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Caffeic acid phenethyl ester protects 661W cells from H2O2-mediated cell death and enhances electroretinography response in dim-reared albino rats.

Chen H, Tran JT, Anderson RE, Mandal MN - Mol. Vis. (2012)

Bottom Line: Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, has a wide range of beneficial properties.Pretreatment of 661W cells with CAPE reduced H(2)O(2)-mediated cell death in a dose-dependent manner and induced expression of heme oxygenase-1 (Ho1).CAPE can activate the antioxidative gene expression pathway in retinal cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.

ABSTRACT

Purpose: Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, has a wide range of beneficial properties. The purpose of this study was to test the protective role of CAPE in 661W cells (in vitro) against H(2)O(2)-mediated cell death and in albino rats (in vivo) against various light conditions.

Methods: The 661W cells were pretreated with CAPE and then stressed with H(2)O(2). Cell death was measured with lactate dehydrogenase (LDH) release assay, and mRNA and proteins were analyzed. Sprague Dawley rats were raised on either a control or CAPE (0.02%) diet and exposed to various light conditions for short or long periods. Retinal histology, mRNA, protein, lipid composition, and retinal function by electroretinography (ERG) were measured at the end of feeding.

Results: Pretreatment of 661W cells with CAPE reduced H(2)O(2)-mediated cell death in a dose-dependent manner and induced expression of heme oxygenase-1 (Ho1). Albino rats fed with CAPE had greater expression of Ho1 and intercellular adhesion molecule 1 (Icam1), less expression of FOS-like antigen (Fosl) and lipoxygenase 12 (Lox12) genes in the retina, less translocation of nuclear factor kappaB protein to the nucleus, and a lower molar ratio of n-3 polyunsaturated fatty acids. Further, the ERGs of the retinas of CAPE-fed rats were significantly higher than those of the control-fed rats when raised in dim light.

Conclusions: CAPE can activate the antioxidative gene expression pathway in retinal cells in vitro and in vivo. Feeding CAPE to albino rats can enhance ERG responses and change the lipid profile in the rats' retinas.

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Gene expression in 661W cells was measured by quantitative reverse transcriptase PCR (qRT–PCR). Gene expression was analyzed with the comparative Ct value method after normalizing against the housekeeping gene, Rpl19. Expression values (±SD) are presented against fold change over no-treatment value, which was set to 1.0 (n=3 samples ×3 replication assay per sample). No treatment: no treatment with CAPE or H2O2; H2O2:treated with 1 mM of H2O2 for 6 h; caffeic acid phenethyl ester (CAPE): treated with 5 μM CAPE for 3 h; H2O2 + CAPE: treated with 5 μM CAPE for 3 h, and then treated with 1 mM of H2O2 for 6 h (*: p<0.05; **: p<0.01, by the Student t test).
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f2: Gene expression in 661W cells was measured by quantitative reverse transcriptase PCR (qRT–PCR). Gene expression was analyzed with the comparative Ct value method after normalizing against the housekeeping gene, Rpl19. Expression values (±SD) are presented against fold change over no-treatment value, which was set to 1.0 (n=3 samples ×3 replication assay per sample). No treatment: no treatment with CAPE or H2O2; H2O2:treated with 1 mM of H2O2 for 6 h; caffeic acid phenethyl ester (CAPE): treated with 5 μM CAPE for 3 h; H2O2 + CAPE: treated with 5 μM CAPE for 3 h, and then treated with 1 mM of H2O2 for 6 h (*: p<0.05; **: p<0.01, by the Student t test).

Mentions: Expression of a series of genes that includes the antioxidant pathway and survival pathway were analyzed from the CAPE-treated 661W cells by using qRT–PCR. The expression data were analyzed with the comparative Ct value method after normalizing against the housekeeping gene, 60S Ribosomal protein L19 (Rpl19). Compared to the cells with no treatment, cells pretreated with CAPE for 3 h had significantly induced expression of the heme oxygenase 1 (Ho1) gene (Figure 2, 30 fold, p<0.01). These cells also had significantly induced expression of FOS-like antigen 1 (Fosl1) by 70% (p<0.05), chemokine (C-X-C motif) ligand 1 (Cxcl1) by threefold (p<0.01), and lipoxygenase 12 (Lox12; p<0.05); however, early growth response 1 (Egr1) expression decreased significantly by twofold (p<0.01).


