Effect of BN52021 on NFκ-Bp65 expression in pancreatic tissues of rats with severe acute pancreatitis

AIM： To investigate dynamic changes and significance of expression of NF-κBp65 in pancreatic tissues of rats with severe acute pancreatitis （SAP）, as well as BN52021 effects.
METHODS： Wistar male rats were randomly divided into negative control group （NC group, n = 60）, SAP-model group （SAP group, n = 60）, and BN52021-treated group （BN group, n = 60）, and each of the above groups was respectively divided into 6 subgroups at different time points after operation （1 h, 2 h, 3 h, 6 h, 12 h, and 24 h） （n = 10）. By RT-PCR and Western blot, NF-κBp65 mRNA and its protein expression in pancreatic tissues of rats were detected respectively.
RESULTS： The expression of NF-κBp65 mRNA dynamically changed in both SAP groups and BN groups. The mRNA level was higher in SAP groups than NC groups at 2 h, 3 h, 12 h, and 24 h after operation （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and higher in BN groups than SAP group at 1 h （P 〈 0.05）. The NF-κBp65 protein level was higher in SAP groups than NC groups at 1 h, 3 h, and 6 h （P 〈 0.01）, and 2 h, 12 h, and 24 h （P 〈 0.05）, higher in BN groups than NC groups at all time points （P 〈 0.05）, and lower in BN groups than SAP groups at 1 h, 3 h, and 6 h （P 〈 0.05）.
CONCLUSION： The expression of NF-κBp65 in pancreatic tissues is dynamically changed and the changes play an important role in pathogenesis of SAR BN52021 exerts therapeutic effects through reducing the expression level of NF-κBp65 protein in the early stage of SAR     

Experimental study of therapeutic efficacy of Baicalin in rats with severe acute pancreatitis

AIM： To observe the therapeutic efficacy of Baicalin in rats with severe acute pancreatitis （SAP） and explore its therapeutic mechanisms. METHODS： The SAP rat models were randomly divided into the model control group, Baicalin treatment group, octreotide treatment group and sham operation group. All groups were randomly subdivided into 3 h, 6 h and 12 h groups with 15 rats in each group. The survival, ascites volume and pathological changes of pancreas in all rats were observed at different time points after operation. The plasma amylase content and serum TNF-α, IL-6, malonaldehyde （MDA） and PLA2 contents were also determined. RESULTS： The survival was not obviously different between the treated groups, and was significantly higher in treated groups at 12 h compared to the model control group （P 〈 0.05, 15 vs 10）. The ascites/body weight ratio at 3 h and 6 h was significantly lower in Baicalin treatment group compared to the model control group and octreotide treatment group （P 〈 0.05, 1.00 vs 2.02 and 1.43 and P 〈 0.001, 2.29 （1.21） vs 2.70 （0.80） and 2.08 （2.21）, respectively）. The contents of amylase, TNF-α, IL-6, MDA and PLA2 were significantly lower in the treated groups than in the model control group （P 〈 0.05, 4342 vs 5303, 5058 vs 6272 in amylase, P 〈 0.01, 21.90 vs 36.30, 23.80 vs 39.70, 36 vs 54.35 in MDA and 56.25 vs 76.10 in PIA2, or P 〈 0.001, 65.10 and 47.60 vs 92.15 in TNF-α, 3.03 vs 5.44, 2.88 vs 6.82, 2.83 vs 5.36 in IL-6, respectively）. The pathological scores of pancreas in the treated groups were significantly lower than that in the model control group （P 〈 0.05, 9.00 vs 10.05, 6.00 vs 9.00, 8.00 vs 10.05）, but no marked difference was found between the treated groups. CONCLUSION： The Baicalin injection has significant therapeutic effects on SAP rats, its effects are similar to those of octreotide. The Baicalin injection is also cheap and has a big application range, quite hopefully to be used in clinical treatment of SAP.     

