内源性一氧化氮对大鼠急性坏死性胰腺炎的胰腺保护作用

目的 探讨内源性一氧化氮（ＮＯ）对大鼠实验性急性坏死性胰腺炎的作用及其与胰腺血流的关系。方法 经胰胆管内注射５％牛磺胆酸钠溶液（１ｍｌ／ｋｇ）制成Ｗｉｓｔａｒ大鼠急性坏死性胰腺炎模型。以Ｌ－硝基精氨酸（Ｌ－ＮＮＡ）为内源性ＮＯ的阻断剂,观察内源性ＮＯ对胰腺损伤程度的影响;不同时相胰腺炎大鼠胰腺血流量的改变;内源性ＮＯ对损伤状态下大鼠胰腺血流量的影响。结果 经胰胆管注射牛磺胆酸钠可造成大鼠胰腺组织明     

一氧化氮胰腺保护作用与巯基物质和氧自由基的关系

目的　探讨内源性一氧化氮 (NO)对大鼠急性坏死性胰腺炎的作用及其与巯基物质和脂质过氧化之间的关系。　方法　以 5 %牛磺胆酸钠溶液胰胆管注射 (1ml/kg)制成大鼠急性坏死性胰腺炎模型 ,以工具药L 硝基精氨酸 (L NNA)为内源性NO的阻断剂 ,观察内源性NO对胰腺损伤程度、血清淀粉酶浓度、胰腺组织内巯基物质含量和脂质过氧化终产物丙二醛 (MDA)含量的影响。　结果　牛磺胆酸钠胰胆管注射可造成胰腺组织明显的水肿和坏死 ,部分 (2 / 7)发生胰腺实质内出血 ;血清淀粉酶浓度显著升高 ,胰腺组织巯基物质含量降低 ,MDA含量增加 [(1 2 5± 0 2 8)nmol/mg蛋白质vs.(0 5± 0 0 3)nmol/mg蛋白质 ,P <0 0 5 ]。以L NNA(12 5mg/kg)阻断内源性NO ,可明显加重胰腺组织坏死 ,胰腺实质内出血率增加 (10 / 12 ,83 3 % ) ,并血清淀粉酶浓度进一步升高 ,胰腺组织MDA含量进一步增加 [(3 0± 0 40 )nmol/mg蛋白质vs.(1 2 5± 0 2 8)nmol/mg蛋白质 ,P <0 0 5 ]。但对胰腺组织内巯基物质的含量没有影响。　结论　内源性NO具有胰腺保护作用 ,其保护机制可能与抗氧自由基有关。巯基物质可能不参与NO的胰腺保护机制。     

牛磺胆酸钠混合大肠杆菌逆行胰管注射法建立感染性坏死性胰腺炎大鼠模型

目的建立一个稳定可靠的感染性坏死性胰腺炎大鼠模型,为深入研究感染性坏死性胰腺炎的病理生理、病变转归以及探索新的治疗方法提供动物模型载体.方法46只SD大鼠随机分成5组：牛磺胆酸钠逆行胰管注射组、大肠杆菌逆行胰管注射组,牛磺胆酸钠混合不同浓度大肠杆菌（浓度分别为103个/ml、104个/ml和105个/ml,混合液为实验即时配制）逆行胰管注射组,注射量为0.1 ml/100 g体重,注射速度为0.2 ml/min.观察8 h,记录生存率;存活者8 h后活杀,抽血测定血清淀粉酶,并取胰腺组织行病理学检查和细菌培养.结果单纯牛磺胆酸钠逆行胰管注射能建立坏死性胰腺炎模型,其中胰腺组织细菌培养阳性率为12.5%（1/8）;单纯大肠杆菌逆行胰管注射不仅不能建立坏死性胰腺炎模型,而且胰腺组织细菌培养阳性率为0（0/8）;牛磺胆酸钠混合大肠杆菌逆行胰管注射能够建立感染性坏死性胰腺炎模型,大肠杆菌浓度为103个/ml组、104个/ml组、105个/ml组胰腺组织细菌培养率分别为60%、100%、100%,而8 h存活率分别为100%、100%、70%.结论（1）大肠杆菌浓度为104个/ml和牛磺胆酸钠浓度为5%的混合液,按0.1 ml/100 g体重的量经胰管逆行注射（注射速度为0.2 ml/min）可建立稳定可靠的感染性坏死性胰腺炎大鼠模型.（2）该方法导致感染性坏死性胰腺炎发生的可能机制为：牛磺胆酸钠导致胰腺组织发生出血、坏死,并引起胰腺组织抵抗细菌定植能力下降;同时,坏死的胰腺组织给细菌提供了良好的生长环境;此外,胰腺组织发生细菌感染与入侵的细菌量有正相关关系.     

