Influence of dexamethasone on inflammatory mediators and NF-κB 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 content,of 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     

Study on protecting effects of Baicalin and Octreotide on hepatic injury in rats with severe acute pancreatitis

AIM： To investigate the protective effects and mechanisms of Baicalin and Octreotide on hepatic injury in rats with severe acute pancreatitis （SAP）. METHODS： The SAP rat models were prepared and randomly assigned to the model control group, Baicalin treated group, and Octreotide treated group while other healthy rats were assigned to the shamoperated group. Rat mortality, levels of ALT, AST, liver and pancreas pathological changes in all groups were observed at 3, 6 and 12 h after operation. Tissue microarray （TMA） sections of hepatic tissue were prepared to observe expression levels of Bax, Bcl-2 protein and Caspase-3, and changes of apoptotic indexes.RESULTS： Rat survival at 12 h, expression levels of Bax, Caspase-3 protein and apoptotic indexes of liver were all significantly higher in treated groups than in model control group. While the liver and pancreas pathological scores, contents of ALT, AST, and expression levels of Bcl-2 protein were all lower in treated groups than in the model control group. CONCLUSION： Both Baicalin and Octreotide can protect rats with SAP by decreasing the contents of ALT, AST and expression levels of Bcl-2 protein and improving the expression levels of Bax protein, Caspase-3 protein, and inducing apoptosis.     

Adenoviral transfer of human interleukin-10 gene in lethal pancreatitis

AIM: To evaluate the therapeutic effect of adenoviral-vector-delivered human interleukin-10 (hIL-10) gene on severe acute pancreatitis (SAP) rats. METHODS: Healthy Sprague-Dawley (SD) rats were intraperitoneally injected with adenoviral IL-10 gene (AdvhIL-10), empty vector (Adv0) or PBS solution. Blood, liver, pancreas and lung were harvested on the second day to examine hIL-10 level by ELISA and serum amylase by enzymatic assay. A SAP model was induced by retrograde injection of sodium taurocholate through pancreatic duct. SAP rats were then administered with AdvhIL-10, Adv0and PBS solution by a single intraperitoneal injection 20 min after SAP induction. In addition to serum amylase assay, levels of hIL-10 and tumor necrosis factor-α (TNF-α) were detected by RT-PCR, ELISA and histological study. The mortality rate was studied and analyzed by Kaplan-Meier and log rank analysis.RESULTS: The levels of hIL-10 in the pancreas, liver and lung of healthy rats increased significantly after AdvhIL-10 injection (1.42 ng/g in liver, 0.91 ng/g in pancreas); while there was no significant change of hIL-10 in the other two control groups. The concentration of hIL-10 was increased significantly in the SAP rats after AdvhIL-10 injection (1.68 ng/g in liver, 1.12 ng/g in pancreas) compared to the other two SAP groups with blank vector or PBS treatment (P<0.05). The serum amylase levels remained normal in the AdvhIL-10 transfected healthy rats. However, the serum amylase level was significantly elevated in the other two control SAP rats. In contrast, serum amylase was down-regulated in the AdvhIL-10 treated SAP groups.The TNF-α expression in the AdvhIL-10 treated SAP rats was significantly lower compared to the other two control SAP groups. The pathohistological changes in the AdvhIL-10 treated group were better than those in the other two control groups. Furthermore, the mortality of the AdvhIL-10 treated group was significantly reduced compared to the other two control groups (P<0.05). CONCLUSION: Adenoviral hIL-10 gene can significantly attenuate the severity of SAP rats, and can be used in the treatment of acute inflammation process.     

