A Study On A Sum Of 40 Male Rats

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A sum of 40 male rats (8 weeks of age) will be purchased, 20 obese and 20 lean rats from Charles River Laboratories in as represented in Vendrame et al9 and Terra et al15. Male rats will be used instead of female rats to decrease the variables that may modify results due to hormonal alterations during the experiment. The rodents will be purchased from a pathogen free facility to decrease the chance of infection and fed a chow based diet during the first week of transportation to the lab. The use of experimental rodents will need to obtain approval from the University of Maine Institutional Animal Care and Use Committee9. Researchers will have appropriate training to use the Small Animals Research Facility and for humane handling and care…show more content…
10 lean rats and 10 obese rats will consume this diet for 8 weeks. Tissue Collection Modeled after Vendrame at al9, the rats will fast overnight and be anaesthetized with 95% CO2 - 5% O2 for approximately 2 minutes. A cardiac puncture will exsanguinate the specimen. The blood serum will be immediately collected, centrifuged at 3000g for 15 minutes at 4 C, and stored at -80 C17. The perivascular tissue will be extracted from the thoracic aorta and snap-frozen by liquid N2 until later use. Inflammation In order to determine inflammation within the PVAT and plasma, methods of data collection will utilize quantitative real time RT-PCR and an ELISA kit to assay lipid serum measurements9,15 as to determine cytokine and adipokine concentrations. Inflammatory markers The inflammatory markers presented below have the capability of influencing one another and are intertwined through mechanisms that influence the progression of atherosclerosis and cardiovascular disease. It is important to assay multiple chemokines and adipokines that are altered in function during times of stress to ensure the statistical and evidence based portions of experiment are reliable. The adipose tissue of obese individuals expresses increased amounts of pro-inflammatory proteins and the following will be subject to testing. Obese individuals release monocyte chemoattractant protein-1 (MCP-1)11,15, which attracts monocytes to the site of inflammation and promotes specialized M1

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