Role Of NO And ICAM-1in Microcirculation Disturbance Of Rats With Severe Acute Pancreatitis

Objectives To establish severe acute pancreatitis animal model and dynamicallyobserve the changes of microcirculation in the early onset of severe acute pancreatitis(SAP). To explore the changes of severe acute pancreatitis microcirculation function indifferent groups and their influence on the body. To investigate the role of nitric oxide(NO) and intercellular adhesion molecules-1(ICAM-1) in the microcirculationdevelopment process of severe acute pancreatitis. This studying can provide a sciencebasis by improving disturbance of microcirculation and the cure rate of SAP.Methods60specific pathogen free (SPF) wistar male rats were divided into fivegroups randomly: blank group (n=12), sham-operated group (n=12), six hours afterpancreatitis group (n=12), twelve hours after pancreatitis group (n=12), twenty-fourhours after pancreatitis group (n=12). Pancreatitis was induced by injection of5%Na-Tc under the membrane of pancreas. The rats were killed at sixth hour afteroperated in sham-operated group. The rats were killed at sixth, twelfth andtwenty-fourth hour after Na-Tc injection in six hours after pancreatitis group, twelvehours after pancreatitis group and twenty-four hours after pancreatitis grouprespectively. The concentration of amylase in serum was measured by holo-automaticbiochemical analysator. Pancreatic tissues were observed by hematoxylin-eosin (HE)staining and were counted for the pancreatic histopathologic scores. We detected thechanges of the hemorheology by using blood viscosity degree instrument. We detectedthe concentration of soluble intercellular adhesion molecules-1(sICAM-1) in serum with ELISA method. We detected the expression of ICAM-1in pancreatic tissue byimmuno-histochemical method. The concentration of NO was measured by nitratereductase method. We detected the serum prothrombin time (PT), the activation partialprothrombin time (APTT), and the concentration of fibrinogen (FIB) and D-Dimer(D-D) with automatic blood coagulation analyzer. All the data were analyzed bystatistics software SPSS17.0. Normal distribution measurement data were shown asmeans±standard deviation (x±s). One-way ANOVA method was applied tocomparison among groups. And we used S-N-K method for binary comparison.Pearson correlation analysis was applied to measure the correlationship. The inspectionstandards was alpha=0.05.Results We successfully established severe acute pancreatitis animal model. The ratsof pancreatitis group exhibited high amylase and pancreatic pathological injury aftermodeling. The pancreatic histopathologic scores in sham-operated group comparedwith blank group was no significant difference (P>0.05). The pancreatichistopathologic scores in six hours after pancreatitis group, twelve hours afterpancreatitis group and twenty-four hours after pancreatitis group compared with blankgroup and sham-operated group were significant difference (P<0.05), and six hoursafter pancreatitis group, twelve hours after pancreatitis group and twenty-four hoursafter pancreatitis group compared with each other were significant difference (P<0.05).The indexes of coagulation system in sham-operated group compared with blank groupwere no significant difference (P>0.05). The indexes of coagulation system in sixhours after pancreatitis group, twelve hours after pancreatitis group and twenty-fourhours after pancreatitis group compared with blank group and sham-operated groupwere significant difference (P<0.05), and six hours after pancreatitis group, twelvehours after pancreatitis group and twenty-four hours after pancreatitis group comparedwith each other were significant difference (P<0.05). The indexes of hemorheology insham-operated group compared with blank group were no significant difference(P>0.05). The indexes of hemorheology in six hours after pancreatitis group, twelvehours after pancreatitis group and twenty-four hours after pancreatitis group comparedwith blank group and sham-operated group were significant difference (P<0.05). Theconcentration of NO and sICAM-1in sham-operated group compared with blank groupwere no significant difference (P>0.05). The concentration of NO and sICAM-1in sixhours after pancreatitis group, twelve hours after pancreatitis group and twenty-fourhours after pancreatitis group compared with blank group and sham-operated group were significant difference (P<0.05). The optical density (OD) of ICAM-1insham-operated group compared with blank group were no significant difference(P>0.05). The optical density (OD) of ICAM-1in six hours after pancreatitis group,twelve hours after pancreatitis group and twenty-four hours after pancreatitis groupcompared with blank group and sham-operated group were significant difference(P<0.05). The pancreatic histopathologic scores, concentration of sICAM-1and NOwere highly positive correlation (r>0.7, P<0.01) with whole blood viscosity, PT, APTT,FIB, D-D and FDP in the course of severe acute pancreatitis.Conclusions There were hemorheology and blood coagulation function disturbancesin the early stage of severe acute pancreatitis. And those disturbances were aggravatedas the illness grew worse. There was a positive correlation between the concentrationof NO and sICAM-1in serum, pancreatic histopathologic scores and the disturbancesof hemorheology and blood coagulation function. These changes can be considered asvery significant experiment indexes which can estimate the severity and the prognosisof severe acute pancreatitis. These changes of microcirculation were aggravated as theillness grew worse and were closely related (r>0.7, P<0.01) to the concentration of NOand sICAM-1in serum.