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Spectroscopy
Volume 26, Issue 4-5, Pages 237-243
http://dx.doi.org/10.3233/SPE-2011-0545

A novel method for the detection of liver damage using fluorescence of hepatic mitochondria in a rat model following ischaemia/reperfusion injury of the small intestine

Vladimíra Tomečková,1,3 Miroslava Štefanišinová,1 Miroslava Bilecová-Rabajdová,1 Eliška Kriššáková,1 Marián Tomečko,2 Štefan Tóth,1 Tímea Pekárová,1 and Mária Mareková1

1Department of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, UPJŠ, Košice, Slovakia
2Clinic of Vascular Surgery, VÚSCH, Košice, Slovakia
3Department of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, UPJŠ, Trieda SNP 1, 040 66 Košice, Košice, Slovakia

Copyright © 2011 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We present a novel method of assessing damage to the liver using fluorescence analysis of hepatic mitochondria following ischaemia/reperfusion of the small intestine in a rat model. This work is of substantial importance in understanding the syndrome of multiorgan failure after ischemia/transplantation of the small intestine. Mitochondria were isolated from six sample groups that had undergone three different experimental treatments: a control group; a treatment with ischaemia followed by reperfusion of the small intestine (IRx); and, a one hour ischaemia followed by reperfusion after transplant of the small intestine (TRx). The IR treatment was further subdivided into three groups: 1, 24 h and 30 days reperfusion – IR1, IR24, IR720, respectively. Concomitantly, the TR treatment was further subdivided: one group underwent a 1 h reperfusion and another group a 6 h reperfusion following ischaemia and transplant – groups TR1 and TR6, respectively. Once treatment had been undergone, mitochondria were isolated and all five experimental groups – IR1, IR24, IR720, Tr1, Tr6 – and their emission matrices were analysed compared with that of the control group (C). Comparing fluorescence values in zone A of all experimental groups with those of the control group indicated a reduction in aromatic amino acids in the mitochondria of all experimental groups. Comparison of fluorescent zone B of experimental groups with the control group identified a lack of oxygen in samples IR1, IR24, which was indicated though an increase in the fluorescence of the reduced pyridine nucleotide NADH+H+.