Methods to Protect from Skeletal Cardiovascular Insufficiency: Improving Soldier Performance in Adverse Environments
Abstract
The authors compared two methods of determining the extent of skeletal muscle injury. The first method used nitroblue tetrazolium dye (NBT) to distinguish between live and injured muscles. The muscle flap was sectioned transversely into 1 mm thick segments before staining in 0.033% dye solution in the presence of 1.9 mM NADH. The viable area stained blue/purple as the mitochondrial enzyme activity yielded a blue reaction product and the nonviable area did not stain. The areas were quantified using an automated image analysis program (Image-Pro Plus, version 5.0). Infarct size was calculated as amount of infracted area divided by the total area in percentage. Results showed that NBT dye can distinguish between live versus injured areas. The second method of quantifying skeletal muscle injury was the use of Evans Blue Dye (EBD). EBD is taken up by injured muscle cells due to permeabilization of the plasma membrane during ischemia/reperfusion. They used the EBD method to investigate the protective anti-ischemic effect of adenosine. Results showed that the adenosine A1 receptor-selective agonist CCPA (2-chloro-N-cyclopentyladenosine), when administered 2 hours before a 90-minute ischemia/6 hour or a 90-minute/24-hour reperfusion period, was able to decrease the percentage of necrosis. Data were plotted as a percentage of EBD-positive cells. The reduced infarction size of the mouse hindlimb skeletal muscle by CCPA remained significant 24 hours after reperfusion in this model. In the next series of experiments, another adenosine receptor agonist, R-phenyl-2-propyladenosine (R-PIA), was tested. Adenosine receptor agonists significantly reduced the infarction size 6 hours after reperfusion in the mouse hindlimb ischemia reperfusion model. The data establish a novel model of skeletal muscle ischemia/reperfusion injury that allows reproducible quantitation of muscle injury and show that adenosine receptor activation can exert a potent cytoprotective effect in skeletal muscle.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2005
- Accession Number
- ADA456113
Entities
People
- Bruce T. Liang
Organizations
- University of Connecticut Health Center