The effect of L-arginine and aprotinin on intestinal ischemia-reperfusion injury.
BACKGROUND
Intestinal ischemia/reperfusion (I/R) results in local mucosal injury, systemic injuries, and organ dysfunction. These injuries are characterized by altered microvascular and epithelial permeability and villous damage. Activation of neutrophils, platelets, and endothelial factors are known to be involved in this process. Cytokines such as TNF-alpha, IL-1, IL-6, and oxygen-derived free radicals are believed to be important pathogenic mediators. Capillary no-reflow is also known to play a role in I/R. The aim of our study was to examine the role of L-arginine, a known nitric oxide (NO) donor, and aprotinin, a protease inhibitor with multiple effects, on intestinal I/R.


METHODS
Pigs weighing 20-25 kg were used. Ischemia was established by clamping the superior mesenteric artery (SMA) at its origin and was sustained for 2 hours. Duration of reperfusion was 2 hours. The animals were divided into four groups: group A, the control group, which was submitted to I/R injury only; group B, in which L-arginine was administered at a rate of 5 mg/kg/min during ischemia and continuing throughout reperfusion; group C, in which aprotinin was administered with an initial bolus dose of 20,000 U/kg during ischemia followed by a continuous dose at 50 U/hour throughout reperfusion; and group D in which both substances were administered. In all groups TNF-alpha, IL-1, and IL-6 levels were measured using ELISA at baseline, 2 hours of ischemia, and 1 hour and 2 hours of reperfusion. SMA blood flow was measured with a Doppler probe at baseline, 10 min, 1 hour, and 2 hours of reperfusion. Histological changes of the intestinal mucosa were examined and graded on a five-point scale in all groups.


RESULTS
In the control group, levels of TNF-alpha, IL-1, and IL-6 were significantly increased during reperfusion (p < 0.05) compared to baseline. Administration of L-arginine and aprotinin led to suppression of the release of TNF-alpha, IL-1, and IL-6 during reperfusion in a statistically significant manner (all p < 0.05). A synergistic or additive effect of L-arginine and aprotinin was not observed. SMA blood flow in the control group was decreased (p > 0.05) during reperfusion compared to baseline. In animals treated with L-arginine and aprotinin, SMA blood flow during reperfusion was significantly increased (p < 0.05) compared to the control group. Histologic examination of the intestinal mucosa was characterized by flattening of the villi and necrosis in the control group. In the treated animals, less severe histological changes were noted.


CONCLUSIONS
Administration of L: -arginine and aprotinin may lead to amelioration of intestinal I/R injury. We did not note a synergistic or additive effect of these two substances. These findings warrant further studies in clinical settings for future treatment efforts.
Spanos
Constantine P
CP
Department of Surgery, Aristotelian University, Thessaloniki, Greece 54623. costasspanos@hotmail.com
Papaconstantinou
Panagiota
P
Spanos
Panagiotis
P
Karamouzis
Michael
M
Lekkas
George
G
Papaconstantinou
Christos
C
D000818:Animals; D007611:Aprotinin / Q000494:pharmacology*; D001120:Arginine / Q000494:pharmacology*; D007375:Interleukin-1 / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D007413:Intestinal Mucosa / Q000473:pathology; D007422:Intestines / Q000098:blood supply*; D008297:Male; D015427:Reperfusion Injury / Q000097:blood / Q000473:pathology / Q000503:physiopathology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013152:Splanchnic Circulation; D034421:Sus scrofa; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
D007375:Interleukin-1; D015850:Interleukin-6; D015842:Serine Proteinase Inhibitors; D014409:Tumor Necrosis Factor-alpha; D007611:Aprotinin; D001120:Arginine