Effects of large dose of dexamethasone on inflammatory mediators and pancreatic cell apoptosis of rats with severe acute pancreatitis

AIM： To investigate the influence of high dose of dexamethasone on inflammatory mediators and apoptosis of rats with severe acute pancreatitis （SAP）.
METHODS： SAP rats were randomly assigned to the model group and treatment group while the normal rats were assigned to the sham operation group. The mortality, ascite volumes, ascites/body weight ratio and pancreas pathological changes of all rats were observed at 3, 6 and 12 h after operation. Their contents of amylase and endotoxin in plasma and contents of tumor necrosis factor （TNF-α）, phospholipase A2 （PLA2） and IL-6 in serum were also determined. The microarray sections of their pancreatic tissues were prepared, terminal transferase dUTP nick end labeling （TUNEL） staining was performed and apoptotic indexes were calculated.
RESULTS： There was no marked difference between treatment group and model group in survival. The contents of amylase and endotoxin in plasma and contents of TNF-α, PLA2 and IL-6 in serum, ascite volumes, ascites/body weight ratio and pancreas pathological scores were all lower in treatment group than in model group to different extents at different time points [P 〈 0.05, 58.3 （26.4） ng/L vs 77.535 （42.157） ng/L in TNF-α content, 8.00 （2.00） points vs 9.00 （2.00） points in pathological score of pancreas respectively; P 〈 0.01, 0.042 （0.018） EU/mL vs 0.056 （0.0195） EU/mL in endotoxin content, 7791 （1863） U/L vs 9195 （1298） U/L in plasma amylase content, 1.53 （0.79） vs 2.38 （1.10） in ascites/body weight ratio, 8.00 （1.00） points vs 11.00 （1.50） points in pathological score of pancreas; P 〈 0.001, 3.36 （1.56） ng/L vs 5.65 （1.08） ng/L in IL-6 content, 4.50 （2.00） vs 7.20 （2.00）, 4.20 （1.60） vs 6.40 （2.30）, 3.40 （2.70） vs 7.90 （1.70） in ascite volumes, respectively]. The apoptotic indexes of pancreas head and pancreas tail were all higher in treatment group than in model group at 6 h [P 〈 0.01, 0.00 （2.00）% vs 0.00 （0.00）%,     

Mechanism of TLR-4/NF-κB pathway in myocardial ischemia reperfusion injury of mouse

Objective: To detect the expression of Toll-like receptor 4(TLR-4) and NF-κB and to discuss the mechanism of TLR-4/NF-κB pathway in the myocardial ischemia reperfusion injury of mouse. Methods: TLR-4 mutant mice and wild homozygous mice were divided into the model group and sham group. Mice in the model group were given the ligation of left anterior descending coronary artery for the modeling, while mice in the sham group were not given the ligation after threading. The cardiac muscle tissues were collected for the morphological observation. The immuno histochemistry was employed to detect the expression of NF-κB, Western blot was used to detect the expression of TLR-4 and ELISA to detect the expression of serum inflammatory factors. Results: The expression of NF-κB in TLR-4 null mice after the myocardial ischemia reperfusion was significantly lower than that in wild homozygous mice. For the model group and sham group, the expression of TLR-4 in wild homozygous mice was all significantly higher than that in TLR-4 null mice, while the expression of TLR-4 in TLR-4 null mice in the model group was significantly higher than that in sham group, with the statistical difference(P0.05). The expression of inflammatory factors in TLR-4 null mice and wild homozygous mice in the model group was significantly higher than that in sham group. The expression of all factors in group A with TLR-4 null was significantly lower than that in group B with wild homozygous type, with the statistical difference(P0.05). Conclusions: TLR-4/NF-κB pathway is closely related to the myocardial ischemia reperfusion injury, which plays its role through the release of inflammatory cytokines.     

Influence of dexamethasone on mesenteric lymph node of rats with severe acute pancreatitis

AIM： To study the influence and mechanisms of dexamethasone on mesenteric lymph node of rats with severe acute pancreatitis （SAP）.
METHODS： The SAP rats were assigned to model, treated or sham-operated groups. The mortality, pathological changes of mesenteric lymph nodes, expression levels of NF-kB, P-selectin, Bax, Bcl-2 and caspase-3 protein and changes in apoptotic indexes in lymph nodes were observed at 3, 6 and 12 h after operation. The blood levels of endotoxin, superoxide dismutase （SOD）, malondialdehyde （MDA）, and endothelin-1 （ET-1） in blood were determined.
RESULTS： SOD content, expression of Bax protein and apoptotic index were significantly higher in the treated group than in the model group at different time points （P 〈 0.05 or P 〈 0.01）. Other blood-detecting indexes and histopathological scores of mesenteric lymph nodes were lower in the treated than in the model group （P 〈 0.05, P 〈 0.01 or P 〈 0.01）. NF-kB protein expression was negative in all groups. Comparing P-selectin and caspase-3 expression levels among all three groups, there was no marked difference between the model and treated group.
CONCLUSION： Dexamethasone can protect mesen-teric lymph nodes. The mechanism may be by reducing the content of inflammatory mediators in the blood and inducing lymphocyte apoptosis.     

