Skeletal Muscle Ischemia and Heat Shock Proteins.
Final rept. 1 Jul 93-30 Jun 96,
CALIFORNIA UNIV SAN DIEGO LA JOLLA
Pagination or Media Count:
Blood loss testing in decreased organ perfusion and subsequent ischemic injury of cardiac and skeletal muscle presents a significant problem for the soldier in combat. Recent findings indicate that different forms of noxious stress including exposure to increased temperature, noxious chemical agents, and schemia lead to increased expression of heat shock proteins hsp which have a protective effect against injury induced by noxious stimuli. We wanted to determine in this proposal if a muscle derived permanent cell line expressing increased amounts of hsp7O will show protection against damage induced by simulated ischemia. To generate cell lines which permanently overexpress the inducible hsp7O hsp7Oi proteins, cells will be transfected with a neomycin resistance gene and the hsp7Oi gene. Stable lines will be selected by growing L6 cells in the presence of neomycin. Cells which have the neomycin resistance gene and the hsp7O gene incorporated into their DNA will survive. Such stably transfected cell lines will then be exposed to simulated ischemia consisting of hypoxia, absence of glucose, low tonicity, and resultant schemic damage will be determined by quantitating cell viability measured in colony formation assays, the inhibition of protein synthesis, and the release of cytoplasmic enzymes like creatine kinase. These studies will determine if hsp7Oi exerts a protective effect against ischemia mediated muscle injury. Demonstrating a protective effect of hsp7O protein will make it a useful agent to reduce ischemic muscle damage in soldiers exposed to muscle injury in combat.
- *MUSCULOSKELETAL SYSTEM
- ARMY PERSONNEL
- DEOXYRIBONUCLEIC ACIDS
- CHEMICAL AGENTS
- TOXIC AGENTS
- WOUNDS AND INJURIES
- CREATINE PHOSPHOKINASE
- Anatomy and Physiology
- Medicine and Medical Research