高迁移率族蛋白B1与大鼠急性胰腺炎模型病情严重程度的关系

目的探讨高迁移率族蛋白B1(HMGB1)与大鼠急性胰腺炎模型病情严重程度关系.方法制作大鼠重症急性胰腺炎A组、轻症急性胰腺炎B组、正常对照组C组模型各10只,于造模后12 h、24 h、48 h采血,采用酶联免疫法(ELISA)检测各组大鼠模型不同时间段血清HMGB1含量的变化.结果大鼠血清HMGB1在各时间点测量值A组高于B组高于C组,P<0.05,具有统计学意义.结论大鼠血清HNGB1与急性胰腺炎的病情严重程度成正相关.

大黄牡丹汤组方对急性胰腺炎模型大鼠胰腺微循环的影响

目的研究大黄牡丹汤组方(RPDP)对急性胰腺炎(AP)模型大鼠胰腺早期胰腺微循环的改变,探讨微循环改变在AP发生发展中的作用,观察RPDP对胰腺微循环的影响。方法 60只SD大鼠随机分为药物作用4 h假手术(SO)组、模型组和RPDP组;药物作用8 h SO组、模型组和RPDP组,每组各10只。模型组和RPDP组在距离胆胰管开口0.3 mm处的胆管,经十二指肠壁浆肌层针刺,用3.5%牛磺胆酸钠0.1 m L/100 g缓慢注射,建立SD大鼠AP模型,治疗组用RPDP 40 g/kg灌胃1次。分别与给药后4 h和8 h,麻醉相应组大鼠抽取血液及腹水,检测各组血清和腹水淀粉酶含量。用荧光显微流速测试方法,观察胰腺微血管在造模后4 h和8 h的变化。结果模型组大鼠胰腺微循环血流速度存在差异,RPDP治疗组血流速度则比较一致。与SO组比较,模型组大鼠血清淀粉酶及腹水淀粉酶均显著升高;与模型组比较,RPDP组大鼠血清淀粉酶及腹水淀粉酶均显著降低(P<0.01)。结论 AP发病与胰腺微循环障碍有关,AP向重症急性胰腺炎(SAP)发展也与其有关,AP时胰腺水肿与坏死同时存在。RPDP有改善AP模型大鼠胰腺微循环的作用。

小鼠胰腺炎模型中胰腺导管腺体与导管上皮修复再生的研究

目的:探讨在炎症情况下胰腺导管腺体(PDG)与胰腺导管上皮修复再生过程的关系。方法:采用病理学评分评估雨蛙肽诱导急慢性胰腺炎小鼠模型的建立,采用病理组织学方法观察胰腺组织中PDG及导管结构病理学变化;免疫组化检测PDG及胰腺导管上皮细胞中Pdx1、Sox9、Ngn3的表达情况,利用免疫组化评分(HIS评分)评估其阳性表达率。结果:成功建立小鼠急慢性胰腺炎模型,其病理学评分分别为3.33±0.52和8.50±0.55,与对照组相比差异有统计学意义;在慢性胰腺炎模型中,PDG结构出现明显的增殖变化,且Pdx1、Sox9、Ngn3的表达均有阳性,HIS评分分别为1.966 7±0.233 8、0.700 0±0.209 8、0.900 0±0.209 8,与对照组相比差异有统计学意义。结论:PDG结构与胰腺导管上皮组织的修复再生相关,且与成体胰腺的分化成熟有联系,可能以胰腺干细胞巢的形式分布于胰腺组织中。

DDC诱导大鼠慢性胰腺炎动物模型的探讨

目的：用DDC大鼠腹腔注射法，观察胰腺的病理学变化和TGF—β1的表达，为建立慢性胰腺炎（CP）的动物模型做可行性探讨。方法：用DDC对实验（model，M）组大鼠腹腔注射，对照组（contrast，c）用生理盐水代替DDC进行相同的操作。结果：实验组大鼠有明显慢性胰腺炎的病理改变；对照组胰腺组织呈现正常状态。同时实验组TGF—β1表达比对照组增高，有显著差异（P〈0．05）。结论：DDC可在较短的时间诱导大鼠慢性胰腺炎，导致胰腺纤维化，并引起TGF—β1高表达。

丹参酮ⅡA磺酸钠对大鼠急性胰腺炎高迁移率族蛋白B1下调作用的研究

目的探讨丹参酮ⅡA磺酸钠注射液是否对大鼠急性胰腺炎模型血清高迁移率族蛋白B1(HMGB1)有下调作用.方法复制重症急性胰腺炎组(A1组)及重症急性胰腺炎治疗组(A2组),轻症急性胰腺炎组(B1组)及轻症急性胰腺炎治疗组(B2组)动物模型,每组10只大鼠,A2、B2组于造模后3、27 h在大鼠后大腿外侧肌注丹参酮IIA磺酸钠注射液(4 mg/kg).于造模后12 h、24 h、48 h采血,采用酶联免疫法(ELISA)检测各组大鼠模型不同时间段血清HMGB1含量的变化.结果血清HMGB1测量值在各时间点A1组均高于A2组(P<0.05),B1组均高于B2组(P<0.05),差异具有统计学意义.结论丹参酮IIA磺酸钠注射液对大鼠急性胰腺炎模型血清HMGB1有拮抗作用.