蛋白酶激活受体-2在重症急性胰腺炎中的作用

目的 探讨蛋白酶激活受体-2(PAR-2)在大鼠重症急性胰腺炎病理损害中发挥的作用.方法 由大鼠胰胆管逆行注射3.5%牛磺胆酸钠,制作坏死性胰腺炎模型.36只大鼠被随机分为对照组、胰腺炎组、胰腺炎+皮下活化肽组(给药组),造模后3 h取材.免疫组化方法测定造模前后胰腺组织PAR-2表达情况变化;胰腺损害程度采用病理评分评价;测定胰腺组织湿/干重比、髓过氧化物酶含量,通过股静脉注射Evans blue(EB)测定EB漏出率;ELISIA方法测定血清IL-6水平.结果 正常胰腺组织胞浆及胞膜弱表达PAR-2,造模后表达升高.给药组病理评分、湿/干重比、髓过氧化物酶含量、Evans blue漏出率、血清IL-6水平明显低于胰腺炎组,胰腺病理评分与IL-6之间存在相关关系.结论 诱导重症急性胰腺炎后胰腺PAR-2表达升高,PAR-2可明显减轻胰腺炎症程度,对胰腺局部损害起保护作用.     

前列腺素E1防治大鼠急性坏死性胰腺炎的作用及机理

目的：研究前列腺素Ｅ１防治大鼠急性坏死性胰腺炎的作用及其机理。方法：向大鼠胰管内注射５％的牛磺胆酸钠溶液制成急性坏死性胰腺炎（ＡＮＰ）模型。结果：ＡＮＰ时，且腺毛细血管通透性（ＰＣＰ）明显增高，胰腺组织中性粒细胞过氧化酶（ＭＰＯ）活性及脂质过氧化物（ＬＰＯ）水平明显增高。病理示组织内大量ＰＭＮ浸润、片状出血、坏死。ＰＧＥ１使ＰＣＰ明显下降，ＭＰＯ、ＬＰＯ显著降低，动物存活明显改善结论：ＰＧＥ１通过     

急性出血坏死性胰腺炎大鼠胰腺组织结构改变与内毒？…

为探讨大鼠急性出血坏死性胰腺炎（ＡＨＮＰ）胰腺组织结构改变与内毒素血症的关系及纳屈桐的治疗作用，用５％牛磺胆酸钠逆行胰胆管注射制成ＡＨＮＰ模型。取Ｗｉｓｔａｒ大鼠１１０只随机分为假手术组（ｎ＝２０）ＡＨＮＰ组，纳屈酮治疗组，分别手术后６、１２、２４小时称取胰腺重量，观察ＡＨＮＰ大鼠胰腺组织结构改变。同时测定其血浆淀粉酶和内毒素水平，并与假手术组比较。结果：与假手术组比较，ＡＨＮＰ组血浆淀粉酶、内毒     

