维拉帕米在高脂血症性胰腺炎大鼠中的研究

发布时间：2018-03-06 09:05

  本文选题：重症急性胰腺炎　切入点：高脂血症性胰腺炎　出处：《泸州医学院》2014年硕士论文　论文类型：学位论文

【摘要】：高脂血症性胰腺炎临床上一般又称作高甘油三酯血症性胰腺炎，随着人们生活水平的提高和饮食结构的改变，其发病率逐渐上升，目前是仅次于胆源性胰腺炎和酒精性胰腺炎的第三大病因，，约占1.3%—3.8%。但高脂血症性胰腺炎的发病机制目前尚无法得到统一的认定，主要集中在游离脂肪酸的毒性作用，钙超载的诱导作用以及基因突变方面的研究。结合新近的论点，本课题针对钙超载诱导机制，做更进一步的研究。 目的：应用钙通道阻滞剂维拉帕米作用于高脂血症性胰腺炎大鼠，通过观察其疗效，探讨钙超载在高脂血症性胰腺炎发病机制中的作用及意义，从而为维拉帕米作为高脂血症性胰腺炎治疗新药物提供理论依据。方法：80只SD雄性大鼠，体重为200-250g，随机分为四组，每组20只，假手术组（SO组）：普通乳剂饲料（普通饲料+适量蒸馏水），对大鼠进行灌胃，持续两周，两周后开腹并用空针穿刺胰胆管，不注射任何物质，然后退针关腹。重症胰腺炎组（SAP组）：普通乳剂饲料灌胃两周的大鼠，开腹后胰胆管逆行缓慢推注5%牛磺胆酸钠，后关腹。高脂血症性胰腺炎组（HLP组）：高脂灌胃剂（20%猪油+10%胆固醇+20%吐温80+2%胆酸钠+1%丙基硫氧嘧啶+适量蒸馏水）灌胃两周后，胰胆管穿刺推注5%牛磺胆酸钠，关腹。维拉帕米治疗组（CCB组）：高脂血症性胰腺炎造模后10分钟，给予腹腔注射维拉帕米1mg/kg。术前经颈静脉或心脏采血检测其甘油三酯（triglyceride，TG）水平；手术24小时后，统计死亡例数，处死所有大鼠，观察并记录腹水量；用酶联免疫吸附法（ELISA法）测定血清血栓素A2（TXA2）及血小板活化因子（PAF）的含量；取适量胰腺组织，用苏木精-伊红染色（HE染色），显微镜下观察其胰腺病理改变，并对胰腺组织损伤进行病理学评分；取适量胰腺组织用流式细胞仪技术检测细胞内钙离子水平；用Western印迹法分析NF-κBp65蛋白的表达。结果：1.SO组大鼠无一死亡，HLP组死亡7只，SAP组死亡4只，CCB组死亡2只。2.SAP组、HLP组、CCB组均有大量血性腹水，而SO组几乎没有腹水；HLP组、CCB组TG水平均较SO组和SAP组显著升高，差异有统计学意义（P＜0.05）。3.胰腺组织损伤病理评分： HLP组、SAP组炎症，水肿，出血，坏死均较SO组显著增高（P0.05）,HLP组损伤水平较SAP组高（P0.05），而CCB组炎症水肿出血坏死水平又明显比HLP组低（P0.05）。4. HLP组与SAP组TXA2、PAF含量均较对照组显著升高（P0.05），HLP组又高于SAP组（P0.05），而CCB组TXA2、PAF含量均低于HLP组，差异有统计学意义（P0.05）。5.钙离子水平： HLP组与SAP组显著高于SO组（P0.05），HLP组钙离子也高于SAP组，差异有统计学意义（P0.05），而CCB组与HLP组比较又显著性降低（P0.05）。6. NF-κBp65蛋白表达： HLP组与SAP组NF-κBp65蛋白表达强度及阳性率均较对照组显著升高（P0.05），而HLP组NF-κBp65蛋白表达强度及阳性率与SAP组、CCB组又有显著性差异（P0.05）。结论：1．用改进型的高脂乳剂（20%猪油+10%胆固醇+20%吐温80+2%胆酸钠+1%丙机硫氧嘧啶+适量蒸馏水）进行灌胃，持续两周，可成功建立高脂血症大鼠模型；胰胆管逆行推注5%牛磺胆酸钠也可成功建立重症急性胰腺炎。2．高脂血症性胰腺炎较普通型胰腺炎大鼠的病情更重，说明高血脂在胰腺炎的发生、发展上起到了重要的推动作用。3．通过运用钙通道阻滞剂维拉帕米抑制胰腺腺泡细胞内钙超载，可适当减轻高脂血症性胰腺炎大鼠的病情程度，说明钙超载参与了高脂血症性胰腺炎的病变过程，也说明应用维拉帕米治疗高脂血症性胰腺炎有效。
[Abstract]:Hyperlipidemic pancreatitis clinical commonly called hypertriglyceridemia pancreatitis, with the improvement of people's living standard and the change of diet structure, the incidence increased gradually, is currently the third leading cause after biliary pancreatitis and alcoholic pancreatitis, accounting for about 1.3% of 3.8%. but the pathogenesis of hyperlipidemic pancreatitis there is no unified identification, mainly focused on the toxic effects of free fatty acids, research and the role of gene mutation induced by calcium overload. Combined with the recent arguments, the topic for the calcium overload induced mechanism, do further research.
Objective: the application of calcium channel blockers Vera Pammy effect on hyperlipidemic pancreatitis rats by observing its efficacy, to investigate the significance and calcium overload in hyperlipidemic pancreatitis pathogenesis, so as to provide a theoretical basis for new drugs for Vera Pammy as hyperlipidemic pancreatitis. Methods: 80 male SD rats. The weight of 200-250g, were randomly divided into four groups, 20 rats in each group, sham operation group (group SO): ordinary emulsion feed (normal diet and proper amount of distilled water), the rats were given orally for two weeks, two weeks after the open and empty needle puncture of pancreatic duct, with no substance, then the needle closed abdomen. Severe acute pancreatitis group (SAP group): ordinary feed emulsion by gavage for two weeks in rats after laparotomy and retrograde cholangiopancreatography slow infusion of 5% sodium taurocholate, after abdominal closure. Hyperlipidemic pancreatitis group (HLP group): high fat stomach perfusion agent (20% lard +10% cholesterol +20% Twain 80+2% +1% sodium cholate propylthiouracil + water) after two weeks, the pancreatic duct puncture injection of 5% sodium taurocholate, the abdomen was closed. Vera Pammy treatment group (CCB group): 10 minutes of hyperlipidemic pancreatitis model after intraperitoneal injection of Vera Pammy 1mg/kg. before operation through the jugular vein or heart blood test the triglyceride level (triglyceride, TG); 24 hours after the surgery, the statistical number of deaths, all rats were sacrificed, observe and record the amount of ascites; by enzyme-linked immunosorbent assay (ELISA) determination of serum thromboxane A2 (TXA2) and platelet activating factor (PAF) content; proper amount of pancreatic tissue by hematoxylin eosin staining (HE staining), the pathological change of pancreas were observed under microscope, and the pathological score of pancreatic tissue injury; proper amount of pancreatic tissue calcium level by flow cytometry technique in cells was detected by Western blotting analysis of NF; -魏Bp65铔嬬櫧鐨勮〃杈

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