急性胰腺炎时胰腺组织修复机制的研究

目的探讨急性胰腺炎后胰腺修复的机制。方法实验所用动物为NIH Swiss小鼠，分两组，分别是盐水对照组、急性胰腺炎组(AP组)。急性胰腺炎诱导采用蛙皮素腹腔注射，用常规病理评价胰腺的炎症程度；流式细胞仪检测细胞增殖状态；免疫组化方法检测胶原Ⅰ的表达，蛋白酶谱检测MMP-3的表达。结果病理检测结果：蛙皮素腹腔注射可诱导小鼠急性胰腺炎。在急性胰腺炎组小鼠AP诱导后8h胰腺组织损伤最严重，至第7天时组织损伤基本恢复正常。胰腺流式细胞仪检测发现AP诱导后细胞增殖状态分三个时期：早期的增殖活跃期、中期的增殖抑制期和后期的高增殖状态。胶原Ⅰ的检测显示：盐水对照组胶原Ⅰ有少量表达，AP时胶原Ⅰ表达增强。MMP-3的检测结果：盐水对照组基本无表达，在AP时酶的活性增高。结论急性胰腺炎后，胰腺组织可进行修复。在其修复过程中，既有组织细胞的增殖修复，又存在间质纤维化修复和塑性。     

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。     

中性粒细胞对大鼠急性重症胰腺炎胰腺腺泡凋亡的作用

目的探讨中性粒细胞在大鼠急性重症胰腺炎时对胰腺的病理损害及对胰腺腺泡凋亡的作用。方法氨甲蝶呤（MTX）腹腔注射制成大鼠低中性粒细胞模型，然后以脱氧胆酸胰胆管注射诱发急性重症胰腺炎，造模后0、3、6、9、24、48、72h随机分批处死动物，按各时间点检测胰腺组织髓过氧化物酶活性（MPO），血清淀粉酶，应用末端脱氧核苷酸转移酶介导的DUPT末端标记染色（TUNEL）和电镜技术检测胰腺腺泡凋亡情况，胰腺组织的病理学检查。结果实验组MPO在造模24h开始明显低于对照组（P〈0．01），胰腺组织的病理损害程度也较对照组轻。两组胰腺腺泡凋亡在造模3和6h均仅出现少量凋亡，在24和48h显著增多，以后逐渐下降；实验组胰腺腺泡凋亡指数明显高于对照组胰腺腺泡凋亡指数（P〈0．01）。结论适当地降低外周血的中性粒细胞水平能够增加胰腺腺泡凋亡，从而减轻急性重症胰腺炎时胰腺的病理损害程度。     

18F标记脱氧葡萄糖-正电子发射扫描成像在小鼠重症急性胰腺炎早期诊断和疗效评价中的应用

目的 评价18F标记脱氧葡萄糖-正电子发射扫描成像(18 F-FDG-PET)在小鼠重症急性胰腺炎早期诊断和疗效评价中的应用价值.方法 采用连续7次腹内注射雨蛙素(每次间隔1 h)及脂多糖制作小鼠重症急性胰腺炎模型,57只雌性ICR小鼠随机分为硼替佐米(PS-341)治疗组(注射脂多糖前0.5h腹内注射0.5 mg/kg PS-341),模型对照组[注射脂多糖前0.5h腹内注射50％二甲基亚砜(DMSO)],空白对照组(生理盐水制模).首次注射雨蛙素后8h将小鼠处死,每组3只行PET胰腺扫描,8只取胰腺组织测髓过氧化物酶(MPO),另外8只光学显微镜下观察小鼠胰腺的病理形态.结果 正常对照组PET扫描时胰腺不显影,与模型对照组比较,治疗组小鼠胰腺18F-FDG的吸收率(5.27±0.35,3.48±0.49)、胰腺MPO活性(3.46±0.28、1.82±0.54)均显著降低,两者之间差异有统计学意义(P＜0.05);胰腺组织的病理学也得到明显改善,进一步验证了18FFDG-PET在重症急性胰腺炎诊断和疗效评价中的应用价值.结论 18 F-FDG-PET能够动态监测重症急性胰腺炎的发展和转归,评价治疗效果.     

