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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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Related in: MedlinePlus

Expression and quantification of selected proteins in 661W cells treated with caffeic acid phenethyl ester (CAPE) and H2O2. A: Expression and quantification of heme oxygenase 1 (HO-1), cyclooxygenase 2 (COX-2), and IκBα proteins in 661W cells was measured by western blot analysis. Proteins were extracted and subjected to western blotting with anti-HO-1, anti-COX-2, and anti-IκBα antibodies. Lane 1(NT): no treatment; lanes 2 and 3 (caffeic acid phenethyl ester [CAPE]): CAPE treated; lanes 4 and 5 (H2O2): H2O2 treated; lanes 6 and 7 (C+H): pretreated with CAPE, then with H2O2. B: Quantification of COX-2 and IκBα in 661W cells with western blotting. Quantification of COX-2 and IκBα was obtained with densitometric analysis, and normalized with β-actin. (n=3–6; *: p<0.05, by the Student t test).
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f3: Expression and quantification of selected proteins in 661W cells treated with caffeic acid phenethyl ester (CAPE) and H2O2. A: Expression and quantification of heme oxygenase 1 (HO-1), cyclooxygenase 2 (COX-2), and IκBα proteins in 661W cells was measured by western blot analysis. Proteins were extracted and subjected to western blotting with anti-HO-1, anti-COX-2, and anti-IκBα antibodies. Lane 1(NT): no treatment; lanes 2 and 3 (caffeic acid phenethyl ester [CAPE]): CAPE treated; lanes 4 and 5 (H2O2): H2O2 treated; lanes 6 and 7 (C+H): pretreated with CAPE, then with H2O2. B: Quantification of COX-2 and IκBα in 661W cells with western blotting. Quantification of COX-2 and IκBα was obtained with densitometric analysis, and normalized with β-actin. (n=3–6; *: p<0.05, by the Student t test).

Mentions: The expression of select proteins involved in cellular protective and inflammatory signaling was assayed. As shown by the gene expression studies, treatment of CAPE alone induced HO-1 protein expression to a significant level (Figure 3A), and interestingly CAPE action on HO-1 protein persisted even after 6 h of treatment with 1 mM H2O2 (Figure 3A: C+H). In addition, the level of COX-2, an inducible enzyme that acts as a dioxygenase, a peroxidase, and a potent mediator of inflammation, increased (Figure 3A). Quantification analysis showed the COX-2 protein expression increased about twofold upon treatment with CAPE (p<0.05, Figure 3B), and remained high even when treated with H2O2.


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)

Expression and quantification of selected proteins in 661W cells treated with caffeic acid phenethyl ester (CAPE) and H2O2. A: Expression and quantification of heme oxygenase 1 (HO-1), cyclooxygenase 2 (COX-2), and IκBα proteins in 661W cells was measured by western blot analysis. Proteins were extracted and subjected to western blotting with anti-HO-1, anti-COX-2, and anti-IκBα antibodies. Lane 1(NT): no treatment; lanes 2 and 3 (caffeic acid phenethyl ester [CAPE]): CAPE treated; lanes 4 and 5 (H2O2): H2O2 treated; lanes 6 and 7 (C+H): pretreated with CAPE, then with H2O2. B: Quantification of COX-2 and IκBα in 661W cells with western blotting. Quantification of COX-2 and IκBα was obtained with densitometric analysis, and normalized with β-actin. (n=3–6; *: p<0.05, by the Student t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Expression and quantification of selected proteins in 661W cells treated with caffeic acid phenethyl ester (CAPE) and H2O2. A: Expression and quantification of heme oxygenase 1 (HO-1), cyclooxygenase 2 (COX-2), and IκBα proteins in 661W cells was measured by western blot analysis. Proteins were extracted and subjected to western blotting with anti-HO-1, anti-COX-2, and anti-IκBα antibodies. Lane 1(NT): no treatment; lanes 2 and 3 (caffeic acid phenethyl ester [CAPE]): CAPE treated; lanes 4 and 5 (H2O2): H2O2 treated; lanes 6 and 7 (C+H): pretreated with CAPE, then with H2O2. B: Quantification of COX-2 and IκBα in 661W cells with western blotting. Quantification of COX-2 and IκBα was obtained with densitometric analysis, and normalized with β-actin. (n=3–6; *: p<0.05, by the Student t test).
Mentions: The expression of select proteins involved in cellular protective and inflammatory signaling was assayed. As shown by the gene expression studies, treatment of CAPE alone induced HO-1 protein expression to a significant level (Figure 3A), and interestingly CAPE action on HO-1 protein persisted even after 6 h of treatment with 1 mM H2O2 (Figure 3A: C+H). In addition, the level of COX-2, an inducible enzyme that acts as a dioxygenase, a peroxidase, and a potent mediator of inflammation, increased (Figure 3A). Quantification analysis showed the COX-2 protein expression increased about twofold upon treatment with CAPE (p<0.05, Figure 3B), and remained high even when treated with H2O2.

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