Up-regulation of intestinal nuclear factor kappa B and intercellular adhesion molecule-1 following traumatic brain injury in rats

AIM: Nuclear factor kappa B (NF-κB) regulates a large number of genes involved in the inflammatory response to critical illnesses, but it is not known if and how NF-KB is activated and intercellular adhesion molecule-1 (ICAM-1) expressed in the gut following traumatic brain injury (TBI). The aim of current study was to investigate the temporal pattern of intestinal NF-κB activation and ICAM-1 expression following TBI. METHODS: Male Wistar rats were randomly divided into six groups (6 rats in each group) including controls with sham operation and TBI groups at hours 3, 12, 24, and 72, and on d 7. Parietal brain contusion was adopted using weight-dropping method. All rats were decapitated at corresponding time point and mid-jejunum samples were taken. NF-KB binding activity in jejunal tissue was measured using EMSA. Immunohistochemistry was used for detection of ICAM-1 expression in jejunal samples. RESULTS: There was a very low NF-κB binding activity and little ICAM-1 expression in the gut of control rats after sham surgery. NF-KB binding activity in jejunum significantly increased by 160% at 3 h following TBI (P<0.05 vs control), peaked at 72 h (500% increase) and remained elevated on d 7 post-injury by 390% increase. Compared to controls, ICAM-1 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 24 h following TBI and maximally expressed at 72 h post-injury (P<0.001). The endothelial ICAM-1 immunoreactivity in jejunal mucosa still remained strong on d 7 post-injury. The peak of NF-κB activation and endothelial ICAM-1 expression coincided in time with the period during which secondary mucosal injury of the gut was also at their culmination following TBI. CONCLUSION: TBI could induce an immediate and persistent up-regulation of NF-κB activity and subsequent up-regulation of ICAM-1 expression in the intestine. Inflammatory response mediated by increased NF-κB activation and ICAM-1 expression may play an important role in the pathogenesis of acute gut mucosal injury following TBI.     

Protective effect of RadixAstragali injection on immune organs of rats with obstructive jaundice and its mechanism

AIM： To observe the protective effect of RadixAstragali injection on immune organs （lymph nodes, spleen and thymus） of rats with obstructive jaundice （OJ） and its mechanism.
METHODS： SD rats were randomly divided into sham-operation group, model control group and Radix Astragali treatment group. On days 7, 14, 21 and 28 after operation, mortality rate of rats, pathological changes in immune organs, expression levels of Bax and nuclear factor （NF）-κB p65 proteins, apoptosis indexes and serum tumor necrosis factor （TNF）-α level in spleen and thymus were observed, respectively.
RESULTS： Compared to model control group, the number of dead OJ rats in Radix Astragali treatment group decreased （P 〉 0.05）. The TNF-α level （27.62 ± 12.61 vs 29.55± 18.02, 24.61 ± 9.09 vs 31.52± 10.95） on days 7 and 21, the pathological severity score for spleen [0.0 （0.0） vs 0.0 （2.0） on days 7 and 14 and for lymph nodes [0.0 （1.0） vs 1.0 （2.0）, 1.0 （0.0） vs 2.0 （1.0）] on days 21 and 28, the product staining intensity and positive rate of Bax protein in spleen [0.0 （0.0） vs 1.0 （2.0）, 0.0 （1.0） vs 2.0 （1.5） and thymus [0.0 （0.0） vs 1.0 （2.0）, 0.0 （1.0） vs 2.0 （1.5）] on days 14 and 28, the apoptotic indexes [0.0 （0.0） vs 0.0 （0.01）] in spleen and thymus [0.0 （0.0） vs 0.0 （0.01） on days 14 and 21 were significantly lower in Radix Astragali treatment group than in model control group （P 〈 0.05）.
CONCLUSION： Radix Astragali has protective effects on immune organs of OJ rats by relieving the pathological changes in immune organs, reducing TNF-α level and inhibiting Bax expression and apoptosis in spleen and thymus.     

