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

摘要

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 (IR _x); and, a one hour ischaemia followed by reperfusion after transplant of the small intestine (TR _x). The IR treatment was further subdivided into three groups: 1, 24 h and 30 days reperfusion-IR _1, IR _24, IR _(720), 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 TR _1 and TR _6, respectively. Once treatment had been undergone, mitochondria were isolated and all five experimental groups-IR _1, IR _(24), IR _(720), Tr _1, Tr _6-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 IR _1, IR _(24), which was indicated though an increase in the fluorescence of the reduced pyridine nucleotide NADH+H ~+.