Enterocyte viability and mitochondrial function after graded intestinal ischemia and reperfusion in rats.
Ischemia/reperfusion of the small intestine can lead to metabolic and structural alterations in the mucosa. Cellular dysfunction occurs when mitochondrial metabolism is compromised, which may ultimately lead to impaired organ function. The aims of this study were to assess the suppression of cellular and mitochondrial oxidative metabolism and involvement of mitochondria in the ischemia/reperfusion injury. The mitochondria were prepared from isolated enterocytes obtained from the small intestine of anesthetized adult rats following different time periods of ischemia and ischemia followed by 5 min reperfusion. Cellular and mitochondrial function were assessed using MTT (3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) reduction assay. Ischemia of increasing time periods caused a progressive decrease in cellular and mitochondrial MTT reduction in enterocytes and reperfusion showed further decrease of MTT formazan formation. Inclusion of 1 mM succinate, as respiratory substrate, showed reversal of suppression of mitochondrial function in 30-60 min ischemia whereas 90 min ischemia or short time period ischemia followed by 5 min reperfusion indicated an irreversible damage to mitochondria. This study indicated that mitochondria are a sensitive target of damage due to oxygen deficiency and possibly due to sudden burst of oxygen free radicals. Mitochondria can withstand short periods of ischemia whereas long duration ischemia or reperfusion results in irreversible damage to mitochondrial function.
Madesh
M
M
Wellcome Trust Research Laboratory, Christian Medical College Hospital, Vellore, India.
Bhaskar
L
L
Balasubramanian
K A
KA
D000818:Animals; D005562:Formazans / Q000032:analysis / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology*; D007421:Intestine, Small / Q000098:blood supply*; D007511:Ischemia / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D015424:Reperfusion; D013386:Succinates / Q000378:metabolism / Q000494:pharmacology; D019802:Succinic Acid; D013778:Tetrazolium Salts / Q000032:analysis / Q000378:metabolism; D013997:Time Factors
D005562:Formazans; D013386:Succinates; D013778:Tetrazolium Salts; C079782:MTT formazan; D019802:Succinic Acid