Nitric oxide attenuates ischaemia-reperfusion (I/R) injury in the diabetic liver.
BACKGROUND
Liver ischaemia-reperfusion (I/R) occurs during resuscitation from haemorrhagic shock, hepatic transplantation and anatomic resection of the liver. This injury is associated with hepatocellular enzyme release and hepatocyte necrosis. The impact of chronic illnesses such as diabetes mellitus (DM) on hepatic I/R is unknown. This study determines the effect of DM on liver I/R using a murine model of type II DM in which the leptin receptor is defective. Preliminary studies suggest that animal models of DM have impaired endothelial nitric oxide (NO) release. Other studies suggest that NO attenuates hepatic I/R in phenotypically normal animals. We postulated that DM exacerbates hepatic I/R and that exogenous NO administration will attenuate hepatocellular injury.


METHODS
Non-diabetic and diabetic (db/db) mice were anaesthetized and underwent laparotomy with the placement of a microvascular clip on the hepatic artery and portal vein supplying the medial and left lateral lobes of the liver rendering about 70% of the liver ischaemic. Hepatic ischaemia was maintained for 45 min after which time the clip was removed and the liver segments reperfused. The abdomen was closed and the animals maintained for 5 h of reperfusion. Hepatic injury was then assessed by measuring serum alanine and aspartate transaminases (ALT, AST) spectrophotometrically. Sections of liver reperfused for 24 h were stained with haematoxylin and eosin and the percentage of hepatocyte necrosis evaluated using morphometric techniques. Other animals undergoing hepatic I/R received the NO donor (DETA 100 micro g/kg, i.v. 5 min prior to reperfusion). Time-matched, sham-operated animals served as controls. The data are expressed as mean +/- SEM and analysed by ANOVA.


RESULTS
Serum AST and ALT levels were significantly higher in db/db animals vs. non-diabetics, even in the absence of hepatic I/R (P < 0.01). Serum AST and ALT levels in db/db mice undergoing hepatic I/R were nearly five times greater than that of non-diabetic animals (P < 0.01). Histologic examination of the livers of the diabetic animals undergoing I/R demonstrated significantly greater hepatocellular necrosis (zone III; 30-40%) when compared with non-diabetic animals sustaining the same injury (zone III; 3-10%). The NO donor DETA totally prevented the increase in serum ALT and AST release associated with I/R in both the diabetic and non-diabetic mice when compared with animals not receiving this agent (P < 0.01).


CONCLUSIONS
This is the first study suggesting that DM exacerbates hepatic I/R and that NO donors will prevent this hepatocellular injury in the diabetic. Sixteen million Americans have DM. Understanding the effect of this chronic illness on the inflammatory response to injury is essential to improving clinical outcomes in these medically compromised patients.
Rogers
Hollis
H
Department of Surgery, Louisiana State University Health Sciences Center, Shreveport, 71130, USA. hroger@lsuhsc.edu
Zibari
Gazi B
GB
Roberts
Jenifer
J
Turnage
Richard
R
Lefer
David J
DJ
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D004195:Disease Models, Animal; D008099:Liver / Q000098:blood supply* / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D020030:Nitric Oxide Donors / Q000494:pharmacology / Q000627:therapeutic use*; D009603:Nitroso Compounds / Q000494:pharmacology / Q000627:therapeutic use*; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000503:physiopathology*
D020030:Nitric Oxide Donors; D009603:Nitroso Compounds; C094210:2,2'-(hydroxynitrosohydrazono)bis-ethanamine; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase