WST-1 assay showed that pretreatment with RES for 12 h was the optimal time compared with others. Then we examined the protein levels of these enzymes at 12 h after pretreatment with 2.5, 5.0, 10.0, and 20.0 μM RES and found that the expression levels of these proteins were proportional to the concentration of RES, as Figure 3B shows. These results indicated that SOD-1, HO-1, and catalase may play an important role in the protective action of RES.

Bottom Line: In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H(2)O(2) induced cell death and cell apoptosis, its role in reducing H(2)O(2) induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect.The apoptosis rate and ROS generation were detected by flow cytometric analysis.Further studies showed that RES also inhibited H(2)O(2) induced p38 and JNK phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, The First Affiliated Hospital of Harbin Medical University, Harbin, PR China.

ABSTRACT

Purpose: Oxidative damage induced by H(2)O(2) treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. Whether the effects of oxidative stress could be attenuated in cultured human lens epithelial cells by incubation with resveratrol (RES) is still unknown. In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H(2)O(2) induced cell death and cell apoptosis, its role in reducing H(2)O(2) induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect.

Methods: HLEB-3 cells, a human lens epithelial cell line, were exposed to 100 muM H(2)O(2) with or without RES pre-treatment at different concentrations for different time duration. Cell viabilities were monitored by 4-[3-[4-iodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1) assay. The apoptosis rate and ROS generation were detected by flow cytometric analysis. Expression levels of superoxide dismutases-1 (SOD-1), catalase, and heme oxygenase-1 (HO-1) proteins were measured by western-blotting analysis. p38 and c-jun N terminal kinase (JNK) activation was also evaluated by western-blotting analysis.

Results: Resveratrol clearly reduced H(2)O(2) induced cell apoptosis and ROS accumulation; protected HLEB-3 cells from H(2)O(2) induced oxidative damage, and increased the expression levels of SOD-1, catalase, and HO-1. Further studies showed that RES also inhibited H(2)O(2) induced p38 and JNK phosphorylation.

Conclusions: These findings suggested that RES protected HLEB-3 cells from H(2)O(2) induced oxidative damage, presumably by inducing three antioxidative enzymes including catalase, SOD-1, and HO-1.

Show MeSH
Related in: MedlinePlus