小剂量多巴胺对急性出血坏死性胰腺炎大鼠胰腺微血流的影响及其治疗作用

应用小剂量多巴胺[5μg／（kg·min）]对5％牛磺胆酸钠诱导的大鼠急性出血坏死性胰腺炎（AHNP）模型进行治疗，以观察其对AHNP大鼠平均动脉压、胰腺微区血流量、血清淀粉酶和脂肪酶的影响及胰腺的病理学改变。结果：AHNP早期在不影响平均动脉压的情况下，小剂量多巴胺能显著增加大鼠胰腺微区血流量，降低血清淀粉酶和脂肪酶，并减轻胰腺病理改变程度。实验结果提示：小剂量多巴胺在改善胰腺微血流的基础上对胰腺炎有治疗作用。     

前列腺素E_1防治大鼠急性坏死性胰腺炎的作用及机理

研究前列腺素E_1防治大鼠急性坏死性胰腺炎的作用及其机理。方法:向大鼠胰管内注射5％牛磺胆酸钠溶液制成急性坏死性胰腺炎(ANP)模型。结果:ANP时,胰腺毛细血管通透性(PCP)明显增高,胰腺组织中性粒细胞过氧化酶(MPO)活性及脂质过氧化物(LPO)水平明显增高。病理示组织内大量PMN浸润、片状出血、坏死。PGE_1使PCP明显下降,MPO、LPO显著降低,动物存活明显改善结论:PGE_1通过抑制中性粒细胞(PMN)活化减少氧自由基(OFR)释放,减轻血管内皮细胞及胰腺组织损伤。     

炎症介质与急性坏死性胰腺炎大鼠的肺损伤

目的　探讨急性坏死性胰腺炎（ANP）大鼠并发肺损伤的机制。方法　逆行性胰胆管注射3.5%牛磺胆酸钠建立ANP大鼠模型。观察肺组织学改变,测定血中淀粉酶、磷脂酶A2（PLA     

小剂量多巴胺对急性出血坏死性胰腺炎大鼠胰腺微血流的…

应用小剂量多巴胺（５μｇ／（ｋｇ．ｍｉｎ）对５％牛磺胆酸钠诱导的大鼠急性出血坏死性胰腺炎（ＡＨＮＰ）模型进行治疗，以观察其对ＡＨＮＰ大鼠平均动脉压，胰腺微区血流量，血清淀粉酶和脂肪酶的影响及胰腺的病理学改变，结果：ＡＨＮＰ早期在不影响平均动脉压的情况下，小剂量多巴胺能显著增加大鼠胰腺微区血流量，降低血清淀粉酶和脂肪酶，并减轻胰腺病理改变程度，实验结果提示，小剂量多巴胺在改善胰腺管微血流的基础上对胰     

N-乙酰半胱氨酸对大鼠急性坏死性胰腺炎胰腺损伤的影响

目的 探讨急性坏死性胰腺炎(ANP)胰腺损伤与核因子-κB(NF-κB)活化、胰腺细胞凋亡的关系及N-乙酰半胱氨酸(NAC)对胰腺损伤的影响.方法 33只Wistar大鼠分为正常对照、盐水对照和胰腺炎组.以3.5%牛磺胆酸钠逆行注入胰胆管制作ANP模型,于造模前、后1 h应用NAC,12 h后取材,采用凝胶电泳迁移率实验测定胰腺组织NF-κB活性、改良TUNEL法检测细胞凋亡.同时观察血淀粉酶、脂肪酶、胰腺组织湿/干重比率及病理改变. 结果盐水对照组NF-κB活性很低(2.00±0.33),胰腺炎组NF-κB明显活化(6.03±0.41),造模前使用NAC抑制NF-κB活性(3.28±0.42),降低淀粉酶及胰腺湿/干重比率,促进胰腺细胞凋亡(P<0.05).NF-κB活化与凋亡呈负相关(r=-0.96,P<0.01),与胰腺损伤病理呈正相关(r=0.63,P<0.01);胰腺损伤病理分级与凋亡呈负相关(r=-0.98,P<0.01).结论 NAC可能通过抑制胰腺NF-κB活化、促进胰腺细胞凋亡,减轻胰腺损伤.     

