Eastern Kentucky University
Study 3 (Williams): Slot Blot Analysis of ³-glutamyl cysteine ethyl ester in Wistar Rats with a Moderate Traumatic Brain Injury: A Dosage Dependent Approach
Institution
Eastern Kentucky University
Faculty Advisor/ Mentor
Tanea T. Reed
Abstract
Traumatic Brain Injury (TBI) has been known to demonstrate an increase in the level of oxidative stress in proteins, which leads to irreversible secondary injury, including extensive brain damage to the individual. Glutathione is a vital intracellular antioxidant used in the neutralization of reactive oxidative species (ROS). The γ-glutamyl cysteine ethyl ester (GCEE), a glutathione mimetic, can hypothetically generate a decrease in oxidative stress in the moderate traumatic brain injury Wistar rat model. GCEE was administered intraperitoneally to Wistar rats 30 minutes post injury using a dosage dependent approach of 150 mg/kg and 200 mg/kg. Sham animals underwent craniotomy and were used to ensure that the injury was consistent. Slot blot technique was performed on all samples to determine levels of oxidized protein carbonyls, 3- Nitrotyrosine (3-NT), and 4-Hydroxynonenal (HNE) in injury and GCEE treated animals. Membranes were analyzed using Adobe Photoshop and Scion Image software and showed evidence of injury in saline treated rats, with a decreased amount of oxidation in GCEE treated animals compared to those treated with saline. Treatment with GCEE lowers overall oxidative stress making this a potential post therapeutic strategy for moderate TBI.
Study 3 (Williams): Slot Blot Analysis of ³-glutamyl cysteine ethyl ester in Wistar Rats with a Moderate Traumatic Brain Injury: A Dosage Dependent Approach
Traumatic Brain Injury (TBI) has been known to demonstrate an increase in the level of oxidative stress in proteins, which leads to irreversible secondary injury, including extensive brain damage to the individual. Glutathione is a vital intracellular antioxidant used in the neutralization of reactive oxidative species (ROS). The γ-glutamyl cysteine ethyl ester (GCEE), a glutathione mimetic, can hypothetically generate a decrease in oxidative stress in the moderate traumatic brain injury Wistar rat model. GCEE was administered intraperitoneally to Wistar rats 30 minutes post injury using a dosage dependent approach of 150 mg/kg and 200 mg/kg. Sham animals underwent craniotomy and were used to ensure that the injury was consistent. Slot blot technique was performed on all samples to determine levels of oxidized protein carbonyls, 3- Nitrotyrosine (3-NT), and 4-Hydroxynonenal (HNE) in injury and GCEE treated animals. Membranes were analyzed using Adobe Photoshop and Scion Image software and showed evidence of injury in saline treated rats, with a decreased amount of oxidation in GCEE treated animals compared to those treated with saline. Treatment with GCEE lowers overall oxidative stress making this a potential post therapeutic strategy for moderate TBI.