Apoptosis demonstration of the renal tubules after ischemia and reperfusion by acridine orange technique

Abstract

Ischemia – reperfusion injury is a complex phenomenon that results in cell damage through a biphasic process. Ischemia initiates the injury by a decrease or complete loss of energy supply needed to maintain homeostasis. Reperfusion increases cellular damage by a variety of proposed mechanisms such as inflammatory reaction and release of oxygen free radicals. I-R resulting in both pathological cell death (necrosis) and programmed cell death (apoptosis) in human and experiments. This study performed to demonstrate the apoptosis and necrosis the renal tubules under the effect of I/R in by florescent acridine orange. Twenty five male rats were divided into six groups: control group, ischemia group (40 minute clamping of renal artery) and ischemia – reperfusion group (the removal of renal clamping after 1, 3, and 6 hours).The samples were prepared for histological technique and the sections stained with acridine orange and examined by florescent microscope. The results observed mild changes in the ischemic group but the damage of the tissue and cells occurred in the reperfusion groups reaches the maximum effect after 1 and 3 hour, the necrosis and apoptosis were observed clearly.After 6 hours reperfusion, emit orange fluorescence as evidence of nuclear fragmentation (apoptosis) in cells.We concluded that the apoptosis and necrosis occurred in the reperfusion after 40 minutes ischemia; also the acridine orange stain is effective stain to investigate the apoptosis and the cellular changes.