Effect of Danshen on apoptosis and NF-кB protein expression of the intestinal mucosa of rats with severe acute pancreatitis or obstructive jaundice

BACKGROUND:Intestinal mucosa injury in cases of severe acute pancreatitis(SAP) or obstructive jaundice(OJ) is one of the main reasons for the accelerated aggravation of these diseases.Besides being an organ to digest and absorb nutrients,the intestine is also a unique immune organ.When SAP and OJ develop,the destruction of the intestinal mucosa barrier is an important contributing factor for the development of bacterial translocation,systemic inflammatory response syndrome,and multiple organ dysfunction syndrome.It is important to protect the intestinal mucosa in the therapy for SAP and OJ.In this study,we determined the effect of Radix Salviae Miltiorrhizae(Danshen) injection on apoptosis and NF-κB P65 protein expression in the intestinal mucosa of rats with SAP or OJ,and explored the protective mechanism of Danshen in their mucosa.METHODS:Sprague-Dawley rats were used in the SAP and OJ experiments.These rats were randomly divided into shamoperated,model control,and treated groups.At various times after operation,the mortality rates were calculated.Subsequently,the rats were killed to assess the pathological changes,the expression levels of Bax and NF-κB proteins,and the apoptosis indices in the intestinal mucosa.RESULTS:Compared to the corresponding model control group,the number of SAP or OJ rats that died in the treated group decreased but showed no statistically significant difference.At all time points after operation,there was no significant difference between the treated and model control groups in the staining intensity as well as the product of staining intensity and positive staining rate of Bax protein in the intestinal mucosa of SAP and OJ rats.At 3 hours after operation,the apoptosis index of the intestinal mucosa of SAP rats in the treated group was lower than that in the model control group(P0.01).At 12 hours after operation in SAP rats and 28 days after operation in OJ rats,the staining intensity as well as the product of staining intensity and positive staining rate of NF-κB protein of the intestinal mucosa in the treated group were lower than those in the model control group(P0.01).CONCLUSION:Danshen exerts protective effects on the intestinal mucosa of SAP and OJ rats perhaps by inhibiting apoptosis and down-regulating NF-κB protein.     

Heme oxygenase-1 induction by hemin protects liver cells from ischemia/reperfusion injury in cirrhotic rats

AIM： To investigate the potential protective effect of HO-1 on cirrhotic liver cells in rats.
METHODS： Male Wistar rats included in the current study were randomly divided into 5 groups as follows： normal （N） group; liver cirrhotic （LC） group; sham （S） group; I/R group and I/R ＋ hemin group. The model for inducing liver cirrhosis in rats was established according to a previously published protocol. Following this the segmental hepatic ischemia reperfusion operation was carried out. The rats were treated with 30 l~mol/kg hemin （HO-1 inducer, ferric portoporphyrin IX chloride） i.p. or 0.9% NaCI （control） 24 h and 12 h before hepatic ischemia for 30 min or sham laparotomy. Blood was collected for serum enzymatic measurement 6 and 12 h after reperfusion or sham laparotomy. HO-1, NF-κB and caspase-3 expressions were assessed by immunohistochemical analysis.
RESULTS： The expressions of proteins are inversely correlated to the gray values. HO-1 expression in the I/R ＋ hemin group was increased significantly than I/R group at 6 h and 12 h after hepatic I/R （6 h： 112.0± 8.3 vs 125.1± 5.7, P 〈 0.01; 12 h： 120.8± 11.0 vs 132.4 ± 6.2, P 〈 0.01）. Hemin improved serum manganese superoxide dismutase （MnSOD） （6 h： 131.3 ± 17.6 vs 107.0 ± 13.9, P 〈 0.01; 12 h： 141.4 ：E 12.5 vs 118.3± 10.2, P 〈 0.01）, lessened liver cell injury, decreased caspase-3（6 h： 166.7 ± 8.1 vs 145.5 ± 14.6, P 〈 0.01; 12 h： 172.8± 3.8 vs 148.0 ±6.5, P 〈 0.01） and NF-κB expression （6 h： 150.2 ± 8.6 vs 139.7 ±6.0, P 〈 0.01; 12 h： 151.1 ± 5.9 vs 148.1± 5.3, P 〉 0.05） and serum alanine aminotransferase （ALT） （6 h： 413.3± 104.1 vs 626.8 ±208.2, P 〈 0.01; 12 h： 322.2 ± 98.8 vs 425.8 ± 115.4, P 〈 0.05）, aspartate aminotransferase （AST） （6 h： 665.2 ± 70.1 vs 864.3± 70.4, P 〈 0.01; 12 h： 531.1 ± 98.6 vs 664.4± 115.6, P 〈 0.01）, malondialdehyde （MDA） levels （6 h： 11.1 ± 2.17 vs 13.5 ±2.01, P 〈 0.01; 12 h： 9.36     