雨蛙素腹腔注射法建立小鼠急性坏死性胰腺炎模型的实验研究

目的　建立一种稳定的急性坏死性胰腺炎模型。方法　 36只雄性ICR小鼠用查随机数字表的方法随机分为对照组 (n =6 )和实验组 (n =30 )。实验组予以雨蛙素腹腔注射 ( 5 0 μg/kg) ,每小时注射 1次 ,共 7次 ,并分别于首次注射后 9h、18h、2 4h、4 8h和 72h各处死 6只 ;对照组予以腹腔注射等量的生理盐水 ,于注射后 18h处死。比较两组动物的血淀粉酶值、胰腺重量及胰腺组织学评分分值。结果　腹腔注射雨蛙素后 9h ,小鼠血淀粉酶值升高 ,胰腺重量增加 ,18h达峰值 ,2 4h后逐渐下降。二者与胰腺病理损害程度相一致。 18h胰腺间质炎性细胞浸润和腺泡细胞坏死明显 ,胰腺组织学评分值达到最大值 (P<0 .0 5 )。结论　大剂量雨蛙素腹腔注射能诱导ICR小鼠产生急性坏死性胰腺炎。该模型制备方法简单 ,无创伤性 ,模型稳定 ,重复性好     

胰管栓塞治疗急性坏死性胰腺炎

急性坏死性胰腺炎疗效仍不满意。作者在8年间收治794例急性胰腺炎,其中89例为坏死性,死亡率达52.8%。近年来采用胰管栓塞法治疗慢性胰腺炎获成功,能否用此方法治疗急性坏死性胰腺炎,作者进行了动物实验。选17支体重为10～15kg的杂种狗,将自体胆汁按0.5ml/kg注入胰管,诱发急性坏死性胰腺炎,其中5支为对照组。实验组12支狗经胰管注入栓塞剂氯丁橡胶60IA。然后定期做胰腺病理检查、测定血清淀粉酶、脂肪酶和胰蛋白酶抑制物。对照组动物于术后1.5～4天全部死亡。存活动物在胰管栓塞疗法后7天、2周、1、2、4、6个月进行胰腺活组织检查。实验组动物症状改善,进食较早,体重恢复。栓塞后7天再次剖腹,发现腹腔已无渗液,坏死灶有肉芽组织生成,成纤维细胞增生活     

大鼠急性胰腺炎肠道细菌易位的实验研究

目的本实验是观察大鼠急性胰腺炎后经胃肠道给予庆大霉素或谷氨酰胺对肠道细菌易位的影响。方法结扎封闭群 Wistar 大鼠的胆管,制成急性胰腺炎模型。分假手术组、急性胰腺炎组、庆大霉素组和谷氨酰胺组4组,各组又分5个亚组,分别于术后24、48、72、96和144小时处死动物。无菌下取回盲部淋巴结、胰腺、脾、肝、门静脉血和盲肠内容做细菌培养、计数。胰腺和小肠组织做病理检查。结果表明大鼠急性胰腺炎发生后,肠系膜淋巴结细菌数和阳性率明显增高,与门静脉血比较有显著差异。庆大霉索组盲肠内容和肠系膜淋巴结的大肠杆菌数明显减少,革兰氏阳性菌明显增多。谷氨酰胺组盲肠内容和肠系膜淋巴结的细菌数均显著减少。结论大鼠急性胰腺炎发生后早期肠道细菌易位主要经肠系膜淋巴途径。经胃肠道给予谷氨酰胺可防止肠道内细菌易位,减少胰腺感染的发生。     