ET-1 induces pancreatitis-like microvascular deterioration and acinar cell injury.

Using in vivo microscopy red blood cell (RBC) velocities, functional capillary density (FCD) and capillary diameters were estimated after inducing acute pancreatitis by intraductal infusion of sodium taurocholate (0.8 ml; 4%) or after topical superfusion of the pancreas with ET-1 (100 pmol). Sodium taurocholate mediated a significant decrease in RBC velocities between 50 and 70%, transient decrease in capillary diameters by 10%, and a sustained decrease in FCD between 60 and 70% paralleled by a dramatic heterogeneity in blood flow. Topical superfusion of the exteriorized pancreas with ET-1 caused a significant decrease in RBC velocities between 65 and 75%, a sustained decrease in capillary diameters by 10%, and a decrease in FCD by 45% accompanied by an increase in flow heterogeneity. Following sodium taurocholate infusion pancreas histology revealed a severe edema and sublobular acinar cell necrosis, while topical ET-1 application displayed a severe edema of the pancreas with focal acinar cell necrosis. Thus, ET-1 mediated a deterioration of the pancreatic microcirculation, which is similar to the microcirculatory failure found in sodium taurocholate-induced experimental pancreatitis and was associated with focal acinar cell necrosis. We are thus inclined to hypothesize that endothelin released by injured endothelial cells during acute biliary pancreatitis promotes microcirculatory failure and ischemia in acute pancreatitis, eventually leading to acinar cell necrosis.     

急性胰腺炎大鼠TNF-α mRNA、IL-10 mRNA表达和胰腺细胞凋亡的研究

目的 探讨大鼠急性胰腺炎早期胰腺组织中TNF－αmRNA、IL－10mRNA的表达和细胞凋亡的变化规律。方法 以牛磺胆酸钠诱导20只大鼠急性水肿性胰腺炎（AEP）模型,20只急性坏死性胰腺炎（ANP）模型,另取10只正常大鼠作为对照。术后12h各处死10只大鼠,检测血清和胰腺组织中的TNF－α和IL－10水平,分析两者在胰腺组织中的mRNA较录水平,检测胰腺细胞的凋亡率。结果 正常、AEP和ANP组的细胞凋亡率分别为2．98％、17．29％和8．39％。制模后TNF－αm和IL－10增强ANP大鼠TNF－α表达增强。结论 急性胰腺炎大鼠胰腺组织中的TNF－α和IL－10的表达与其在血清和胰腺中的浓度成正比,胰腺本身可能就是产生细胞因子的主要器官。胰腺细胞凋亡率与疾病的严重程度呈负相关,凋亡是对胰腺损伤的良好反应。     

早期应用L-精氨酸治疗急性出血坏死性胰腺炎的实验研究

目的: 探讨早期应用L-精氨酸对急性出血坏死性胰腺炎(AHNP)的治疗作用. 方法: 经大鼠胰胆管注射牛磺酸钠制备AHNP模型,分别静注L-精氨酸、生理盐水、山莨菪碱;观察血清淀粉酶、白介素6(IL-6)、丙二醛(MDA)、一氧化氮(NO)改变及胰腺的病理改变. 结果: 应用L-精氨酸后,能升高血NO浓度,清除氧自由基,降低血清IL-6水平,降低血清淀粉酶,改善胰腺病变,其作用优于山莨菪碱. 结论: 早期应用一定剂量的L-精氨酸对AHNP具有一定的治疗效果.     