Influence of dexamethasone on inflammatory mediators and NF-кB expression in multiple organs of rats with severe acute pancreatitis

AIM: To observe the therapeutic effects of dexamethasone on rats with severe acute pancreatitis (SAP) and investigate the influences of dexamethasone on the inflammatory mediators and NF-κB expression in multiple organs of SAP rats as well as the mechanisms involved.METHODS: Ninety Sprague-Dawley (SD) rats with SAP were randomly divided into the model group (n = 45) and dexamethasone treatment group (n = 45), and another 45 rats were selected for the sham operation group. All groups were randomly subdivided into the 3 h, 6 h and 12 h groups, each group containing 15 rats.The survival of all groups and pathological changes of multiple organs (liver, kidney and lung) were observed at different time points after the operation. The pathological score of multiple organs was carried out, followed by the determination of amylase, endotoxin and TNF-α contents in blood. The tissue microarray was used to detect the expression levels of NF-κB p65 protein in multiple organs.RESULTS: There was no marked difference between the model group and treatment group in the survival rate. The amylase contentof the treatment group was significantly lower compared to the model group at 12 h (P ＜ 0.01, 7791.00 vs 9195.00). Moreover, the endotoxin and TNF-α levels of the treatment group were significantly lower than that of the model group at 6 h and 12 h (P ＜ 0.01, 0.040 vs 0.055, 0.042 vs 0.059 and P ＜ 0.05, 58.30 vs 77.54, 38.70 vs 67.30, respectively).Regarding the changes in liver NF-κB expression, the model group significantly exceeded the sham operation group at 3 h (P ＜ 0.01, 1.00 vs 0.00), and the treatment group significantly exceeded the sham operation group at 12 h (P ＜ 0.01, 1.00 vs 0.00), whereas no marked difference was observed between the model group and treatment group at all time points. The kidney NF-κB expression level in the treatment group significantly exceeded the model group (P ＜ 0.05, 2.00 vs 0.00) and the sham operation group (P ＜ 0.01, 2.00 vs 0.00) at 12 h.No NF-κB expression in the lung was found in any group.CONCLUSION: Dexamethasone can lower the amylase,endotoxin and TNF-α levels as well as mortality of SAP rats. NF-κB plays an important role in multiple organ injury. Further studies should be conducted to determine whether dexamethasone can ameliorate the pathological changes of multiple organs by reducing the NF-κB expression in the liver and kidney. The advantages of tissue microarrays in pancreatitis pathological examination include time- and energy- saving, and are highly efficient and representative. The restriction of tissue microarrays on the representation of tissues to various extents due to small diameter may lead to the deviation of analysis.     

Pioglitazone attenuates the severity of sodium taurocholate-induced severe acute pancreatitis

