Antioxidant Activity of Whole Grapes vs Resveratrol on Protection of Citrate Synthase, Calcium ATPase, Sarcoplasmic Reticular ATPase and Choline Acetyl Transferase from Oxidation by Hydrogen Peroxide on Rabbit Urinary Bladder Muscle and Mucosa
Issue Date
2014
Authors
Francis, Johdi-Ann '14
Degree
MS in Pharmaceutical Sciences
Advisor
Levin, Robert M
Committee Members
Johnson, Arnold
Dearborn, Richard
Hass, Martha A.
Feleder, Carlos
Dearborn, Richard
Hass, Martha A.
Feleder, Carlos
Journal Title
Journal ISSN
Volume Title
Abstract
Objective: Many investigators believe that resveratrol, is the primary active ingredient responsible for grapes antioxidant properties, and as such is being advertised as an antioxidant supplement in pharmacies. The objective of this study is to compare the protective effects of a standardized grape suspension with pure resveratrol on the response of rabbit bladder tissue to the oxidative effects of H202.
Methods: Four adult male rabbits were anesthetized and the bladders excised. Six full thickness strips were obtained from each rabbit for contractile studies and the balance of the bladder separated into smooth muscle and mucosa compartments by blunt dissection. The effect of H202 in the presence and absence of the grape suspension or resveratrol on the contractile response to field stimulation was quantitated. Each tissue was homogenized and the effects of increasing concentrations of H202 in the presence and absence of grape suspension or resveratrol on the contractile response to field stimulation, citrate synthase activity, Ca2+ATPase, SERCA and ChAT were determined. In addition, two chemical tests were performed comparing the antioxidant activity of the grape suspension with resveratrol. CUPRAC test for total antioxidant activity and the ability of H2O2 to generate malondialdehyde were performed.
Results: In both chemical tests, resveratrol had approximately 10 times greater antioxidant activity than the grape suspension. Citrate synthase activity was significantly higher in the mucosa, while SERCA and ChAT were higher in the muscle. The grape suspension provided significant protection of both smooth muscle and mucosal citrate synthase activity compared to resveratrol. It also provided significant protection of the muscle ChAT activity while resveratrol lowered the protection. Ca2+ATPase and SERCA were the least sensitive of all the enzymes to the peroxide damage. For the contractile studies, the grape suspension showed significant protection at all concentration of H202, for the rate of tension rise compared to the resveratrol.
Conclusion: These data show that the grape suspension displayed greater protective effect, suggesting that the antioxidant activities is due to the combination of active components found in the grape suspension and not just one, as with dietary resveratrol.
Methods: Four adult male rabbits were anesthetized and the bladders excised. Six full thickness strips were obtained from each rabbit for contractile studies and the balance of the bladder separated into smooth muscle and mucosa compartments by blunt dissection. The effect of H202 in the presence and absence of the grape suspension or resveratrol on the contractile response to field stimulation was quantitated. Each tissue was homogenized and the effects of increasing concentrations of H202 in the presence and absence of grape suspension or resveratrol on the contractile response to field stimulation, citrate synthase activity, Ca2+ATPase, SERCA and ChAT were determined. In addition, two chemical tests were performed comparing the antioxidant activity of the grape suspension with resveratrol. CUPRAC test for total antioxidant activity and the ability of H2O2 to generate malondialdehyde were performed.
Results: In both chemical tests, resveratrol had approximately 10 times greater antioxidant activity than the grape suspension. Citrate synthase activity was significantly higher in the mucosa, while SERCA and ChAT were higher in the muscle. The grape suspension provided significant protection of both smooth muscle and mucosal citrate synthase activity compared to resveratrol. It also provided significant protection of the muscle ChAT activity while resveratrol lowered the protection. Ca2+ATPase and SERCA were the least sensitive of all the enzymes to the peroxide damage. For the contractile studies, the grape suspension showed significant protection at all concentration of H202, for the rate of tension rise compared to the resveratrol.
Conclusion: These data show that the grape suspension displayed greater protective effect, suggesting that the antioxidant activities is due to the combination of active components found in the grape suspension and not just one, as with dietary resveratrol.
Citation
Francis JA. Antioxidant activity of whole grapes vs resveratrol on protection of citrate synthase, calcium ATPase, sarcoplasmic reticular ATPase and choline acetyl transferase from oxidation by hydrogen peroxide on rabbit urinary bladder muscle and mucosa [thesis]. Ann Arbor (MI): Proquest LLC; 2014. 67 p.
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