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Protective Effects of Verapamil against H2O2-Induced Apoptosis in Human Lens Epithelial Cells.

Wang Z, Wang D, Li Y, Zhang X - Biomol Ther (Seoul) (2014)

Bottom Line: The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil.Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content.Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by H2O2.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, HE's University, Shenyang, Liaoning, 110163, P.R.China.

ABSTRACT
Verapamil is used in the treatment of hypertension, angina pectoris, and atrial fibrillation. Recently, several studies have demonstrated that verapamil increased the optic nerve head blood flow and improved the retrobulbar circulation. All these show that verapamil is potentially useful for ophthalmic treatment. Thus, the aim of this study is to investigate whether verapamil could protect human lens epithelial cell (HLEC) from oxidative stress induced by H2O2 and the cellular mechanism underlying this protective function. The viability of HLEC was determined by the MTT assay and apoptotic cell death was analyzed by Hoechst 33258 staining. Moreover, Caspase-3 expression was detected by immunocytochemistry and flow cytometry analysis. We also detected Caspase-3 mRNA expression by reverse-transcription-polymerase chain reaction and the GSH content in cell culture. The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil. Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content. Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by H2O2.

No MeSH data available.


Related in: MedlinePlus

Protective effects of verapamil on H2O2-induced damage on cultured HLEC. (A) H2O2-induced cytotoxicity inHLEC. (B) Effects of verapamil on H2O2-induced damage on cultured HLEC. Each value represents the mean ± S.E.M. obtained from four culture wells per experiment, determined in three independent experiments. **p<0.05 in comparison with control; ***p<0.01 in comparison with cells exposed to H2O2 alone.
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f1-bt-22-553: Protective effects of verapamil on H2O2-induced damage on cultured HLEC. (A) H2O2-induced cytotoxicity inHLEC. (B) Effects of verapamil on H2O2-induced damage on cultured HLEC. Each value represents the mean ± S.E.M. obtained from four culture wells per experiment, determined in three independent experiments. **p<0.05 in comparison with control; ***p<0.01 in comparison with cells exposed to H2O2 alone.

Mentions: The viability of the HLEC which induced by the different concentrations of H2O2 (0: control group, 0.1– 0.8 mM) for 24 h was detected by MTT assay. Cell viability was significantly reduced in a H2O2-concentration dependent manner (Fig.1A). Consequently, 0.2 mM H2O2 (48.8±1.1% of control cell viability) was chosen for subsequent experiments. The Cell viability was higher in the cells which were pretreatment with verapamil (viability of 79.3 ± 3.3% for verapamil concentrations of 25 μg/ml) than the cells which treated with H2O2 alone (Fig. 1B). Thus, it was possibly concluded that verapamil was effective for the protection of HLEC.


Protective Effects of Verapamil against H2O2-Induced Apoptosis in Human Lens Epithelial Cells.

Wang Z, Wang D, Li Y, Zhang X - Biomol Ther (Seoul) (2014)

Protective effects of verapamil on H2O2-induced damage on cultured HLEC. (A) H2O2-induced cytotoxicity inHLEC. (B) Effects of verapamil on H2O2-induced damage on cultured HLEC. Each value represents the mean ± S.E.M. obtained from four culture wells per experiment, determined in three independent experiments. **p<0.05 in comparison with control; ***p<0.01 in comparison with cells exposed to H2O2 alone.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256036&req=5

f1-bt-22-553: Protective effects of verapamil on H2O2-induced damage on cultured HLEC. (A) H2O2-induced cytotoxicity inHLEC. (B) Effects of verapamil on H2O2-induced damage on cultured HLEC. Each value represents the mean ± S.E.M. obtained from four culture wells per experiment, determined in three independent experiments. **p<0.05 in comparison with control; ***p<0.01 in comparison with cells exposed to H2O2 alone.
Mentions: The viability of the HLEC which induced by the different concentrations of H2O2 (0: control group, 0.1– 0.8 mM) for 24 h was detected by MTT assay. Cell viability was significantly reduced in a H2O2-concentration dependent manner (Fig.1A). Consequently, 0.2 mM H2O2 (48.8±1.1% of control cell viability) was chosen for subsequent experiments. The Cell viability was higher in the cells which were pretreatment with verapamil (viability of 79.3 ± 3.3% for verapamil concentrations of 25 μg/ml) than the cells which treated with H2O2 alone (Fig. 1B). Thus, it was possibly concluded that verapamil was effective for the protection of HLEC.

Bottom Line: The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil.Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content.Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by H2O2.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, HE's University, Shenyang, Liaoning, 110163, P.R.China.

ABSTRACT
Verapamil is used in the treatment of hypertension, angina pectoris, and atrial fibrillation. Recently, several studies have demonstrated that verapamil increased the optic nerve head blood flow and improved the retrobulbar circulation. All these show that verapamil is potentially useful for ophthalmic treatment. Thus, the aim of this study is to investigate whether verapamil could protect human lens epithelial cell (HLEC) from oxidative stress induced by H2O2 and the cellular mechanism underlying this protective function. The viability of HLEC was determined by the MTT assay and apoptotic cell death was analyzed by Hoechst 33258 staining. Moreover, Caspase-3 expression was detected by immunocytochemistry and flow cytometry analysis. We also detected Caspase-3 mRNA expression by reverse-transcription-polymerase chain reaction and the GSH content in cell culture. The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil. Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content. Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by H2O2.

No MeSH data available.


Related in: MedlinePlus