AIM： To determine the effect of pioglitazone, a specific peroxisome proliferator-activated receptor-γ, （PPARγ） ligand, on development of severe acute pancreatitis （SAP） and expression of nuclear factor-kappa B （NF-κB） and intercellular adhesion molecule-1 （ICAM-1） in the pancreas. METHODS： Male Sprague-Dawley （SD） rats （160-200 g） were randomly allocated into three groups （n = 18 in each group）： severe acute pancreatitis group, pioglitazone group, sham group. SAP was induced by retrograde infusion of 1 mL/kg body weight 5% sodium taurocholate （STC） into the biliopancreatic duct of male SD rats. Pioglitazone was injected intraperitoneally two hours piror to STC infusion. Blood and ascites were obtained for detecting amylase and ascitic capacity. Pancreatic wet/dry weight ratio, expression of NF-κB and ICAM-1 in pancreatic tissues were detected by immunohistochemical staining. Pancreatic tissue samples were stained with hematoxylin and eosin （HE） for routine optic microscopy. RESULTS： Sham group displayed normal pancreatic structure. SAP group showed diffuse hemorrhage, necrosis and severe edema in focal areas of pancreas. There was obvious adipo-saponification in abdominal cavity. Characteristics such as pancreatic hemorrhage, necrosis, severe edema and adipo-saponification were found in pioglitazone group, but the levels of those injuries were lower in pioglitazone group than those in SAP group. The wet/dry pancreatic weight ratio, ascetic capacity, serum and ascitic activities of anylase in the SAP group were significantly higher than those in the sham group and pioglitazone group respectively （6969.50 ± 1368.99 vs 2104.67 ± 377.16, 3.99 ± 1.22 vs 2.48 ± 0.74, P 〈 0.01 or P 〈 0.05）. According to Kusske criteria, the pancreatic histologic score showed that interstitial edema, inflammatory infiltration, parenchyma necrosis and parenchyma hommorrhage in SAP group significantly differed from those in the sham group and pioglitazone group （7.17 ± 1.83 vs     

Application of Tissue Microarrays to Study the Influence of Dexamethasone on NF-κB Expression of Pancreas in Rat with Severe Acute Pancreatitis

To discuss the influence of dexamethasone on NF-κB expression of pancreas in rat with severe acute pancreatitis (SAP). Ninety rat SAP models were divided into the model group and dexamethasone treatment group with 45 rats in each group; another healthy 45 rats were selected to be the sham operation group. The groups were divided into the 3, 6 and 12 h group with 15 rats in each group. The survivals, pancreas pathological changes were observed 3, 6 and 12 h after operation. The changes in expression levels of NF-κB protein of pancreas tissue microarray were observed. The treatment group was significantly lower than the model group at 3 and 6 h (P < 0.05) and than the model group at 12 h in pancreas pathological scores (P < 0.01). The expression level of NF-κB protein of pancreas head of the treatment group was significantly less than that of the model group at 3 h (P < 0.01). The alleviation of pancreatic tissue injury by dexamethasone during SAP might be closely related to its role in inhibiting NF-κB expression and regulating cytokines. The advantages of tissue microarrays in pancreatitis pathological examination include time and energy savings, high efficiency and representative results.     

Preparation method of an ideal model of multiple organ injury of rat with severe acute pancreatitis

AIM To establish an ideal model of multiple organ injury of rats with severe acute pancreatitis (SAP).METHODS SAP models were induced by retrograde injection of 0.1 mL/100 g 3.5% sodium taurocholate into the biliopancreatic duct of Sprague-Dawley rats.The plasma and samples of multiple organ tissues of rats were collected at 3, 6 and 12 h after modeling. The ascites volume, ascites/body weight ratio, and contents of amylase, endotoxin, endothelin-1 (ET-1), nitrogen monoxidum (NO), phospholipase A2 (PLA2), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) in plasma were determined. The histological changes of multiple organs were observed under light microscope.RESULTS The ascites volume, ascites/body weight ratio, and contents of various inflammatory mediators in blood were higher in the model group than in the sham operation group at all time points [2.38 (1.10), 2.58(0.70), 2.54 (0.71) vs 0.20 (0.04), 0.30 (0.30), 0.22 (0.10)at 3, 6 and 12 h in ascites/body weight ratio; 1582 (284),1769 (362), 1618 (302) (U/L) vs 5303 (1373), 6276(1029), 7538 (2934) (U/L) at 3, 6 and 12 h in Amylase;0.016 (0.005), 0.016 (0.010), 0.014 (0.015) (EU/mL) vs0.053 (0.029), 0.059 (0.037), 0.060 (0.022) (EU/mL)at 3, 6 and 12 h in Endotoxin; 3.900 (3.200), 4.000(1.700), 5.300 (3.000) (ng/L) vs 41.438 (37.721), 92.151(23.119), 65.016 (26.806) (ng/L) at 3, 6 and 12 hin TNF-α, all P ＜ 0.01]. Visible congestion, edema and lamellar necrosis and massive leukocytic infiltration were found in the pancreas of rats of model group. There were also pathological changes of lung, liver, kidney,ileum, lymphonode, thymus, myocardium and brain.CONCLUSION This rat model features reliability,convenience and a high achievement ratio. Complicated with multiple organ injury, it is an ideal animal model of SAP.     

