Title

Potential Protective Effects of Caffeic Acid Phenethyl Ester (CAPE) Against H202-Induced Oxidative Stress in Cardiac H9c2 Cells Compared To Common Antioxidants

Date of Award

6-2018

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Qian Chen, PhD

Second Advisor

Susan Hingley, PhD

Third Advisor

Dianzheng Zhang, PhD

Abstract

Caffeic Acid Phenethyl Ester (CAPE) is a natural compound that has previously exhibited anti-proliferative, anti-inflammation and antioxidant activities. However, CAPE's effects have not been fully elucidated in myoblasts under oxidative stress. We compared CAPE's cytoprotective effects to several known antioxidants (caffeic acid, vitamin C, and trolox) in H9c2 cells following oxidative injury by hydrogen peroxide (H202). H9c2 cells incubated with H202 treatment ( 100-700 μM, n=4) for 24 hours dosedependently reduced cell viability (assessed by a cell counting assay). Compared to a reduction in cell viability from H202 500 μM treatment (22 ± 4% of control), H9c2 cells were significantly restored by pretreatment of CAPE at 10 μM (100 ± 25%), 20 μM (112 ± 15%) and 40 μM (109 ± 15%, n=5, p<0.05) and partiallybytrolox at 50 μM (83 ± 10%) and 100 μM (89 ± 8%, n=4, p<0.05). In contrast, pretreatment ofH9c2 cells with caffeic acid (1-80 μM, n=3) and vitamin C (1000-10,000 μM, n=3) did not restore cell viability following H202-induced injury. CAPE's cardioprotective mechanism was further investigated by measuring reactive oxygen species levels via a dichlorofluorescin diacetate assay and by evaluating heme oxygenase-1 (H0-1) expression via western blot. Increases in ROS caused by H202 500 μM (239 ± 30% of control, n=3) were significantly restored to control levels with pretreatment of CAPE dose-dependently (n=3, p<0.05). Moreover, CAPE dose-dependently increased HO-I expression (n=3), and its effects cardioprotective effects were significantly diminished using a novel H0-1 inhibitor OB- 24 (n=3, p<0.05). These results suggest CAPE has cytoprotective and antioxidant activity and thus may potentially serve as a novel pharmacological agent to mitigate oxidative stress in cardiovascular diseases.

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