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
Affiliations: Department of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, UPJŠ, Košice, Slovakia | Clinic of Vascular Surgery, VÚSCH, Košice, Slovakia
Note: [] Corresponding author: Vladimíra Tomečková, PhD, Department of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, UPJŠ, Trieda SNP 1, 040 66 Košice, Slovakia. Tel./Fax: +421 55 6423849; E-mail: [email protected].
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+.
Keywords: Rat liver mitochondria, ischaemia/reperfusion of small intestine, small bowel transplantation, fluorescence excitation–emission matrix
DOI: 10.3233/SPE-2011-0545
Journal: Spectroscopy, vol. 26, no. 4-5, pp. 237-243, 2011
