Therapeutic Effect of Low Doses of Acenocoumarol in the Course of Ischemia/Reperfusion-Induced Acute Pancreatitis in Rats

Abstract

Intravascular activation of coagulation is observed in acute pancreatitis and is related to the severity of this inflammation. The aim of our study was to evaluate the impact of acenocoumarol therapy on the course of acute pancreatitis induced in male rats by pancreatic ischemia followed by reperfusion. Acenocoumarol at a dose of 50, 100, or 150 µg/kg/dose was administered intragastrically once a day, starting the first dose 24 h after the initiation of pancreatic reperfusion.

Results: Histological examination showed that treatment with acenocoumarol reduces pancreatic edema, necrosis, and hemorrhages in rats with pancreatitis. Moreover, the administration of acenocoumarol decreased pancreatic inflammatory infiltration and vacuolization of pancreatic acinar cells. These findings were accompanied with a reduction in the serum activity of lipase and amylase, concentration of interleukin-1β, and plasma d-Dimer concentration. Moreover, the administration of acenocoumarol improved pancreatic blood flow and pancreatic DNA synthesis. Acenocoumarol given at a dose of 150 µg/kg/dose was the most effective in the treatment of early phase acute pancreatitis. However later, acenocoumarol given at the highest dose failed to exhibit any therapeutic effect; whereas lower doses of acenocoumarol were still effective in the treatment of acute pancreatitis.

Conclusion: Treatment with acenocoumarol accelerates the recovery of ischemia/reperfusion-induced acute pancreatitis in rats.

Keywords: ">d-dimer; acenocoumarol; acute pancreatitis; amylase; interleukin-1β; lipase; pancreatic blood flow.

Figure 1

Influence of the treatment with acenocoumarol (ACN) on the prothrombin time expressed as the international normalized ratio (INR) in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.

Figure 2

Influence of the treatment with acenocoumarol (ACN) on the total histological score of pancreatic damage in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation; ^c p < 0.05 compared to IR + ACN 100 at the same time of observation; ^d p < 0.05 compared to IR + ACN 50 at the same time of observation.

Figure 3

Representative morphological features of the pancreas observed after two-day pancreatic reperfusion in the course of ischemia/reperfusion-induced acute pancreatitis. (A) rats treated with saline; (B) rats treated with acenocoumarol given at a dose of 50 µg/kg/day; (C) rats treated with acenocoumarol given at a dose of 100 µg/kg/day; (D) rats treated with acenocoumarol given at a dose of 150 µg/kg/day. Hematoxylin-eosin counterstain. Original magnification 200×.

Figure 4

Representative morphological features of the pancreas observed after fourteen-day pancreatic reperfusion in the course of ischemia/reperfusion-induced acute pancreatitis. (A) rats treated with saline; (B) rats treated with acenocoumarol given at a dose of 50µg/kg/day; (C) rats treated with acenocoumarol given at a dose of 100 µg/kg/day; (D) rats treated with acenocoumarol given at a dose of 150 µg/kg/day. Hematoxylin-eosin counterstain. Original magnification 200×.

Figure 5

Influence of treatment with acenocoumarol (ACN) on pancreatic blood flow in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation; ^c p < 0.05 compared to IR + ACN 50 at the same time of observation; ^d p < 0.05 compared to IR + ACN 100 at the same time of observation.

Figure 6

Influence of treatment with acenocoumarol (ACN) on serum activity of amylase in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.

Figure 7

Influence of treatment with acenocoumarol (ACN) on serum activity of lipase in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.

Figure 8

Influence of treatment with acenocoumarol (ACN) on serum concentration of interleukin 1β in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.

Figure 9

Influence of treatment with acenocoumarol (ACN) on pancreatic DNA synthesis in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at a dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.

Figure 10

Influence of treatment with acenocoumarol (ACN) on plasma D-Dimer concentration in the course of ischemia/reperfusion-induced acute pancreatitis (IR). ACN was given at the dose of 50, 100, or 150 µg/kg/day (ACN 50, ACN 100, or ACN 150). Mean ± SEM. n = 8 rats in each experimental group. ^a p < 0.05 compared to control rats without induction of acute pancreatitis (C), ^b p < 0.05 compared to rats with IR without treatment with ACN at the same time of observation.