Effects of emodin and baicalein on rats with severe acute pancreatitis

AIM: To investigate the therapeutic effects of emodin in combination with baicalein on severe acute pancreatitis (SAP) rats and to explore the mechanism of SAP. METHODS: A total of 112 SAP rats induced by retrograde injection of 5% sodium taurocholate into the biliary-pancreatic duct, randomly assigned to a untreated group and three treated groups emodin group, combined emodin and baicalein group, and sandostatin group. Meanwhile, another 28 other rats were selected as sham operation (SO) group. There were 28 rats in each group, 8 rats were in 3 and 6 h groups respectively, and 12 rats in 12 h group. At each time-points, survival rates,ascites volumes, pathological lesion scores of pancreas tissues,serum amylase, tumor necrosis factor-α and IL-6 levels were determined as the indexes of therapeutic effects. RESULTS: The survival rate at 12 h was significantly higher in three treated groups than in untreated group.The ascites volume at 12 h was remarkably less in combined and sandostatin groups than in emodin group,but there was no difference between combined group and sandostatin group (P>0.05). Serum amylase levels at all time-points were significantly lower in three treated groups than in untreated group. However, they had no difference among treated groups (P>0.05).Serum TNF-α were lower in three treated groups than in untreated group at all time points. Among the three treated groups, at 6 h, the TNF-α levels of combination and sandostatin groups were lower than those of emodin group. These was no difference between combined and sandostantin. Serum IL-6 concentration at 3 h were lower in combined and sandostatin groups than in untreated group, but at 6 and 12 h they were lower in all treated groups than in untreated group and the combined and sandostatin groups and in emodin group, no difference was found between combined and sandostatin groups at all time-points (P>0.05). The pathological scores of pancreas at all time points were significantly lower in three treated groups than in the untreated group, and at 6, 12 h, the scores of combined and sandostatin groups were lower than in emodin group. There was no difference between combined and sandostatin groups (P>0.05). CONCLUSION: Combination of emodin with baicalein has significant therapeutic effects on SAP rats.     

Adenoviral transfer of human intedeukin-10 gene in lethal pancreatitis

AIM:To evaluate the therapeutic effect of adenoviral-vector-delivered human interleukin-10(hIL-10)gene on severeacute pancreatitis(SAP)rats.METHODS:Healthy Sprague-Dawley(SD)rats wereintraperitoneally injected with adenoviral IL-10 gene(AdvhIL-10),empty vector(Adv0)or PBS solution.Blood,liver,pancreas and lung were harvested on the secondday to examine hIL-10 level by ELISA and serum amylaseby enzymatic assay.A SAP model was induced by retrogradeinjection of sodium taurocholate through pancreatic duct.SAP rats were then administered with AdvhIL-10,Adv0and PBS solution by a single intraperitoneal injection 20 minafter SAP induction.In addition to serum amylase assay,levels of hIL-10 and tumor necrosis factor-α(TNF-α)weredetected by RT-PCR,ELISA and histological study.Themortality rate was studied and analyzed by Kaplan-Meierand log rank analysis.RESULTS:The levels of hIL-10 in the pancreas,liver andlung of healthy rats increased significantly after AdvhIL-10injection(1.42 ng/g in liver,0.91 ng/g in pancreas);whilethere was no significant change of hIL-10 in the other twocontrol groups.The concentration of hIL-10 was increasedsignificantly in the SAP rats after AdvhIL-10 injection(1.68 ng/g in liver,1.12 ng/g in pancreas)compared tothe other two SAP groups with blank vector or PBStreatment(P<0.05).The serum amylase levels remainednormal in the AdvhIL-10 transfected healthy rats.However,the serum amylase level was significantly elevated in theother two control SAP rats.In contrast,serum amylasewas down-regulated in the AdvhIL-10 treated SAP groups.The TNF-α expression in the AdvhIL-10 treated SAP ratswas significantly lower compared to the other two controlSAP groups.The pathohistological changes in the AdvhIL-10 treated group were better than those in the other twocontrol groups.Furthermore,the mortality of the AdvhIL-10 treated group was significantly reduced compared tothe other two control groups(P<0.05).CONCLUSION:Adenoviral hIL-10 gene can significantlyattenuate the severity of SAP rats,and can be used in the treatment of acute inflammation process.     