水通道蛋白1在急性坏死性胰腺炎大鼠胰腺组织的表达及其意义

目的 探讨水通道蛋白1(AQP1)在急性坏死性胰腺炎(ANP)大鼠胰腺的表达及其意义.方法 将48只雄性SD大鼠随机分为对照组和ANP组,制模后3、6、12、18 h各时间点分别处死6只.记录腹水量,测定血清淀粉酶;采用酶联免疫吸附试验(ELISA)检测血清AQP1含量;苏木素-伊红(HE)染色观察胰腺组织病理改变;伊文思兰染料(EB)血管外渗法检测胰腺组织毛细血管通透性;免疫组织化学和Westem blot法检测胰腺组织AQP1蛋白表达;荧光定量聚合酶链反应(PCR)检测AQP1基因mRNA表达.结果 (1)对照组3、6、12、18 h血清淀粉酶水平分别为(1308±759)、(1077±508)、(1325±761)、(1328±762)U/L,ANP组分别为(9102±2199)、(8799±1634)、(9398±1473)、(9484±862)U/L;对照组胰腺组织EB含量分别为(205.61±32.99)、(141.46±27.18)、(96.94±26.79)、(61.43±24.82)mg/L;ANP组分别为(273.59±23.47)、(253.51±31.68)、(221.15±73.68)、(185.28±42.35)mg/L;血清AQP1含量对照组为(74.08±11.80)、(78.49±9.06)、(75.77±7.37)、(72.75±13.87)mg/L,ANP组为(73.29±9.61)、(62.85±7.28)、(62.07±4.39)、(46.33±11.91)mg/L,两组比较差异均有统计学意义(P<0.01).(2)对照组3、6、12、18 h免疫组织化学灰度值分别为114.13±7.92、122.39±7.99、145.98±6.48、113.98±6.48,ANP组分别为80.07±14.89、110.54±4.45、103.77±10.48、99.18±6.95;对照组Western blot蛋白含量分别为1.19±0.33、1.02±0.25、0.90±0.33、1.06±0.20,ANP组分别为0.83±0.11、0.96±0.21、0.58±0.28、0.72±0.14.结果 均显示ANP组胰腺AQP1蛋白表达低于对照组(P<0.05);(3)对照组3、6、12、18 h荧光定量PCR检测ANP组胰腺AQP1基因mR-NA表达分别为2.13±0.63、2.02±1.40、2.07±0.86、2.49±2.47,ANP组为0.91±0.22、1.01±0.83、0.48±0.23、0.61±0.51,ANP组较对照组减弱(P<0.01).结论 ANP大鼠胰腺组织AQP1表达明显减弱,这可能在毛细血管渗漏综合征的发生中起重要作用.     

一种研究急性胰腺炎加重病理机理的动物模型

介绍一种急性水肿性胰腺炎（ＡＥＰ）向坏死性胰腺炎（ＡＮＰ）转变的大鼠模型。１０７只ＳＤ大鼠随机分为假手术组、ＡＥＰ组和ＡＮＰ组。ＡＥＰ通过胰管结扎、外分泌刺激诱发。在ＡＥＰ模型基础上静注大剂量Ｄｅｘｔｒａｎ１１０诱发ＡＮＰ。结果显示：血清淀粉酶水平在ＡＥＰ、ＡＮＰ组明显增高，胰腺泡细胞胞浆游离钙离子浓度在ＡＮＰ诱发后持续增高；胰腺出血、实质坏死、钙沉积在ＡＮＰ组常见。超微结构显示ＡＮＰ组胰毛细血管内皮剥脱、坏死。由此表明，胰腺缺血可能通过腺泡细胞钙超负荷的作用促发ＡＥＰ向ＡＮＰ转变。该大鼠模型因临床联系较好、病变渐进，不失为一种从细胞和分子水平研究急性胰腺炎加重病理机理的动物模型     

Connective Tissue Growth Factor is Involved in Pancreatic Repair and Tissue Remodeling in Human and Rat Acute Necrotizing Pancreatitis