Activated protein C,an anticoagulant polypeptide,ameliorates severe acute pancreatitis via regulation of mitogen-activated protein kinases

Background Our aim was to investigate the changes of mitogen-activated protein kinases (MAPKs) by activated protein C (APC) treatment in rats with severe acute pancreatitis (SAP), and relate them to changes in SAP severity, thus providing evidence for developing clinical therapies. Methods Sprague-Dawley rats were given an intravenous injection of saline (SAP group), APC (50 μg/kg or 10 μg/kg), or CNI1493 just before SAP induction. One group of rats underwent a sham operation (control group). Experimental samples were harvested 16 h after SAP induction. The gene expression of pancreatic MAPKs was evaluated by cDNA microarrays. The mRNA and protein/phosphorylated protein levels of p38 MAPK, extracellular signal-regulated protein kinase (ERK) 1/2, and c-Jun N-terminal kinase (JNK) and the protein levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined in pancreatic tissue. The severity of disease was evaluated by pancreatic histology, the pancreatic wet/dry weight ratio, and the serum amylase level. Results In rats treated with APC (50 μg/kg) or CNI1493, the severity of pancreatitis and expression of pancreatic TNF-α and IL-1β proteins were attenuated by the decreased expression and activity of p38 MAPK and JNK (vs. the SAP group, P < 0.01). The expression and activity of ERK1/2 were increased in APC-treated rats, especially in the group treated with APC 50 μg/kg (vs. the SAP or CNI1493-treated group, P < 0.01, respectively). Conclusions Inhibition of expression of pancreatic p38 MAPK and JNK and upregulation of ERK1/2 expression by APC treatment may protect against pancreatic injury, thus ameliorating severity of the disease.     

Effect of baicalin and octreotide on the expression levels of P-selectin protein in multiple organs of rats with severe acute pancreatitis

Aim: To investigate the effect of baicalin and octreotide on the expression levels of P‐selectin protein in multiple organs of rats with severe acute pancreatitis and explore the underlying mechanism. Methods: Rats were randomly divided into sham‐operated, model control, baicalin‐treated and octreotide‐treated groups. At 3, 6 and 12 h after operation, the mortality rates of rats, the contents of plasma endotoxin as well as serum NO and ET‐1, the pathological changes in multiple organs, and the expression levels of P‐selectin protein in each group were observed. Results: At 12 h after operation, the mortality rates of rats in treated groups were significantly lower than that in the model control group (P < 0.05), and the pathological severity scores in multiple organs in treated groups were also significantly lower than those in the model control group (P < 0.05). The contents of plasma endotoxin, serum PLA₂ (at 6 and 12 h after operation), ET‐1 and NO (at 3 and 12 h after operation) in treated groups were significantly lower than those in the model control group (P < 0.05, P < 0.01 or P < 0.001). In the baicalin‐treated group, the expression levels of P‐selectin protein in liver (at 3 h after operation), kidney (at 3 and 6 h after operation), pancreas, lung and spleen were significantly lower than those in the model control group (P < 0.01). In the octreotide‐treated group, the expression levels of this protein in lung, intestinal mucosa (at 6 and 12 h after operation), lymph nodes (at 3 and 6 h after operation), spleen and thymus were significantly lower than those in the model control group (P < 0.05). Additionally, the products of the staining intensity and positive rate of P‐selectin protein in pancreas, spleen (at 3 h after operation), intestinal mucosa (at 6 h after operation), thymus (at 6 h after operation) and lung (at 6 h after operation) in treated groups were significantly lower than those in the model control group (P < 0.05). Conclusion: Both baicalin and octreotide can exert some protective effects on multiple organs and the former is superior to the latter in protecting pancreas. Furthermore, decreasing the expression levels of P‐selectin protein in these organs is one of the possible mechanisms.     