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）%,     

Effects of Chai-Qin-Cheng-Qi Decoction on cefotaxime in rats with acute necrotizing pancreatitis

AIM： To investigate the effect of Chai-Qin-Cheng-Qi Decoction （CQCQD） on cefotaxime （CTX） concentration in pancreas of rats with acute necrotizing pancreatitis （ANP）. METHODS： Sixty healthy male Sprague-Dawley rats were divided randomly into an ANP group （ANP model ＋ CTX, n = 20）, treatment group （ANP model ＋ CTX ＋ CQCQD, n = 20） and control group （normal rats ＋ CTX, n = 20）. ANP models were induced by retrograde intraductal injection of 3.5% sodium taurocholate （1 mL/kg）, and the control group was injected intraductally with normal saline. All rats were injected introperitoneally with 0.42 g/kg CTX （at 12-h intervals for a continuous 72 h） at 6 h after intraductal injection. Meanwhile, the treatment group received CQCQD （20 mL/kg） intragastrically at 8-h intervals, and the ANP and control group were treated intragastrically with normal saline. At 15 min after the last CTX injection, blood and pancreas samples were collected for the determination of CTX concentration using validated high-performance liquid chromatography. Pathological changes and wet-to-dry-weight （W/D） ratio of pancreatic tissue were examined. RESULTS： Serum CTX concentrations in three groups were not significantly different. Pancreatic CTX concentration and penetration ratio were lower in ANP group vs control group （4.4 ± 0.6 μg/mL vs 18.6±1.7μg/mL, P = 0.000; 5% vs 19%, P = 0.000）, but significantly higher in treatment group vs ANP group （6.4 ± 1.7 μg/mL vs 4.4 ± 0.6 μg/mL, P = 0.020; 7% vs 5%, P = 0.048）. The histological scores and W/D ratio were significantly decreased in treatment group vs ANP and control group. CONCLUSION： CQCQD might have a promotive effect on CTX concentration in pancreatic tissues of rats with ANP.     

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.     

Probiotics improve survival of septic rats by suppressing conditioned pathogens in ascites

AIM: To investigate the benefits of probiotics treatment in septic rats. METHODS: The septic rats were induced by cecal ligation and puncture. The animals of control, septic model and probiotics treated groups were treated with vehicle and mixed probiotics, respectively. The mixture of probiotics included Bifidobacterium longum , Lacto-bacillus bulgaricus and Streptococcus thermophilus . We observed the survival of septic rats using different amounts of mixed probiotics. We also detected the bacterial population in ascites and blood of experimental sepsis using cultivation and real-time polymerase chain reaction. The severity of mucosal inflammation in colonic tissues was determined. RESULTS: Probiotics treatment improved survival of the rats significantly and this effect was dose dependent. The survival rate was 30% for vehicle-treated septic model group. However, 1 and 1/4 doses of probiotics treatment increased survival rate significantly comparedwith septic model group (80% and 55% vs 30%, P < 0.05). The total viable counts of bacteria in ascites decreased significantly in probiotics treated group compared with septic model group (5.20 ± 0.57 vs 9.81 ± 0.67, P < 0.05). The total positive rate of hemoculture decreased significantly in probiotics treated group compared with septic model group (33.3% vs 100.0%, P < 0.05). The population of Escherichia coli and Staphy-lococcus aureus in ascites of probiotics treated group were decreased significantly compared with that of septic model group (3.93 ± 0.73vs 8.80 ± 0.83,P < 0.05; 2.80 ± 1.04 vs 5.39 ± 1.21, P < 0.05). With probiotics treatment, there was a decrease in the scores of inflammatory cell infiltration into the intestinal mucosa in septic animals (1.50 ± 0.25vs 2.88 ± 0.14,P < 0.01). CONCLUSION: Escherichia coli and Staphylococcus aureus may be primary pathogens in septic rats. Probiotics improve survival of septic rats by suppressing these conditioned pathogens.     