大鼠急性胰腺炎血浆TNF—α及病理的动态变化

为探索急性胰腺炎（ＡＰ）１～１２小时内外周血ＴＮＦ－α的变化及其与组织学的关系，用ｃｈｅｔｔｙ法制成大鼠急性胰腺炎模型，术后１小时、３小时、６小时、１２小时测血浆ＴＮＦ－α，取胰组织作组织学观察。结果发现：胰腺炎组各时相ＴＮＦ－α较对照组均明显增高；胰腺病理改变随时间延长而加重；但动态观察提示血浆ＴＮＦ－α与胰腺病理改变并非呈平行关系。结果提示ＴＮＦ－α在ＡＰ病程演变中起着一定的作用。     

Kupffer细胞通过Toll样受体4信号通路调控组织因子及组织因子相关微粒表达诱导急性胰腺炎的发生

目的探讨Kupffer细胞(KCs)在受到脂多糖刺激时是否释放出组织因子膜微粒(TF-MP),判断Toll样受体4(TLR4)通路能否调控KCs表达组织因子(TF)诱导急性胰腺炎的发生。方法采用野生型C57/BL6小鼠(WT小鼠)和TLR4-/-小鼠建立急性胰腺炎模型后向腹腔内注射脂多糖10 mg/kg,分别于6、12、24 h处死两种小鼠各5只,观察胰腺组织病理损伤情况及其中TF表达情况,同时提取各小鼠KCs,检测KCs中的TF和TLR4的蛋白及m RNA表达量。剩余10只WT小鼠和10只TLR4~-/-小鼠用于观察生存情况。另外,提取WT小鼠和TLR4~-/-小鼠肝脏KCs,加入脂多糖300μg/L刺激,检测0、15、30、60、120 min时各时间点KCs中TF和TLR4的蛋白及m RNA表达量,同时测定其上清液中TF和TF-MP水平。结果 TLR4~-/-小鼠胰腺病理损伤程度较WT小鼠明显减轻,胰腺组织和肝脏组织中TF蛋白表达明显低于WT小鼠(P0.05)。建立急性胰腺炎模型后,TLR4-/-小鼠的生存率明显高于WT小鼠(P0.05)。无论是动物实验还是细胞实验,TLR4~-/-小鼠KCs中TLR4和TF m RNA及蛋白表达量均明显低于WT小鼠(P0.05);另外,TLR4~-/-小鼠KCs上清液中TF和TF-MP水平在30、60及120 min时均较WT小鼠明显降低(P0.05)。结论 KCs可能通过TLR4信号通路调控TF及TF-MP的表达而诱导急性胰腺炎的发生。     

吴茱萸次碱治疗重症急性胰腺炎的实验研究

目的：探讨吴茱萸次碱对大鼠重症急性胰腺炎（SAP）的疗效及其机制。 方法：50只SD大鼠随机均分为假手术组和4个实验组,实验组大鼠以5%牛磺胆酸钠胰胆管逆行注射制作SAP模型后,分别给予生理盐水、乌司他丁、吴茱萸次碱、乌司他丁+吴茱萸次碱处理。术后24 h,行病理学检查,并检测各组血清淀粉酶活性及血浆与胰腺组织和内皮素1（EF-1）、降钙素基因相关肽（CGRP）水平。 结果：除假手术组外,各实验组大鼠均胰腺组织均出现不同程度的胰腺炎病理改变;与假手术组比较,各实验组均出现不同程度的血清淀粉酶活性升高,血浆与胰腺组织ET-1浓度增加,而血浆与胰腺组织CGRP含量在吴茱萸次碱处理组与联合用药组均明显升高（均P<0.05）,其余实验组无明显改变（均P>0.05）;实验组中,各药物处理组胰腺病变程度、血清淀粉酶活性、血浆与胰腺组织ET-1浓度均较生理盐水处理的模型组明显降低,且均以联合用药组最为明显（均P<0.05）。 结论：吴茱萸次碱对大鼠SAP具有治疗作用,其作用可能与增加CGRP水平从而改善胰腺组织微循环有关。     