Protective effects and mechanisms of Baicalin and octreotide on renal injury of rats with severe acute pancreatitis

AIM: To investigate the protective effects and mechanisms of Baicalin and octreotide on renal injury of rats with severe acute pancreatitis (SAP). METHODS: One hundred and eighty SD rats were randomly assigned to the model group, Baicalin-treated group, octreotide-treated group and sham operation group. The mortality, plasma endotoxin level, contents of blood urea nitrogen (BUN), creatinine (CREA), phospholipase A2 (PLA2), nitrogen monoxide (NO), tumor necrosis factor (TNF)-alpha, IL-6 and endothelin-1 (ET-1) in serum, expression levels of renal Bax and Bcl-2 protein, apoptotic indexes and pathological changes of kidney were observed at 3, 6 and 12 h after operation. RESULTS: The renal pathological changes were milder in treated group than in model group. The survival at 12 h and renal apoptotic indexes at 6 h were significantly (P<0.05) higher in treated group than in model group [66.67% vs 100%; 0.00 (0.02)% and 0.00 (0.04)% vs 0.00 (0.00)%, respectively]. The serum CREA content was markedly lower in octreotide-treated group than in model group at 3 h and 6 h (P<0.01, 29.200+/-5.710 micromol/L vs 38.400+/-11.344 micromol/L; P<0.05, 33.533+/-10.106 micromol/L vs 45.154+/-17.435 micromol/L, respectively). The expression level of renal Bax protein was not significantly different between model group and treated groups at all time points. The expression level of renal Bcl-2 protein was lower in Baicalin-treated group than in model group at 6 h [P<0.001, 0.00 (0.00) grade score vs 3.00 (3.00) grade score]. The Bcl-2 expression level was lower in octreotide-treated group than in model group at 6 h and 12 h [P<0.05, 0.00 (0.00) grade score vs 3.00 (3.00) grade score; 0.00 (0.00) grade score vs 0.00 (1.25) grade score, respectively]. The serum NO contents were lower in treated groups than in model group at 3 h and 12 h [P<0.05, 57.50 (22.50) and 52.50 (15.00) micromol/L vs 65.00 (7.50) micromol/L; P<0.01, 57.50 (27.50) and 45.00 (12.50) micromol/L vs 74.10 (26.15) micromol/L, respectively]. The plasma endotoxin content and serum BUN content (at 6 h and 12 h) were lower in treated groups than in model group. The contents of IL-6, ET-1, TNF-alpha (at 6 h) and PLA2 (at 6 h and 12 h) were lower in treated groups than in model group [P<0.001, 3.031 (0.870) and 2.646 (1.373) pg/mL vs 5.437 (1.025) pg/mL; 2.882 (1.392) and 3.076 (1.205) pg/mL vs 6.817 (0.810) pg/mL; 2.832 (0.597) and 2.462 (1.353) pg/mL vs 5.356 (0.747) pg/mL; 16.226 (3.174) and 14.855 (5.747) pg/mL vs 25.625 (7.973) pg/mL; 18.625 (5.780) and 15.185 (1.761) pg/mL vs 24.725 (3.759) pg/mL; 65.10 (27.51) and 47.60 (16.50) pg/mL vs 92.15 (23.12) pg/mL; 67.91+/-20.61 and 66.86+/-22.10 U/mL, 63.13+/-26.31 and 53.63+/-12.28 U/mL vs 101.46+/-14.67 and 105.33+/-18.10 U/mL, respectively]. CONCLUSION: Both Baicalin and octreotide can protect the kidney of rats with severe acute pancreatitis. The therapeutic mechanisms of Baicalin and octreotide might be related to their inhibition of inflammatory mediators and induction of apoptosis. Baicalin might be a promising therapeutic tool for severe acute pancreatitis.     

Effect of resveratrol on activation of nuclear factor kappa-B and inflammatory factors in rat model of acute pancreatitis

AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control), severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by irnmunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimrnunoassay. The pathological changes of pancreas and lungs were examined microscopically. RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage, exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group, the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs 2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36%,P<0.01), TNF-α, IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P<0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs 36.23±2.30%), TNF-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs 0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23μg/L). CONCLUSION: The activation of NF-KB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation, alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.