TNF-α、IL-10在实验性重症急性胰腺炎中的表达

目的通过测定TNF-α、IL-10在实验性重症急性胰腺炎中的表达,初步探讨二者在重症急性胰腺炎中的作用。方法用逆行性胰胆管注射牛磺胆酸钠方法制备大鼠重症胰腺炎模型。检测模型血浆TNF-α、IL-10水平并与对照组作比较研究。结果 SAP组各时点血浆TNF-α含量与假手术组比较,存在显著性差异(P<0.05);SAP组各时点之间两两比较,存在显著性差异(P<0.05);SAP组各时点血浆IL-10含量与假手术组比较,存在显著性差异(P<0.05);SAP组各时点之间两两比较,存在显著性差异(P<0.05)。结论 TNF-α、IL-10与重症急性胰腺炎密切相关。*

胆源性胰腺炎发病机理探讨

本文采用85只大白鼠制成胆汁返流性胰腺炎模型,并总结139例胆源性胰腺炎患者的临床资料,藉以探讨其发病机理。作者认为:急性胰腺炎与胆道疾病关系甚为密切。

氧自由基和血浆内毒素含量在犬急性胰腺炎病理模型中的变化研究

本试验选取10只本地健康杂交犬,随机分为空白对照组和急性胰腺炎组,每组5只。急性胰腺炎组采用胰管逆行注射法即在主胰管注入牛磺胆酸钠与胰蛋白酶复合液构建急性胰腺炎模型。然后分别在造模后1d,2d,4d,7d采集静脉血,动态地检测其血浆内毒素(ET)、丙二醛(MDA)、超氧化物歧化酶(SOD)等含量变化。结果显示:(1) 对照组内毒素含量在试验期间变化不明显,基本保持在正常值范围之内,表明对照组基本没有出现细菌移位,血液细菌感染;模型组内毒素含量都在第2天达到峰值,之后逐渐下降,在实验第7天基本接近正常水平,说明在急性胰腺炎发生之后细菌移位最严重的时间应该是第2天。(2) 对照组SOD含量在试验期间变化不明显;模型组SOD含量在第1天时变化不明显,等到第4到7天时明显降低(P<0.01),且略微低于对照组。(3) 照组的血浆MDA含量基本稳定在正常范围内,变化幅度不大;模型组的血浆MDA含量在第2天达到峰值,然后逐渐下降,且在各个监测点显著高于对照组(P<0.01)。

microRNAs在急性胰腺炎中的研究新进展

MicroRNAs(miRNAs)是属于小的非编码RNA(small non-coding RNA),长约19~23个核苷酸的内源性非编码单链RNA分子[1-3],约70个碱基大小形成发夹结构的单链RNA前体且经过Dicer酶加工后生成。有5’端磷酸基和3’羟基,大小约21~25nt的小分子RNA片断,定位于RNA前体的3’端或者5’端。目前认为。

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） vs 0,053 （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 h in 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 SAR

Effect of parenteral and early intrajejunal nutrition on pancreatic digestive enzyme synthesis, storage and discharge in dog models of acute pancreatitis

AIM： To study the effect of early intrajejunal nutrition on enzyme-protein synthesis and secretion during acute pancreatitis. METHODS： Fifteen dogs were randomly divided into parenteral nutrition （n = 7） and early intrajejunal nutrition groups （n = 8）. An acute pancreatitis model was induced by injecting 5% sodium taurocholate and trypsin into the pancreas via the pancreatic duct. Intrajejunal nutrition was delivered with a catheter via a jejunostomy tube after the model was established for 24 h. On d 1 and 7 and at the beginning of nutritional support, radioactive tracing and electron microscopes were used to evaluate the enzyme-protein synthesis in acinar cells, the subcellular fractionation and the change in zymogen granules after 1.85 × 10^6 Bq L-3H phenylalanine was infused at 30, 60, 120, and 180 min. RESULTS： The 3H radioactivity in pancreatic acinar cells reached its peak level at 60 min, and the contents in the early intrajejunal nutrition group were higher than those in the parenteral nutrition group, which were then decreased. The mean number and area of zymogen granules did not show any significant statistical difference in both groups on d i or on d 7 （P 〉 0.05）. CONCLUSION： Early intrajejunal nutrition might be effective in dogs with 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.