Extrapancreatic multiorgan injury in a severe sublethal acute pancreatitis model

In order to develop a new severe but sublethal acute pancreatitis model for the study of clinically relevant extrapancreatic multiorgan injury, we have induced acute pancreatitis in a rat model by intraductal injection with low dose and moderate concentration of bile acid under low pressure. We examined the structural and functional features in the pancreas, lung, liver and kidney. The animals were divided into two groups: the bile acid injection group and the control group. In the bile acid injection group, acute necrotizing pancreatitis was induced by intraductal administration of 0.2 ml of 2.0% bile acid under 30 cm H2O pressure, while the controls underwent the sham operation. The two groups were divided into six subgroups (8 rats for each) and sacrificed at 12, 24, 36, 48, 72 and 144 h, respectively. The pancreatitis induced hyperamylasemia, ascites, pancreatic oedema, haemorrhage, acinar cell necrosis and extensive fat necrosis without early mortality. Accompanied with the pancreatic injury, the function and histologic changes have developed continuously in the kidney and liver for 72 and 144 h in the bile acid injection animals respectively. No pancreatitis associated pulmonary changes were found. Taking into account the results with the two previously developed models of pancreatitis, we conclude that the extrapancreatic injury in acute pancreatitis is found in the liver, kidney and lung, in that order, depending on the severity of pancreatitis. The present sublethal pancreatitis model, in comparison with the two previously studied acute pancreatitis models, is perfect for pathogenetic and therapeutic study of liver and renal changes in acute necrotizing pancreatitis.     

环孢素A对大鼠急性出血坏死性胰腺炎脑损伤保护作用

目的探讨环孢素A在大鼠急性出血坏死性胰腺炎（ANP）时对脑组织损害的保护作用，从而为胰腺炎脑损伤的治疗提供实验依据。方法将60只SD大鼠随机分为正常对照组，诱导组和环孢素A处理组，经胰胆管逆行注射5％牛磺胆酸钠建立大鼠ANP模型，12h后处死动物，测定血清的TNFα水平，并检测脑组织含水量、脑组织MDA含量及脑微血管内白细胞聚集及附壁现象。结果经环孢素A处理后，血清中TNFα水平、脑组织含水量、脑组织MDA含量、脑微血管内白细胞聚集及附壁现象显著降低（P〈0．01）。结论环孢素A可以减轻脑损害的发生和发展。     

灌注联合胰胆管结扎诱导大鼠急性胰腺炎

目的 模拟急性胆源性胰腺炎的发病条件,制作一种符合临床特点的大鼠急性坏死性胰腺炎模型.方法 设计大鼠胰胆管单扎和双扎两种结扎方法,测压并用2%甘氨脱氧胆酸(GDCA)以35 cm H2O(1 cm H2O=0.098 kPa)压力恒压灌注5 min,8、16、24 h剖杀,观察急性胰腺炎的发生情况.结果 单扎组胆胰管压力为:(20.60±1.51)cm H2O,双扎组为:(29.37±0.87)cmH20.两组均诱导急性胰腺炎,于24 h表现出坏死性胰腺炎特征.病理学评分显示,随时间进程,胰腺炎逐渐加重(P＜0.01).在各时间点,胆胰管双扎组炎症重于单扎组(P＜0.01).结论 恒压灌注2%GDCA联合胰胆管单扎和双扎均可诱导出大鼠急性坏死性胰腺炎;胰管压力及其持续时间与急性胰腺炎发生、发展密切相关.