Caffeic acid phenethyl ester protects 661W cells from H2O2-mediated cell death and enhances electroretinography response in dim-reared albino rats.

Chen H, Tran JT, Anderson RE, Mandal MN - Mol. Vis. (2012)

Gene expression in 661W cells was measured by quantitative reverse transcriptase PCR (qRT–PCR). Gene expression was analyzed with the comparative Ct value method after normalizing against the housekeeping gene, Rpl19. Expression values (±SD) are presented against fold change over no-treatment value, which was set to 1.0 (n=3 samples ×3 replication assay per sample). No treatment: no treatment with CAPE or H2O2; H2O2:treated with 1 mM of H2O2 for 6 h; caffeic acid phenethyl ester (CAPE): treated with 5 μM CAPE for 3 h; H2O2 + CAPE: treated with 5 μM CAPE for 3 h, and then treated with 1 mM of H2O2 for 6 h (*: p<0.05; **: p<0.01, by the Student t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3369890&req=5

f2: Gene expression in 661W cells was measured by quantitative reverse transcriptase PCR (qRT–PCR). Gene expression was analyzed with the comparative Ct value method after normalizing against the housekeeping gene, Rpl19. Expression values (±SD) are presented against fold change over no-treatment value, which was set to 1.0 (n=3 samples ×3 replication assay per sample). No treatment: no treatment with CAPE or H2O2; H2O2:treated with 1 mM of H2O2 for 6 h; caffeic acid phenethyl ester (CAPE): treated with 5 μM CAPE for 3 h; H2O2 + CAPE: treated with 5 μM CAPE for 3 h, and then treated with 1 mM of H2O2 for 6 h (*: p<0.05; **: p<0.01, by the Student t test).
Mentions: Expression of a series of genes that includes the antioxidant pathway and survival pathway were analyzed from the CAPE-treated 661W cells by using qRT–PCR. The expression data were analyzed with the comparative Ct value method after normalizing against the housekeeping gene, 60S Ribosomal protein L19 (Rpl19). Compared to the cells with no treatment, cells pretreated with CAPE for 3 h had significantly induced expression of the heme oxygenase 1 (Ho1) gene (Figure 2, 30 fold, p<0.01). These cells also had significantly induced expression of FOS-like antigen 1 (Fosl1) by 70% (p<0.05), chemokine (C-X-C motif) ligand 1 (Cxcl1) by threefold (p<0.01), and lipoxygenase 12 (Lox12; p<0.05); however, early growth response 1 (Egr1) expression decreased significantly by twofold (p<0.01).

Bottom Line: Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, has a wide range of beneficial properties.Pretreatment of 661W cells with CAPE reduced H(2)O(2)-mediated cell death in a dose-dependent manner and induced expression of heme oxygenase-1 (Ho1).CAPE can activate the antioxidative gene expression pathway in retinal cells in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.

ABSTRACT

Purpose: Caffeic acid phenethyl ester (CAPE), an active component of honeybee propolis, has a wide range of beneficial properties. The purpose of this study was to test the protective role of CAPE in 661W cells (in vitro) against H(2)O(2)-mediated cell death and in albino rats (in vivo) against various light conditions.

Methods: The 661W cells were pretreated with CAPE and then stressed with H(2)O(2). Cell death was measured with lactate dehydrogenase (LDH) release assay, and mRNA and proteins were analyzed. Sprague Dawley rats were raised on either a control or CAPE (0.02%) diet and exposed to various light conditions for short or long periods. Retinal histology, mRNA, protein, lipid composition, and retinal function by electroretinography (ERG) were measured at the end of feeding.

Results: Pretreatment of 661W cells with CAPE reduced H(2)O(2)-mediated cell death in a dose-dependent manner and induced expression of heme oxygenase-1 (Ho1). Albino rats fed with CAPE had greater expression of Ho1 and intercellular adhesion molecule 1 (Icam1), less expression of FOS-like antigen (Fosl) and lipoxygenase 12 (Lox12) genes in the retina, less translocation of nuclear factor kappaB protein to the nucleus, and a lower molar ratio of n-3 polyunsaturated fatty acids. Further, the ERGs of the retinas of CAPE-fed rats were significantly higher than those of the control-fed rats when raised in dim light.

Conclusions: CAPE can activate the antioxidative gene expression pathway in retinal cells in vitro and in vivo. Feeding CAPE to albino rats can enhance ERG responses and change the lipid profile in the rats' retinas.

Show MeSH
Related in: MedlinePlus