Pathological changes at early stage of multiple organ injury in a rat model of severe acute pancreatitis

BACKGROUND:Severe acute pancreatitis (SAP) is a commonly seen acute abdominal syndrome characterized by sudden onset,rapid progression and high mortality rate.The damage in peripheral organs may be more severe than that in the pancreas,and can even lead to multiple organ dysfunction.It is critical to recognize early pathological changes in multiple organs.This study aimed to assess the early pathological features of damaged organs in a rat model of SAP.METHODS:Thirty clean grade healthy male Sprague-Dawley ...     

急性胰腺炎大鼠肺组织中水通道蛋白-1的表达及功能

目的:研究水通道蛋白-1(AQP-1)在急性胰腺炎大鼠肺组织中的表达及其功能,探讨其表达与肺损伤的关系.方法:将Wistar大鼠分为假手术组(n=24)、肺损伤组(n=24)、地塞米松治疗组(n=24).采用逆行胰胆管注射15 g/L去氧胆酸诱发大鼠急性胰腺炎肺损伤模型,地塞米松组于造模后立即于尾静脉注射地塞米松2 mg/kg.每组分别于造模后4,8,12 h剖杀,取血及肺组织.通过检测血淀粉酶、血气、肺干/湿比值和肺组织病理切片判断胰腺炎及肺损伤的严重程度,放免法测血清TNF-α水平,RT-PCR检测肺组织AQP-1 mRNA的表达,免疫组化法检测肺组织AQP-1的表达.结果:与假手术组相比,胰腺炎肺损伤组血清淀粉酶、肺干/湿比值、TNF-α、肺组织病理损害程度明显升高,血氧、AQP-1mRNA(4 h:0.403±0.018 vs 0.794±0.015,P＜0.01;8 h:0.382±0.025 vs 0.812±0.032,P＜0.01;12 h:0.361±0.016 vs 198±5,P＜0.01)和AQP-1蛋白(4 h:104±4 vs 193±8,P＜0.01;8 h:96±5 vs 201±7,P＜0.01;12 h:94±3 vs198±5,P＜0.01)表达显著下调.与肺损伤组相比,地塞米松组血清淀粉酶、TNF-α、肺干/湿比值、肺组织病理损害程度明显降低,血氧、AQP-1 mRNA(4 h:0.681±0.031 vs 0.403±0.018,P＜0.05;8 h:0.763±0.013 vs 0.382±0.025,P＜0.05;12 h:0.784±0.032 vs 0.361±0.016,P＜0.05)和AQP-1的蛋白(4 h:145±6 vs104±4,P＜0.05;8 h:152±8 vs 96±5,P＜0.05;12 h:154±4 vs 94±3,P＜0.05)表达则明显升高,且与TNF-α的水平呈负相关性.结论:水通道蛋白-1表达与急性胰腺炎的肺损伤密切相关,其表达可能与TNF-α有关,地塞米松可上调其表达而减轻肺水肿.     

清胰汤对急性胰腺炎肺损伤治疗作用的实验研究

[目的]探讨清胰汤在重症急性胰腺炎（SAP）急性肺损伤（ALI）时对肺表面活性蛋白A（SP-A）表达及病情转归的影响。[方法]将SD大鼠随机分为3组，各10只。假手术（对照）组仅行剖腹术，模型组采用胆胰管内逆行注入1．5％去氧胆酸钠建立大鼠SAP时ALI模型，清胰汤治疗（治疗）组在建立SAP模型后30min、12h清胰汤（10ml／kg）灌胃。各组术后24h测动脉血pH、动脉氧分压（PaO2）、动脉二氧化碳分压（PaCO2）、血淀粉酶（AMY）、肺湿／干重（W／D）比值。应用RT-PCR检测肺sP-A mRNA的表达强度，并观察胰、肺病理变化。[结果]模型组AMY、W／D及PaCO2显著高于对照组和治疗组（均P〈0．01）。而模型组pH、PaO2显著低于其他2组（P〈0．05，〈0．01）。治疗组肺sP-A mRNA表达显著高于模型组（P〈0．01），其表达与肺损伤的程度呈负相关。治疗组胰、肺病理改变较模型组减轻。[结论]清胰汤能保护肺泡Ⅱ型上皮细胞功能，恢复SP-A mRNA正常表达，维持肺泡功能，从而对肺组织起保护作用。     

