The Effect Of PNS On Rats Acinar Cell Calcium Overload And NF-κB With Severe Acute Pancreatitis

Objective:To investing the change of the calcium overload and NF-κB expression level after panax notoginseng saponins in treatment of rats with severe acute pancreatitis,and to explore the relationship of calcium overload and NF-κB.Methods:Ninety healthy adult Sprague-Dawley (SD) rats,the weighing about 200～ 250g,were randomly divided into three groups:sham operation group (SO group),severe acute pancreatitis group (SAP group),PNS preconditioning group(PNS group),and there were 30 rats in each group.①The sham operation group(SO group):Gently flip the duodenum and pancreas after laparotomy,then closed the abdomen;②severe acute pancreatitis group (SAP group):Open the abdomen,5%sodium taurocholate(1.0mL/kg) injected retrograde cholangiopancreato-graphy,inducing SAP model. ③PNS preconditioning group(PNS group):Before pancreatitis modeling,intraperitoneal injection of PNS (50 mg/ml Xuesetong Injection (0.1ml/100g)).Afrer 1 hour,established model of acute pancreatitis after injection method in accordance with the SAP group.Rats in the three groups respectively after 8h,12h,24h sacrificed to collect ascites,serum and pancreas specimens.And calculate the amount of ascites;Biochemical methods were measured serum amylase (AMS) activity;HE staining of the pancreas,light microscopy of pancreatic pathology score;Immunohistochemical SP method pancreatic tissue expression of NF-κB;Using flow cytometry (Flow Cytometry/Financal capacity model, FCM) detection of intracellular Ca2+ fluorescence intensity (fluorescence intensity, FI), on behalf of the fluorescence intensity using a cell concentration of Ca2+. Spearman analysis the correlation of intracellular Ca2+ concentration and NF-κB expression of pancreatic tissue.Results:1. Serum amylase (AMS):Three groups of rats AMS is a significant difference between the groups at the each time point(P<0.05).Compared with SO group,SAP group and PNS group were significantly higher,compared statistically significant(P<0.05);Comp-ared with SAP group,the serum AMS of the PNS group were significantly reduced at each time point,compared statistically(P<0.05).2.Ascites:There were large amounts of bloody ascites at 8h,12h,24h in SAP group and PNS group.Compared with SO group,SAP group and PNS group were significantly higher,compared statistically significant(P<0.05);Compared with SAP group,the serum AMS of the PNS group were significantly reduced at each time point,compared statistically(P<0.05).3.Pancreatic histopathologic lesions:Compared with SO group,SAP group and PNS group were significantly higher,compared statistically significant(P<0.05);Compared with SAP group,the serum AMS of the PNS group were significantly reduced at each time point,compared statistically(P<0.05).4.Intracellular Ca2+ fluorescence intensity of pancreatic cells:Using a fluorescence microscope observation shows that pancreatic acinar cells of the developing calcium, showing green.By intracellular Ca2+ fluorescence intensity (Fluorescence Intensity, FI) to reflect the intracellular Ca2+ concentration size.In 8h,12h,24h,the fluorescence intensity of Intracellular Ca2+ were satistically significant in the each group(P<0.05);Compared with SO group,SAP group and PNS group were significantly higher,compared statistically significant(P<0.05);Compared with SAP group,the serum AMS of the PNS group were significantly reduced at each time point,compared statistically(P<0.05).5.The changes of pancreas NF-κB factor level:The pancreatic tissue by immunohistochemistry after treatment, showing tan in the cell,through the Image pro plus software,at the 8h,12h24h,Compared with SO group,SAP group and PNS group were significantly higher,compared statistically significant(P<0.05);Compared with SAP group,the serum AMS of the PNS group were significantly reduced at each time point,compared statistically(P<0.05).6.Correlation analysis:The Spearman correlation analysis shows:at different time points,Ca2+ concentrations were positively correlated and the expression of NF-κB rat pancreatic tissue in the rat pancreas cells(P<0.05), intracellular Ca2+ concentration and increased expression of NF-κB correlated.Conclusion:1.The pancreas intracellular calcium overload and NF-κB factor levels were significantly activated during severe acute pancreatitis,and in the high level expression;2.The level change of acinar cell calcium overload and the NF-κB factor were positively correlated,suggesting that intracellular calcium overload may cause NF-κB factor of high expression in the process of acute pancreatitis,and causing a series of inflamm atoryreaction.3.Panax notoginseng saponins can inhibit the intracellular calcium overload,further reduce inflammation,may help in the treatment of acute pancreatitis.