ABSTRACT
Title
CR3294, a novel benzamidine derivative, reduces acute pancreatitis development in mice, by inhibition of iNOS expression
Authors
M. Galuppo.
Doctorate School in Experimental Medicine – Clinical and Experimental Department of Medicine and Pharmacology – University of Messina, Italy
Doctorate School in Experimental Medicine – Clinical and Experimental Department of Medicine and Pharmacology – University of Messina, Italy
Abstract
It is generally agreed that the clou event of the pro-inflammatory cascade is the activation of gene transcription factors, which trigger the genesis of an acute or, to long-term, chronic inflammatory status, and following which the generation of potent reactive oxygen species (ROS) can lead to severe cell damage.
CR3294, a novel benzamidine derivative, is an inhibitor of pro-inflammatory gene expression with potent ROS-generation preventing effect. In vitro CR3294 prevents ROS formation and is a potent inhibitor of cytokine production. In vivo, CR3294 has been shown to reduce several types of inflammation (general and intestinal inflammation in animal models [1]), which are related to neutrophil cell influx; it reduces also some septic shock markers, such as iNOS overproduction or TNF release.
The purpose of this study is to analyze the anti-oxidant effect of CR3294 in an experimental animal model of acute pancreatitis.
Pancreatitis induced by cerulein-administration (50 μg/kg, ip) causes histological damage, accompanied by high serum levels of digestive enzymes, high cytokine production, increased expression of adhesion molecules, neutrophil infiltration, release of pro-inflammatory mediators, as well as mitochondrial permeabilization and apoptosis.
In particular, we have evaluated: 1) histological damage 2) pancreatic digestive enzymes levels 3) neutrophil infiltration by MPO activity assay 4) adhesion molecules expression 5) lipid peroxidation by MDA activity assay 6) IκBα degradation and NFκB nuclear translocation 7) cytokine production 8) iNOS expression and nitrosative stress 9) apoptosis.
Mice were sacrificed 6h or 24h after caerulein-administration. CR3294 was given 1h prior to caerulein-injection and, for animals sacrificed at 24h, CR3294 administration was repeated 8h after caerulein-injection. CR3294 was evaluated at 3 dose levels (20, 10 and 2 mg/kg, os).
The inhibitory effects on iNOS expression of CR3294 show that this molecule decreases, in a dose-dependent manner, the general inflammatory process caused by caerulein-injection and avoids the activation of apoptotic pathways.
In conclusion, our results clearly showed that CR3294 plays an important role in the blocking of the acute pancreatitis development.
CR3294, a novel benzamidine derivative, is an inhibitor of pro-inflammatory gene expression with potent ROS-generation preventing effect. In vitro CR3294 prevents ROS formation and is a potent inhibitor of cytokine production. In vivo, CR3294 has been shown to reduce several types of inflammation (general and intestinal inflammation in animal models [1]), which are related to neutrophil cell influx; it reduces also some septic shock markers, such as iNOS overproduction or TNF release.
The purpose of this study is to analyze the anti-oxidant effect of CR3294 in an experimental animal model of acute pancreatitis.
Pancreatitis induced by cerulein-administration (50 μg/kg, ip) causes histological damage, accompanied by high serum levels of digestive enzymes, high cytokine production, increased expression of adhesion molecules, neutrophil infiltration, release of pro-inflammatory mediators, as well as mitochondrial permeabilization and apoptosis.
In particular, we have evaluated: 1) histological damage 2) pancreatic digestive enzymes levels 3) neutrophil infiltration by MPO activity assay 4) adhesion molecules expression 5) lipid peroxidation by MDA activity assay 6) IκBα degradation and NFκB nuclear translocation 7) cytokine production 8) iNOS expression and nitrosative stress 9) apoptosis.
Mice were sacrificed 6h or 24h after caerulein-administration. CR3294 was given 1h prior to caerulein-injection and, for animals sacrificed at 24h, CR3294 administration was repeated 8h after caerulein-injection. CR3294 was evaluated at 3 dose levels (20, 10 and 2 mg/kg, os).
The inhibitory effects on iNOS expression of CR3294 show that this molecule decreases, in a dose-dependent manner, the general inflammatory process caused by caerulein-injection and avoids the activation of apoptotic pathways.
In conclusion, our results clearly showed that CR3294 plays an important role in the blocking of the acute pancreatitis development.
1) Letari O. et al (2009) – Cancer Chemother Pharmacol, 66(5):819-82