Abstract：

Objective To establish a new acute necrotizing pancreatitis model in rats. Methods We designed single pancreatic bile duct ligation and double pancreatic bile duct ligation in rats, combined with retrogradely infusing 2% glycodeoxycholic acid ( GDCA), to test duct pressure and observe the severity of acute pancreatitis at 8, 16 and 24 h, respectively. Results In two groups, the duct pressures were (20. 60 ± 1.51 ) cm H2O and (29. 37 ±0. 87) cm H2O, respectively. Both of the two methods could induce typical acute necrotizing pancreatitis at 24 h, the severity of which progressively developed with time course(P ＜0. 01 ), at each time point, the severity of double duct ligation group was higher than single duct ligation group ( P ＜ 0. 01 ). Conclusion Retrogradely infusing 2% GDCA with constant pressure,combined with the two methods of pancreatic bile duct ligation, coude induce clinically relevant acute necrotizing pancreatitis in rats; the pancreatic duct pressure and its duration might be closely related to the arising and developing of acute pancreatitis.     

胰管引流治疗犬出血坏死性胰腺炎的实验研究

采用逆行加压胰管注射犬自身胆汁建立急性出血坏死性胰腺炎（ＡＨＮＰ）模型，并应用经十二指肠切开，胰管内放置引流治疗。结果表明：本方法治疗ＡＨＮＰ可提高实验犬的生存率，降低死亡率保持胰管引流通畅可减轻实验大胰腺的病理变化，促进其修复。对临床治疗有指导意义。     

Role of pancreatic duct occlusion with prolamine (Ethibloc) in necrotizing pancreatitis

There is evidence that the pancreatic duct plays an important role in the evolution of necrotizing pancreatitis. We hypothesized that occlusion of the pancreatic duct and its smaller ductules with prolamine (Ethibloc) in opossums at risk of severe necrotizing pancreatitis would have a beneficial effect on the progression of the disease. Sixteen opossums underwent bile duct ligation below the entrance of the pancreatic duct. They were divided into four groups at 6 days. Group I (control, n = 5) opossums were killed for histologic observation of the degree of necrosis of the pancreas; group II (n = 5) underwent external drainage of the pancreatic biliary duct; group III (n = 4) had external biliary drainage and ligation of the pancreatic duct; group IV (n = 7) was treated with external biliary drainage and ligation of the main pancreatic duct after instillation of prolamine. Serum amylase, lipase, and calcium values were determined. The pancreas was examined by inspection and histologically at the time of death, and the severity of the disease was determined by quantitation of pancreatic tissue necrosis. All animals in groups II and III died 8 to 14 days after bile duct ligation, and all had severe necrotizing pancreatitis. All animals in group IV survived and were killed at 2 to 10 weeks after prolamine (Ethibloc) injection into the pancreatic duct. A mild edematous pancreatitis was observed in all seven animals. Prolamine (Ethibloc) provided dramatic protection from progressive necrosis. This study does not provide an explanation, but it allows for speculation that ductal injection interrupted the deleterious effect of proteolytic enzymes and their leakage into the interstitial space of the pancreas.     

HMGB1 A box蛋白对于急性胰腺炎脏器损伤的保护作用

目的 应用高迁移率族蛋白-1(HMGB1)的拮抗剂-A box蛋白治疗小鼠急性胰腺炎模型并观察其疗效.探讨A box蛋白对于急性胰腺炎所致脏器损伤影响的作用机制.方法 以20%L-精氨酸腹腔注射诱导建立小鼠急性坏死性胰腺炎模型,并于建模后应用A box蛋白进行治疗,测定血AST、ALT、LDH、BUN及Cr等生化指标.比较对照组和治疗组小鼠的胰腺、肺、肾、肝的病理变化及两组小鼠的存活率和存活时间.结果 建模后48 h,治疗组小鼠的胰腺、肺组织损伤病理评分较对照组减少,有显著性差异(P<0.05),治疗组肾、肝组织损伤的病变程度较对照组减轻.建模后24 h治疗组的AST值较对照组明显降低(P<0.05),建模后48 h治疗组的AST、ALT、LDH和Cr值较对照组明显降低(P<0.05).治疗组小鼠的存活率(66.7%)明显高于对照组的存活率(26.7%)(P<0.05).结论 A box蛋白对于胰腺炎及其所引发的肺、肾、肝等脏器的损伤有显著的保护作用.可提高重症胰腺炎小鼠的存活率.     