Delayed ethyl pyruvate therapy attenuates experimental severe acute pancreatitis via reduced serum high mobility group box 1 levels in rats

AIM： To investigate the effect of delayed ethyl pyruvate （EP） delivery on distant organ injury, survival time and serum high mobility group box 1 （HMGB1） levels in rats with experimental severe acute pancreatitis （SAP）.
METHODS： A SAP model was induced by retrograde injection of artificial bile into the pancreatic ducts of rats. Animals were divided randomly into three groups （n = 32 in each group）： sham group, SAP group and delayed EP treatment group. The rats in the delayed EP treatment group received EP （30 mg/kg） at 12 h, 18 h and 30 h after induction of SAP. Animals were sacrificed, and samples were obtained at 24 h and 48 h after induction of SAP. Serum HMGB1, aspartate arninotransferase （AST）, alanine arninotransferase （ALT）, blood urea nitrogen （BUN）, and creatinine （Cr） levels were measured. Lung wet-to-dry-weight （W/D） ratios and histological scores were calculated to evaluate lung injury. Additional experiments were performed between SAP and delayed EP treatment groups to study the influence of EP on survival times of SAP rats.
RESULTS： Delayed EP treatment significantly reduced serum HMGB1 levels, and protected against liver, renal and lung injury with reduced lung W/D ratios （8.22 ±0.42 vs 9.76 ± 0.45, P 〈 0.01）, pulmonary histological scores （7.1 ± 0.7 vs 8.4 ± 1.1, P 〈 0.01）, serum AST （667 ± 103 vs 1 368 ± 271, P 〈 0.01）, ALT （446 ± 91 vs 653 ± 98, P 〈 0.01） and Cr （1.2 ± 0.3 vs 1.8 ± 0.3, P 〈 0.01） levels. SAP rats had a median survival time of 44 h. Delayed EP treatment significantly prolonged median survival time to 72 h （P 〈 0.01）.
CONCLUSION： Delayed EP therapy protects against distant organ injury and prolongs survival time via reduced serum HMGBllevels in rats with experimental SAP. EP may potentially serve as an effective new therapeutic option against the inflammatory response and multiple organ dysfunction syndrome （MODS） in SAP patients.     

Protective Effects of Baicalin and Octreotide on Multiple Organ Injury in Severe Acute Pancreatitis