重症急性胰腺炎致急性肺损伤动物模型血气分析及形态学研究

目的观察sD大鼠重症急性胰腺炎致急性肺损伤后动脉血气分析及形态学改变。方法健康成年sD大鼠20只,随机分成两组,A组10只,注射同等剂量的生理盐水;B组10只,腹腔注射脂多糖建立重症急性胰腺炎的动物模型,观察病理学改变。结果血气分析变化：①PaO2：伤后12～48h两组sD大鼠相比较有差异（P〈0．05）,B组低于A组。②PaCO2：两组SD大鼠伤后12～48h相比较均有差异（P〈0．01）,B组高于A组。病理学改变：B组急性肺损伤表现（中性粒细胞浸润、肺泡内炎性物质渗出）,随着时间延长,视野内中性粒细胞增多。结论动脉血气指标PaO2、PaCO2在重症急性胰腺炎致急性肺损伤后可用作为判断肺组织损伤严重程度的参考指标,PaO2水平能反映肺损伤程度,在伤后24h～48h检测的指标更明显。     

Melatonin protects against pancreaticobiliary inflammation and associated remote organ injury in rats: role of neutrophils

Although the role of oxidative stress in acute pancreatitis (AP) has been studied in several animal models, little data are available regarding AP induced by pancreatic duct obstruction. We characterized the protective effects of melatonin on pancreaticobiliary inflammation and associated remote organ injury. In Sprague-Dawley rats, either the common pancreaticobiliary duct (PBDL; n = 28) or bile duct (BDL; n = 28) was ligated or a sham operation was applied (n = 14). Either melatonin (10 mg/kg) or vehicle (saline; 1 mL/kg) was administered intraperitoneally (i.p.) immediately before the surgery and twice a day until the rats were decapitated at 6 or 72 h. The pancreas, liver, kidneys and lungs were removed and tissue samples were stored for the determination of malondialdehyde (MDA) and glutathione (GSH) levels and myelopreoxidase activity. The results demonstrate that pathogenesis of acute obstructive pancreatitis involves not only the oxidative damage of the pancreatic and hepatic tissues, as assessed by increased MDA and reduced GSH levels, but the lungs and kidneys are also challenged by oxidant injury. Similarly, hepatic oxidative injury caused by cholestasis was also accompanied by pulmonary, renal and even pancreatic damage. The biochemical findings were also verified histologically. Melatonin, probably because of its free-radical scavenging and antioxidant activity, which involves an inhibitory effect on tissue neutrophil infiltration, protected all the affected tissues.     

MRI shows clodronate-liposomes attenuating liver injury in rats with severe acute pancreatitis

BACKGROUND:Studies have revealed that macrophages play an important role in the development of severe acute pancreatitis(SAP).Activated macrophages can lead to a systemic inflammatory response,induce lipid peroxidation,impair membrane structure,result in injury to the liver and the other extrahepatic organs,and eventually result in multiple organ dysfunction syndrome by promoting excessive secretion of cytokines.Liver injury can further aggravate the systemic inflammatory response and increase mortality by ...     