急性出血坏死性胰腺炎大鼠肠黏膜PIgR的表达变化

目的了解急性出血坏死性胰腺炎（acute hemorrhagic necrotizing pancreatitis,AHNP）时肠黏膜多聚免疫球蛋白受体（polymeric immunoglobulin receptor,pIgR）表达变化及意义。方法将60只Sprague-Dawley大鼠随机分为假手术组和AHNP组,每组各30只,采用胰胆管内逆行注射5%牛磺胆酸钠溶液诱导大鼠AHNP模型。假手术组行开腹手术翻动肠管后关腹。造模成功12h后处死大鼠,收集胰腺和小肠标本,进行病理检查,检测小肠内容物分泌型免疫球蛋白A（secretory immunoglobulin A,sIgA）含量,用Western blotting法检测小肠黏膜pIgR蛋白的表达。结果与假手术组比较,急性出血坏死性胰腺炎大鼠肠黏膜和胰腺发生病理损伤,小肠内容物sIgA含量显著下降（P〈0.05）,小肠黏膜pIgR的表达水平显著降低（P〈0.001）。sIgA与pIgR显著正相关（P〈0.01）。结论急性出血坏死性胰腺炎大鼠肠黏膜pIgR表达下调在sIgA所介导的肠黏膜免疫屏障功能障碍中可能起重要作用。     

Extrapancreatic organ manifestations in a moderately severe acute pancreatitis model

In order to study the extent of clinically relevant extrapancreatic organ injury in a moderately severe pancreatitis model, we examined the structural and functional features of the pancreas, lung, liver and kidney in a rat model simulating gallstone pancreatitis. The animals were divided into three groups: the low-ligation group, the high-ligation group and the control group, and sacrificed at 6, 24, 42, 60 and 96 h. In the low-ligation group, moderately severe acute pancreatitis was induced by the ligation of the common biliopancreatic duct plus intralipid intragastric injection, while controls underwent the ligation of the bile duct above the pancreas (the high-ligation group) or only sham operation (the control group) with fat injection. The pancreatitis induced hyperamylasemia, pancreatic oedema, haemorrhage, acinar cell necrosis and extensive fat necrosis. Accompanied with a peak value of serum amylase activity 24 h after the induction, the kidney changes developed, characterized by decrease in urine output, increase in serum urea and creatinine, and proximal convoluted tubular damage under electron microscope. There were no pancreatitis associated lung or liver changes. These results suggest that this model can be used to study the pathogenesis and therapy of renal injury during acute moderately severe pancreatitis.     

Does edematous pancreatitis exist in biliary-related pancreatitis? A hemodynamic and histologic study of the progression of bile-induced pancreatitis

The progression of pancreatitis induced in dogs by either single or hourly injections of two different bile solutions was monitored to determine whether acute necrotizing pancreatitis developed through an earlier mild interstitial form. In this model of biliary-related pancreatitis, acute interstitial pancreatitis could not be produced. The earliest lesion produced, although having the macroscopic appearance of edematous pancreatitis, was histologically a mild necrotizing form of the disease. If the bile solution was of sufficient concentration, then further injections resulted in progression of the pancreatitis from this mild form of scattered areas of focal acinar necrosis through coalescence of these areas to areas of parenchymal hemorrhage. Pancreatic blood flow, measured through its arterial inflow, increased during the earliest phase of the disease, but then decreased as the disease progressed.     

犬自身胆汁灌注胰管与急性胰腺炎病变程度的关系

为探讨自身胆汁灌注胰管与急性胰腺炎时胰腺病理改变的关系,采用３０只本地健康成年杂种犬,随机分为５组,每组各６只,以不同压力,不同持续时间灌注犬胰管,观察胰腺的组织学变化。