OBJECTIVE: To analyze the involvement of connective tissue growth factor (CTGF) in the transforming growth factor-beta (TGF-beta) pathway during acute necrotizing pancreatitis (ANP) in humans and rats. SUMMARY BACKGROUND DATA: Connective tissue growth factor is involved in several fibrotic diseases and has a critical role in fibrogenesis and tissue remodeling after injury. METHODS: Normal human pancreas tissue samples were obtained through an organ donor program from five individuals without a history of pancreatic disease. Human ANP tissues were obtained from eight persons undergoing surgery for this disease. In rats, ANP was induced by intraductal infusion of taurocholate. The expression of CTGF was studied by Northern blot analysis, in situ hybridization, and immunohistochemistry in both human and rat pancreatic tissue samples. RESULTS: Northern blot analysis revealed enhanced CTGF mRNA expression in human ANP tissue samples compared with normal controls. In addition, a concomitant increase in TGF-beta1 was present. By in situ hybridization, CTGF mRNA was localized in the remaining acinar and ductal cells and in fibroblasts. In regions of intense damage adjacent to areas of necrosis, CTGF mRNA signals were most intense. Inflammatory cells were devoid of any CTGF mRNA signals. By immunohistochemistry, CTGF protein was localized at high levels in the same cell types as CTGF mRNA. In ANP in rats, concomitantly enhanced mRNA levels of CTGF, TGF-beta1, and collagen type 1 were present, with a biphasic peak pattern on days 2 to 3 and day 7 after induction of ANP. CONCLUSIONS: These data indicate that CTGF participates in tissue remodeling in ANP. The expression of CTGF predominantly in the remaining acinar and ductal cells indicates that extracellular matrix synthesis after necrosis is at least partly regulated by the remaining pancreatic parenchyma and only to a minor extent by inflammatory cells. Blockage of CTGF, a downstream mediator of TGF-beta in fibrogenesis, might be useful as a target to influence and reduce fibrogenesis in this disorder.     

Inhibition of matrix metalloproteinases reduces local and distant organ injury following experimental acute pancreatitis

BACKGROUND: Pulmonary complications from pancreatitis involve parenchymal destruction via proteolytic enzymes. Matrix metalloproteinases (MMPs) may play an important role in pulmonary injury following acute severe pancreatitis. We hypothesized that local and distant organ injury would be decreased by the presence of an MMP inhibitor (Batimistat; BB-94) following severe acute pancreatitis (AP). METHODS: Eighteen male rats were randomized into two groups: BB-94 (AP + 40 mg/kg/24 h BB-94 ip x three doses) or control (AP + 20 ml/kg/24 h normal saline ip x three doses). Necrotizing AP was induced by retrograde infusion of 5% sodium taurocholate (1.5 ml/kg) into the pancreatic duct. Twenty additional animals were randomized into BB-94 and control groups for the survival study. Serum was evaluated for amylase and MMP activity. Pancreatic sections were graded for edema, necrosis, neutrophil infiltrate, and hemorrhage. Myloperoxidase (MPO) activity was used to determine PMN infiltration in the lung. Evan's Blue dye extravasation was used to quantify vascular permeability. RESULTS: Animals in the BB-94 group had decreased amylase levels (1086.0 +/- 61.7 U/L vs 2232.7 +/- 309.9 U/L; P < 0.05), decreased cellular infiltrate (1.4 +/- 0.2 vs 2.3 +/- 0.2; P < 0.02), and decreased necrosis (4.1 +/- 0.3 vs 6.1 +/- 0.4; P < 0.005) compared to the control group. Lung tissue following pancreatitis in the BB-94 group demonstrated decreased MPO activity (41.5 +/- 2.4 units vs 57.3 +/- 2.9 units; P < 0.05) and decreased vascular permeability (18.3 +/- 2.8 mg/100 g vs 30.1 +/- 4.6 mg/100 g; P < 0.05). Animals treated with BB-94 had 100% survival compared to 50% survival in control at 72 h. CONCLUSIONS: Pancreatitis results in increased local and distant MMP activity. Pulmonary and pancreatic injury following AP can be abrogated by treatment with an MMP inhibitor (Batimistat; BB-94) which may result in decreased morbidity and mortality.