| Backgrounds:Severe acute pancreatitis (SAP) is a complex pathogenesis,disease risks complications,acute abdomen,it’s mortality can be as high as 10 to 30%.The mai: reason for SAP is its pathogenesis is not entirely clear.The SAP’s current researc focuses on self-trypsin digestion,excessive activation of white blooi cells,microcirculation, hyperlipidemia, bacterial infection of these points.With depth study of the extent and level of research to improve the " calciur overload "theory increasingly attracted people’s attention.The important role of Ca2+ i the life activity of cells are well known. It regulates many important physiologica activities.Tumor necrosis factor -α (TNF-α) is a cytokine produced by a variety c inflammatory cells.Numerous studies show that its expression level can be used as a important indicator of pancreatic tissue injury.Animal experiments and clinical observations show that:The expression α TNF-α and NF-κB rapid increase in the incidence of SAP in the short time.Th incidence of their expression is proportional to the amount of pancreatitis. When a lc of accumulation of Ca2+ in the cell, it will cause changes in cell function and eve cause cell death,the current "calcium overload" is considered to be a commo pathway of cell damage.Our experimental observations the expression of NF-κB an TNF-α,and degree of pancreas pancreatic intracellular calcium overload in th pathogenesis within SAP."exploratory study Calcium overload" and "inflammator factor" whether the expression is relevant, that is, whether the pancreatic cell increased Ca2+ concentration can lead to increased expression of inflammator cytokines."San Qi" as a characteristic plant in Yunnan Province,is one of the most preciou traditional Chinese medicine.The Panax notoginseng saponins (panax notoginsen saponins, PNS),the extract of San Qi, are also widely used in a variety of clinica disorders.For pancreatitis,PNS applied more frequently,but the current mechanism for the treatment of pancreatitis is still stuck in terms of " inflammatory factors," oxygen free radicals "etc,for the " calcium overload" has not been involved.1,4, trisphosphate (inositol 1,4,5-trisphosphate, IP3) are intracellular second messengers intracellular free Ca2+regulation plays an important role,the impact of PNS treatmer mechanism for SAP currently in the " calcium overload " has not yet been reported.The purpose:This paper study the relevance of extent of pancreatic cells SAP"calciun overload"and the inflammatory cytokines TNF-α and NF-κB expression;PNS furthe study the protective effect of severe acute pancreatitis as well as on the impact α acinar cell calcium overload.Methods:Healthy adult male Sprague-Dawley (SD) rats..weighing 200～250g, wer randomly divided into three groups.First sham operation group (SO group) Gently flip the duodenum and pancreas after laparotomy, the abdomen was closed; second SAP group:5%sodium taurocholate(1.0mL/kg) injected retrograd< cholangiopancreato-graphy,inducing SAP model. ③PNS group (panax notogi nsenosides pre-intervention group):Before pancreatitis modeling, intraperitoni al injection of PNS (50 mg/ml Xuesetong Injection (0.1ml/100g)).1h,establishe model of acute pancreatitis after injection method in accordance with the SA1 group.Rats in each group respectively after 4h,8h,12h,24h sacrificed to col ect serum,pancreas specimens,calculate the amount of ascites.Biochemical meth ds were measured serum amylase (AMS) activity and serum Ca2+ concentration ELISA assay pancreas IP3 levels.HE staining of the pancreas,light microscopy f pancreatic pathology score.Using flow cytometry (Flow Cytometry/financa capacity model, FCM) detection of intracellular Ca2+ fluorescence intensity (fli orescence intensity, FI), on behalf of the fluorescence intensity using a cell coi centration of Ca2+.Immunohistochemical SP method pancreatic tissue TNF-α am expression of NF-κB. Spearman analysis the correlation of intracellular Ca2+ concentration and TNF-α, NF-κB expression of pancreatic tissue.Results:1. Serum amylase (AMS):Three groups of rats AMS is a significant differenc between the groups at each time point.The AMS of SAP group and PNS group wer significantly higher (P<0.01), the SAP group at each time point comparison was nc statistically significant (P> 0.05) between the PNS group.2. Serum Ca2+:SAP group and PNS group on the 4h,8h,12h time point serum Ca2 concentration is lower than the SO group,the difference was statistically significant ( <0.01),while the SAP group were compared with PNS,PNS group is higher in SA’ group,but the difference was not significant (P> 0.05).When 24h, SAP group and PN! group higher in SO group,but the difference was not statistically significant amon the three groups there was no significant difference (P> 0.05).3. Pancreatic tissue IP3:SAP group,PNS group two groups at each time point IF content than the same time the SO group was significantly higher (P<0.05).SAP grou and PNS group pairwise comparison,4h,8h time point there was no significar difference (P>0.05),IP3 content 12h and 24h, with significant difference,PNS grou were lower than the SAP group (P<0.05)4. Ascites:SAP group and PNS group modeled after open abdominal cavity at a tim point were seen a lot of bloody ascites.Of which the first two groups were compare with SO significant difference (P<0.05).At 4h, between SAP and PNS group was nc significantly different.was not statistically significant (P>0.05).These two groups i 8h,12h,24h comparisons have significant differences(P<0.05), significantly reducin the PNS group than in SAP group ascites.5. Intracellular Ca2+ fluorescence intensity:Using a fluorescence microscop observation shows that pancreatic acinar cells of the developing calcium, showin green. By intracellular Ca2+ fluorescence intensity (Fluorescence Intensity, FI) 1 reflect the intracellular Ca2+ concentration size. Statistics drawn:at this point in tim 4h, SAP group and PNS group were higher than SO group, statistically significant but no statistically significant difference (P>0.05) between SAP group and PNS grou] Between 8h,12h,24h three time points, three groups of rats cell Ca2+ concentratioi were statistically significant (P<0.05). SAP group than in PNS group intracellula Ca2+ fluorescence intensity was significantly higher (P<0.05), with a statistically significant difference.6. Pancreatic histopathologic lesions:SAP group and PNS group pathology scre was significantly higher than the SO group (P<0.05), no statistically significan difference (P between 4h SAP group and PNS group> 0.05). In 8h,12h,24h three time points pathology score analysis shows:PNS group were significantly lower (I <0.05) than the SAP group7. The expression of TNF-α:the pancreatic tissue by immunohistochemistry afte: treatment, showing brown in the cytoplasm, we examined the average optical density by Image pro plus software, reflecting the expression of TNF-a pancreatic tissue by an average apparent density values.Data Tip:are statistically among the three group: for each time point TNF-α average apparent density difference (P<0.05), the average apparent density of SAP and PNS group than SO group were significantly increased In 4h,8h,12h three time points, were significantly lower than the Group SAP anc Group PNS were statistically difference between the two.8. Correlation:Application Spearman rank correlation analysis and Pearsor correlation analysis showed that:in the rat pancreas cells at different time points Ca2-concentrations were positively correlated (P<0.05) and the expression of TNF-α ra pancreatic tissue, intracellular Ca2+ concentration and increased expression of TNF-c correlated.Conclusion:SAP, the pancreatic cells occur "calcium overload", the expression of TNF-α "calcium overload" the degree of correlation between the presence of inflammatory mediators; Panax notoginseng saponins can inhibit the expression of IP3, redu pancreatic cells, "calcium overload" pancreatic tissue protective effects. |
PDF Full Text Request