Defense Date

2007

Document Type

Thesis

Degree Name

Master of Science

Department

Physiology

First Advisor

Dr. Rakesh Kukreja

Abstract

Coronary heart disease (CHD) is one of the leading causes of death every year with nearly three-fourths of all deaths caused by the disease. The challenge scientists are facing today is discovering new drugs to protect the heart against cellular damage caused by ischemia-reperfusion injury (I-R injury). Rapamycin is one such drug that has been shown to protect the heart against ischemia-induced cellular injury. Rapamycin(sirolimus) inhibits protein synthesis through inhibition of the mammalian target ofrapamycin (mTOR). This property of rapamycin has led to its current clinical applications in drug-eluting stents and in immunosuppresive treatment to organ transplant patients. The mechanism by which this drug protects against I-R injury is currently unknown. The goal of this study is to elucidate rapamycin's cardioprotective signaling pathway. We hypothesized that upregulation of Akt occurs possibly as part of a positive feedback mechanism following the inhibition of mTOR by rapamycin. Adult male ICRmice were treated with rapamycin (0.25 mg/kg, i.p.), or volume-matched DMSO (solvent for rapamycin), or rapamycin (0.25mg/kg, i.p.) plus wortmannin (WTN, 15µg/kg, i.p.),an inhibitor of phosphatidylinositol 3-kinase, or wortmannin alone (15µg/kg, i.p.). After 30 min of stabilization, the hearts were subjected to 20 minutes of global ischemia and 30 minutes of reperfusion in Langendorff model. In a separate series of experiments mice were either injected with DMSO or rapamycin for 30 minutes, 1 hour, and 2 hours before harvesting the hearts for Western blot analysis of levels of total or phosphorylated Akt at Ser473. Our results showed that rapamycin protected the heart as observed by a reductionin infarct size from 33.8 ± 2.0% in DMSO-treated hearts to 19.3 ± 4.1% in rapamycin-treated hearts; a 43% reduction. This infarct-limiting effect was completely blocked by wortmannin (29.3 ± 4.8%). However, Western blot analysis showed no change in the level of Akt phosphorylation after administration of rapamycin. Our current resultsfurther confirmed rapamycin as a potential cardio-therapeutic drug to limit infarct size,potentially through the PI3K signaling pathway. However, the exact signaling pathway of this protection still remains elusive.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

June 2008

Included in

Physiology Commons

Share

COinS