Purpose To discuss the application value of Baicalin which is a new drug by comparing the protecting effects of Baicalin and Octreotide on multiple organs (pancreas, liver, kidney, and lung) in Severe acute pancreatitis (SAP). Methods The improved Aho method was adopted to prepare SAP rat models via retrograde injection of 3.5% sodium taurocholate to the pancreatic duct. The 135 SAP rat models after being prepared were randomly divided into the model group, Baicalin treatment group and Octreotide treatment group with 45 rats in each group; another 45 were selected to be the sham operation group, which only received abdomen opening surgery. The groups were then randomly divided into 3 h, 6 h and 12 h groups with 15 rats in each group, 10 min after successful modeling, the Baicalin treatment group was first injected with a 5% Baicalin injection at a dose of 10 mg/100 g via external jugular-vein passage followed by continuous intravenous administration (10 mg/h/100 g) by microinfusion pump; the Octreotide treatment group was first injected by Octreotide at a dose of 0.2 ug/100 g via external jugular-vein passage followed by continuous intravenous transfusion (10 mg/h/100 g) by microinfusion pump at a transfusion speed of 0.2 ug/h/100 g. The sham operation group and model group were injected with saline of equivalent volume at the corresponding time points after operation. The following observations were carried out 3, 6 and 12 h after operation: (1) mortalities of all rat groups followed by batch execution of rats and observation of the gross pathological changes of multiple organs; (2) observation of the pathological changes of multiple organ samples fixed according to the relevant requirements after HE staining; and (3) serum content of amylase, NO, malonaldehyde (MDA), and tumor necrosis factor alpha (TNF-α). Results (1) The survival rate of the sham operation group and all treatment groups was 100%, whilst the 12 h survival of the model group was 66.67% (10/15), indicating a significant difference (P < 0.05). (2) The gross pathological changes and changes under light microscopy of multiple organs aggravated with time after modeling. The pathological changes of all treatment groups were milder than those of the model group at different time points by various degrees, most obviously at 6 h and 12 h. The gross pathological changes showed a similarity between the Octreotide and Baicalin treatment groups in terms of the pathological changes of pancreatic tissue. The therapeutic effects of Octreotide on kidney and lung were superior to those in the Baicalin treatment group while the pathological manifestations of the Baicalin treatment group were superior to those of the Octreotide treatment group. (3) There was no marked difference between the Baicalin and Octreotide treatment groups in terms of plasma amylase levels at all time points (P > 0.05). Although the plasma amylase levels of the Baicalin treatment group were lower than those of the model group at all time points, the levels in the Baicalin treatment group were significantly lower than those in the model group only at 3 h (P < 0.05), and there was no marked difference in the levels between the Baicalin treatment group and model groups at 6 and 12 h (P > 0.05); the levels in the Octreotide treatment group were significantly lower than in the model group at 6 h (P < 0.05), and there was no marked difference between the levels in the Octreotide treatment group and model group at 3 h and 12 h (P > 0.05). (4) The serum NO contents of the Baicalin treatment group were significantly lower than those of the model group (P < 0.05), while in the Octreotide treatment group it was obviously lower than in the model group at 3 and 12 h (P < 0.01); in this regard there was no marked difference between the Baicalin and Octreotide treatment groups at different time points (P > 0.05). (5) The serum MDA contents of the Baicalin treatment group were significantly lower than those of the model group (P < 0.01), while in the Octreotide treatment group it was significantly less than the model group at 6 and 12 h (P < 0.05), and in the Baicalin treatment group was significantly less than in the Octreotide treatment group at 12 h (P < 0.05). (6) There was no marked difference among the model group, Baicalin treatment group and Octreotide treatment group in terms of serum TNF-α content at 3 h and 12 h (P > 0.05). At 6 h the value in the Baicalin treatment group was significantly less than in the model group (P < 0.001), in the Octreotide treatment group it was significantly less than in the model group (P < 0.001), and the Octreotide treatment group it was significantly less than in the Baicalin treatment group (P < 0.01). Conclusions Both Baicalin and Octreotide have obvious protective effects on the multiple organ injury in SAP with mechanisms associated to manifold factors. By comparing the pharmacologic effects of Octreotide and Baicalin, we believe that Baicalin as a new drug has a protective effect on multiple organs of a SAP rat model similar to that of Octreotide and is worth further study and development. This work was supported by the technological foundation project of Traditional Chinese Medicine Science of Zhejiang province (nos. 2003C130 and 2004C142), the foundation project for medical science and technology of Zhejiang province (no. 2003B134), the grave foundation project for technological and development of Hangzhou (No. 2003123B19), intensive foundation project for technology of Hangzhou (no. 2004Z006), the foundation project for medical science and technology of Hangzhou (no. 2003A004), and the foundation project for technology of Hangzhou (No. 2005224). Note: We claim that the article is original and will not have any financial interest in a company or its competitor, and that all authors meet the criteria for authorship.     

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.     

Hepatic effect of NAC on sevear acute pancteatise of rats

Objective:To analyze the hepatic protection of n-acetvl cysteine(NAC)on severe acute pancreatitis(SAP).Methods:SD rats were randomly divided into control group,SAP group and NAC group.SAP AHO method was adopted to establish the model,2 h after modeling,rats in NAC group had intraperitoneal injection of NAC(200 mg/kg).Ten rals from each group were sacrifieed in even'6 and 12 h at different time[mints respectively.Liver damage,liver function and serum amylase,AST,ALT and malondialdehyde(MDA)were determined.Results:Serum amylase,AST,AIT and MDA content in SAP,NAC group at each time point were significantly higher in the control group(P0.05),serum amylase,AST,ALT and MDA content in NAC group rats were lower in the SAP group significantly(P0.05);Microscopic examination showed that the liver injury in rats and the NAC group significantly reduced in the SAP group.Conclusions:NAC provides effective protection against liver damage to SAP,protective from SAP liver injury.     