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)-α, 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 μmol/L vs 38.400 ± 11.344 μmol/L; P < 0.05, 33.533 ± 10.106 μmol/L vs 45.154 ± 17.435 μmol/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) μmol/L vs 65.00 (7.50) μmol/L; P < 0.01, 57.50 (27.50) and 45.00 (12.50) μmol/L vs 74.10 (26.15) μmol/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-α (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.

Evidences supporting the vascular etiology of post-double balloon enteroscopy pancreatitis: Study in porcine model

Double balloon enteroscopy(DBE) is an endoscopic technique broadly used to diagnose and treat small bowel diseases. Among the associated complications of the oral DBE, post-procedure pancreatitis has taken the most attention due to its gravity and the thought that it might be associated to the technique itself and anatomical features of the pancreas. However, as the etiology has not been clarified yet, this paper aims to review the published literature and adds new results from a porcine animal model. Biochemical markers, histological sections and the vascular perfusion of the pancreas were monitored in the pig during DBE practice. A reduced perfusion of the pancreas and bowel, the presence of defined hypoxic areas and disseminated necrotic zones were found in the pancreatic tissue of pigs. All these evidences contribute to support a vascular distress as the most likely etiology of the post-DBE pancreatitis.

A simple taurocholate-induced model of severe acute pancreatitis in rats

AIM: To investigate gut barrier damage and intestinal bacteria translocation in severe acute pancreatitis (SAP), a simple rat model of SAP was induced and studied.METHODS: Pancreatitis was induced by uniformly distributed injection of 3.8% Na taurocholate (1 mL/kg) beneath the pancreatic capsule. Rats in the control group were injected with normal saline in the identical location. RESULTS: Serum amylase, plasma endotoxin, intestinal permeability, and pancreatitis pathology scores were all markedly higher in the pancreatitis group than in the control group (P < 0.01). The bacterial infection rate was signif icantly higher in the SAP group than in the control group (P < 0.01), observed in parallel by both bacterial culture and real-time polymerase chain reaction. Acute damage of the pancreas was observed histologically in SAP rats, showing interstitial edema, leukocyte infiltration, acinar cell necrosis and hemorrhage. The microstructure of the intestinal mucosa of SAP ratsappeared to be destroyed with loose, shortened microvilli and rupture of the intercellular junction, as shown by electron microscopy. CONCLUSION: Significant gut barrier damage and intestinal bacterial translocation were def initely observed with few potential study confounders in this SAP rat model, suggesting that it may be an appropriate animal model for study of gut barrier damage and bacterial translocation in SAP.

Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis

AIM:To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells(MSCs) in severe acute peritonitis(SAP).METHODS:Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups:nonsodium deoxycholate(SDOC) group(non-SODC group),SDOC group,and a MSCs intervention group(i.e.,a co-culture system of MSCs and pancreatic acinar cells + SDOC).The cell survival rate,the concentration of malonaldehyde(MDA),the density of superoxide dismutase(SOD),serum amylase(AMS) secretion rate and lactate dehydrogenase(LDH) leakage rate were detected at various time points.In a separate study,Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group.Serum AMS,MDA and SOD,interleukin(IL)-6,IL-10,and tumor necrosis factor(TNF)-α levels,intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats.In both studies,the protective effect of MSCs was evaluated.RESULTS:In vitro,The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced,and the concentration of MDA,AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point.In vivo,Serum AMS,IL-6,TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group;however serum IL-10 level was higher than the SAP group.Serum SOD level was higher than the SAP group at each time point,whereas a significant betweengroup difference in SOD level was only noted after 24 h.Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs.CONCLUSION:MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium,promote the proliferation of enteric epithelium and repair of the mucosa,attenuate systemic inflammation in rats with SAP.

Effect of dexamethasone,anisodamine and rhubarb therapy on rats with acute pancreatitis

Objective:To investigate the therapeutic effects of dexamethasone,anisodamine and rhubarb(DAR) on endotoxin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and pancreatic damage in rat models of acute pancreatitis(AP).Methods:The AP rat models were prepared and randomly assigned to AP group(n=10) and DAR group(n=10),while other healthy rats were assigned to the sham-operated group(n=10).The rats were euthanized at 6 h after operation,and then the serum levels of endotoxin,TNF-α,IL-6 and histology of pancreas were determined as the indexes of therapeutic effects.Results:At 6 h after operation,serum levels of endotoxin,TNF-α and IL-6,and pancreatic damage were significantly increased in AP group compared with those in sham-operated group(P<0.01).Compared with the AP group,DAR therapy remarkably attenuated the endotoxin,TNF-α,IL-6 levels and reduced pancreatic damage(P<0.05).Conclusion:The inhibition of pancreatic damage by DAR in rats with AP might contribute,in part at least,to the amelioration of pancreatic inflammation.The present study provides beneficial evidence that DAR may be useful in the treatment of AP model of rats.