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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.

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Related in: MedlinePlus

Resveratrol reduced the generation of ROS. A: The cells were pretreated with 10.0 and 20.0 μΜ RES for 12 h followed by treatment with 100 μM H2O2 for 2 h. The production of ROS was examined by measuring the level of ROS production using DCF-DA by flow cytometry. The result is one representative example of three separate experiments. B: Resveratrol (20.0 μM) significantly reduced the ROS generation in HLEB-3 cells **p<0.01 versus the RES negative control. The degree of 20.0 μM RES was higher than 10.0 μM RES *p<0.05.
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f2: Resveratrol reduced the generation of ROS. A: The cells were pretreated with 10.0 and 20.0 μΜ RES for 12 h followed by treatment with 100 μM H2O2 for 2 h. The production of ROS was examined by measuring the level of ROS production using DCF-DA by flow cytometry. The result is one representative example of three separate experiments. B: Resveratrol (20.0 μM) significantly reduced the ROS generation in HLEB-3 cells **p<0.01 versus the RES negative control. The degree of 20.0 μM RES was higher than 10.0 μM RES *p<0.05.

Mentions: The ROS induced by H2O2 was examined by measuring the level of ROS production using DCF-DA. HLEB-3 cells treated with 100 μM H2O2 for 2 h resulted in the production of ROS with an approximately twofold increase compared to nontreated cells. However, the cells pretreatment with RES markedly reduced the ROS levels generated by H2O2 in HLEB-3 cells (Figure 2A). Three repeated experiments showed RES pretreatment dose-dependently inhibited ROS generation induced by H2O2, which had statistical significance (Figure 2B).


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 reduced the generation of ROS. A: The cells were pretreated with 10.0 and 20.0 μΜ RES for 12 h followed by treatment with 100 μM H2O2 for 2 h. The production of ROS was examined by measuring the level of ROS production using DCF-DA by flow cytometry. The result is one representative example of three separate experiments. B: Resveratrol (20.0 μM) significantly reduced the ROS generation in HLEB-3 cells **p<0.01 versus the RES negative control. The degree of 20.0 μM RES was higher than 10.0 μM RES *p<0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Resveratrol reduced the generation of ROS. A: The cells were pretreated with 10.0 and 20.0 μΜ RES for 12 h followed by treatment with 100 μM H2O2 for 2 h. The production of ROS was examined by measuring the level of ROS production using DCF-DA by flow cytometry. The result is one representative example of three separate experiments. B: Resveratrol (20.0 μM) significantly reduced the ROS generation in HLEB-3 cells **p<0.01 versus the RES negative control. The degree of 20.0 μM RES was higher than 10.0 μM RES *p<0.05.
Mentions: The ROS induced by H2O2 was examined by measuring the level of ROS production using DCF-DA. HLEB-3 cells treated with 100 μM H2O2 for 2 h resulted in the production of ROS with an approximately twofold increase compared to nontreated cells. However, the cells pretreatment with RES markedly reduced the ROS levels generated by H2O2 in HLEB-3 cells (Figure 2A). Three repeated experiments showed RES pretreatment dose-dependently inhibited ROS generation induced by H2O2, which had statistical significance (Figure 2B).

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