Accession Number:

ADA285364

Title:

Skeletal Muscle Ischemia and Heat Shock Proteins

Descriptive Note:

Annual rept. 1 Jul 1993-30 Jun 1994

Corporate Author:

CALIFORNIA UNIV SAN DIEGO LA JOLLA

Personal Author(s):

Report Date:

1994-07-26

Pagination or Media Count:

9.0

Abstract:

Blood loss resulting in decreased organ perfusion and subsequent ischemic injury of cardiac and skeletal muscle presents a significant problem for the soldier in combat. Recent findings have indicated that different forms of noxious stress including exposure to increased temperature, noxious chemical agents, and ischemia lead to increased expression of heat shock proteins HSP which have a protective effect against injury induced by noxious stimuli. We will determine in this proposal if a rat skeletal muscle derived permanent cell line, L6 cells, expressing increased amounts of HSP70 will show protection against damaged induced by simulated ischemia. To generate L6 cells which permanently overexpress the inducible HSP70 proteins, cells will be transfected with a neomycin resistance gene and the inducible HSP70 gene. Stable lines will be selected by growing L6 cells in the presence of neomycin. Cells which have the neomycin resistance gene and the HSP70 gene incorporated into their DNA will survive. Such stably transfected L6 cell lines will then be exposed to simulated ischemia consisting of hypoxia, absence of glucose, low tonicity, and resultant ischemic 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 inducible HSP70 exerts a protective effect against ischemia mediated muscle injury. Demonstrating a protective effect of HSP70 protein will make it a useful agent to reduce ischemic muscle damage in soldiers exposed to muscle injury in combat.

Subject Categories:

  • Biochemistry
  • Medicine and Medical Research
  • Stress Physiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE