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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

(A) HLEC were observed by fluorescence microscopy after nuclei staining with Hoechst 33258. (A) Control cells; (B) cells treated with H2O2 alone; (C) cells treated with H2O2 and verapamil. The figures are representative for three different experiments. (B) Effects of verapamil on Caspase-3 expression. The Caspase-3 expression was examined by immunocytochemistry in HLEC treated with H2O2. DAPI was used for counterstaining. The bar is 50 μm in the figures.
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f3-bt-22-553: (A) HLEC were observed by fluorescence microscopy after nuclei staining with Hoechst 33258. (A) Control cells; (B) cells treated with H2O2 alone; (C) cells treated with H2O2 and verapamil. The figures are representative for three different experiments. (B) Effects of verapamil on Caspase-3 expression. The Caspase-3 expression was examined by immunocytochemistry in HLEC treated with H2O2. DAPI was used for counterstaining. The bar is 50 μm in the figures.

Mentions: The changes of nuclear morphology were assessed by Hoechst 33258 staining after H2O2-treatment. The control HLEC nuclei had a regular and oval shape (Fig. 3A). However, when the cell was exposed to H2O2 for 24 h, nuclear condensation and fragmentation were appeared. Verapamil treatment rescued the H2O2 induced nuclear morphological change.


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)

(A) HLEC were observed by fluorescence microscopy after nuclei staining with Hoechst 33258. (A) Control cells; (B) cells treated with H2O2 alone; (C) cells treated with H2O2 and verapamil. The figures are representative for three different experiments. (B) Effects of verapamil on Caspase-3 expression. The Caspase-3 expression was examined by immunocytochemistry in HLEC treated with H2O2. DAPI was used for counterstaining. The bar is 50 μm in the figures.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-bt-22-553: (A) HLEC were observed by fluorescence microscopy after nuclei staining with Hoechst 33258. (A) Control cells; (B) cells treated with H2O2 alone; (C) cells treated with H2O2 and verapamil. The figures are representative for three different experiments. (B) Effects of verapamil on Caspase-3 expression. The Caspase-3 expression was examined by immunocytochemistry in HLEC treated with H2O2. DAPI was used for counterstaining. The bar is 50 μm in the figures.
Mentions: The changes of nuclear morphology were assessed by Hoechst 33258 staining after H2O2-treatment. The control HLEC nuclei had a regular and oval shape (Fig. 3A). However, when the cell was exposed to H2O2 for 24 h, nuclear condensation and fragmentation were appeared. Verapamil treatment rescued the H2O2 induced nuclear morphological change.

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