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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

Flow cytometric analysis of verapamil on the expression of Caspase-3 in HLEC. (A) Representative cytometric profile of Caspase-3 in HLEC. The percentage of Caspase-3 was successively 2.06%, 20.08% and 2.44%, in control group, H2O2 group and verapamil group (B) Caspase-3 fluorescence intensity.
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f5-bt-22-553: Flow cytometric analysis of verapamil on the expression of Caspase-3 in HLEC. (A) Representative cytometric profile of Caspase-3 in HLEC. The percentage of Caspase-3 was successively 2.06%, 20.08% and 2.44%, in control group, H2O2 group and verapamil group (B) Caspase-3 fluorescence intensity.

Mentions: The caspase-3 positive cell of each group of HLEC was tested by flow cytometry. In the presence of H2O2, the Caspase-3 expression increased to 20.08% while that of control group was 2.06%. In the verapamil group, the caspase-3 positive cell was declined to 2.44% (Fig. 5A). The data also showed that caspase-3 fluorescence intensity of H2O2 group was higher than that of control group nearly to 4.6-fold, and verapamil effectively decreased the caspase-3 fluorescence intensity nearly to 1.4-fold of control group’ s (Fig. 5A).


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)

Flow cytometric analysis of verapamil on the expression of Caspase-3 in HLEC. (A) Representative cytometric profile of Caspase-3 in HLEC. The percentage of Caspase-3 was successively 2.06%, 20.08% and 2.44%, in control group, H2O2 group and verapamil group (B) Caspase-3 fluorescence intensity.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5-bt-22-553: Flow cytometric analysis of verapamil on the expression of Caspase-3 in HLEC. (A) Representative cytometric profile of Caspase-3 in HLEC. The percentage of Caspase-3 was successively 2.06%, 20.08% and 2.44%, in control group, H2O2 group and verapamil group (B) Caspase-3 fluorescence intensity.
Mentions: The caspase-3 positive cell of each group of HLEC was tested by flow cytometry. In the presence of H2O2, the Caspase-3 expression increased to 20.08% while that of control group was 2.06%. In the verapamil group, the caspase-3 positive cell was declined to 2.44% (Fig. 5A). The data also showed that caspase-3 fluorescence intensity of H2O2 group was higher than that of control group nearly to 4.6-fold, and verapamil effectively decreased the caspase-3 fluorescence intensity nearly to 1.4-fold of control group’ s (Fig. 5A).

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