黄芩甙对重症急性胰腺炎大鼠胰腺氧化应激的保护作用

目的探讨黄芩甙对大鼠重症急性胰腺炎（SAP）氧化应激的保护作用。方法45只大鼠随机分为假手术组、模型组和黄芩甙干预组,每组15只。假手术组进腹后仅翻动胰腺和十二指肠后关腹,不给予任何干预,模型组、黄芩甙干预组采用3.5%牛磺胆酸钠逆行胰胆管注射制造SAP大鼠模型。黄芩甙干预组在造模成功后给予黄芩甙治疗,其余两组给予等量生理盐水。术后3h、6 h及12 h,测定各组大鼠血清淀粉酶的水平及胰腺组织MDA、SOD及MPO的含量,免疫组化法检测胰腺组织聚腺苷二磷酸核糖（PAR）及硝基酪氨酸的表达。结果术后3 h、6 h及12 h,模型组血清淀粉酶较假手术组显著增高（P〈0.01）,黄芩甙干预组血清淀粉酶显著降低于模型组（P〈0.01）;术后各时间点,黄芩甙干预组胰腺组织MDA、MPO含量显著低于模型组（P〈0.01）;SOD水平显著高于模型组（P〈0.01）;术后各时间点,假手术组胰腺组织均无PAR及硝基酪氨酸表达,模型组PAR及硝基酪氨酸表达阳性,且随时间延长表达增多,黄芩甙干预组PAR及硝基酪氨酸表达均低于模型组,且随时间延长显著减少。结论黄芩甙可能通过降低胰腺炎的氧化应激水平,减轻胰腺炎症程度而发挥对胰腺炎的保护作用。     

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.     

Significance of platelet activating factor receptor expression in pancreatic tissues of rats with severe acute pancreatitis and effects of BN52021

AIM:To investigate the dynamic changes and significance of platelet activating factor receptor (PAF-R) mRNA and protein in pancreatic tissues of rats with severe acute pancreatitis (SAP) and effects of BN52021 (Ginkgolide B). METHODS:Wistar male rats were randomly assigned to the negative control group (NC group),SAP model group (SAP group),and BN52051-remedy group (BN group),and each of the groups was 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 in each). PT-PCR and Western blot methods were used to detect PAF-RmRNA and protein expression in pancreatic tissues of rats respectively. Pathological examination of pancreatic tissues was performed and the serum amylase change was detected. RESULTS:Serum amylase and pathological results showed the that SAP model was successfully prepared,BN52021 was able to decrease serum amylase,and the pathological ratings in BN group at 3 h,6 h,and 12 h significantly decreased compared with those in the SAP group (8.85 ± 0.39 vs 5.95 ± 0.19,9.15 ± 0.55 vs 5.55 ± 0.36,10.10 ± 0.65 vs 6.72 ± 0.30,P < 0.05). The result of PAF-mRNA showed dynamic changes in SAP and BN groups,which increased gradually in early stage,reached a peak at 3 h (0.71 ± 0.14 vs 0.54 ± 0.14,0.69 ± 0.13 vs 0.59 ± 0.04,P < 0.05),and decreased gradually later. There were significant differences at each time point except 1 h and 2 h,when compared with those in the NC group (0.71 ± 0.14 or 0.69 ± 0.13 vs 0.47 ± 0.10,0.38 ± 0.08 or 0.59 ± 0.04 vs 0.47 ± 0.09,0.25 ± 0.07 or 0.29 ± 0.05 vs 0.46 ± 0.10,0.20 ± 0.06 or 0.20± 0.04 vs 0.43 ± 0.09,P < 0.05),whereas there was no significant difference between BN and SAP groups at each time point. The result of PAF-R protein showed that the change of PAF-R protein in the SAP group and the BN group was consistent with that of PAF-R mRNA. There were significant differences at each time point except 1 h,when compared with those in the NC group (0.90 ± 0.02 or 0.80 ± 0.05 vs 0.48 ± 0.02,1.69 ± 0.06 or 1.58 ± 0.02 vs 0.48 ± 0.03,1.12 ± 0.10 or 0.98 ± 0.03 vs 0.49 ± 0.09,1.04 ± 0.14 or 0.87 ± 0.02 vs 0.52 ± 0.08,0.97 ± 0.16 or 0.90 ± 0.05 vs 0.49 ± 0.10,P < 0.05),whereas there was no significant difference between the BN group and the SAP group. CONCLUSION:PAF-R plays an important role in occurrence and development of SAP. BN52021 exerts biological effects through competitively inhibiting the binding of increased both PAF and PAF-R expression rather than through decreasing PAF-R expression in pancreatic tissues.