早期高容量血液滤过持续时间对重症急性胰腺炎急性肺损伤影响研究

目的 研究早期高容量血液滤过(HVHF)持续时间对重症急性胰腺炎(SAP)急性肺损伤(ALI)的影响.方法 将2006年8月到2009年4月怀化市第三人民医院ICU收治的49例入院时合并ALI急性呼吸窘迫综合征(ARDS)并在72 h内接受HVHF治疗的SAP患者随机分为两组.在常规治疗的基础上分别接受血滤持续时间8 h(Ⅰ组)和72 h(Ⅱ组)治疗.比较两组患者的APACHEⅡ评分、氧合指数、ALI/ARDS的改善率(包括治愈率)、机械通气的例数及时间、急性期并发症、HVHF相关并发症、结局及医疗费用等.结果 ①氧合指数及APACHEⅡ评分:两组入院第3天和第14天均较入院当天有所改善(P＜0.05).但在人院第3天和第14天,两组患者差异无统计学意义.②ALI、ARDS的改善率(包括治愈率):两组入院第3天和第14天较入院当天升高(P＜0.05);但在入院第3天和第14天.两组患者差异无统计学意义.③两组患者急性期机械通气的例数及时间、急性期并发症(多器官功能障碍综合征、急性肾功能衰竭、腹腔室隔综合征、导管相关感染、低血压)差异无统计学意义,但医疗费用差异有统计学意义(P＜0.05).两组患者急性期均无死亡.结论 发病72 h内的SAP早期短时(8 h)持续性HVHF治疗能有效促进合并ALI/ARDS的SAP患者肺功能的恢复,并且节约医疗费用.     

生大黄对重症急性胰腺炎并发肾损伤大鼠的治疗作用

[目的]探讨生大黄在大鼠重症急性胰腺炎(SAP)并发肾损伤时的治疗作用。[方法]将30只雄性SD大鼠随机分为假手术组、SAP组和治疗组,每组10只。采取4.5%牛磺胆酸钠胰胆管逆行注射制备大鼠SAP模型,治疗组在复制模型后即刻胃管内注入生大黄粉溶液。光镜下观察胰腺组织和肾组织的病理损害程度,测定肾脏组织湿干重比,检测各组大鼠血肌酐(Cr)和血尿素氮(BUN)含量,应用RT-PCR方法检测肾脏组织中肿瘤坏死因子-α(TNF-α)mRNA表达。[结果]治疗组大鼠肾脏湿干重比、Cr及BUN明显低于SAP组(P<0.01、<0.05、<0.05);假手术组未见TNF-αmRNA表达,与SAP组相比,治疗组TNF-αmRNA表达明显下降(P<0.05)。[结论]生大黄可以减轻胰腺组织和肾脏组织炎症程度,起到缓解病情、改善肾脏功能和促进修复的作用。     

Pulmonary function changes in rats with taurocholate‐induced pancreatitis are attenuated by pretreatment with melatonin

Melatonin is a free radical scavenger and broad‐spectrum antioxidant with immunomodulatory effects. We studied the effects of melatonin on changes in lung function, oxidative/nitrosative stress, and inflammatory cell sequestration in an acute pancreatitis (AP)‐associated lung inflammation model. Acute pancreatitis was induced by injection of 5% sodium taurocholate into the pancreatic duct of rats. Animals were randomized into control, AP, and a melatonin pretreatment (10 mg/kg)/AP group. Functional residual capacity (FRC), lung compliance (Cchord), expiratory flow rate at 50% (FEF50), airway resistance index (RI), and peak expiratory flow rate (PEF) were evaluated. White blood cell count (WBC) and hydrogen peroxide, lung lavage fluid WBC, methylguanidine, protein, lactic dehydrogenase (LDH), nitric oxide (NO), and leukotriene B4 (LTB4) levels were determined. Lung wet‐to‐dry weight ratio, peroxynitrite, and inducible nitric oxide synthase (NOS) mRNA and protein were measured. AP induction resulted in reductions in FRC, Cchord, FEF50, and PEF, and increase in RI and lung wet‐to‐dry weight ratio. Blood and lung lavage fluid WBC, lavage fluid LDH, protein, and blood hydrogen peroxide also increased. Levels of hydroxyl radicals, nitric oxide, and LTB4 in lung lavage fluid, inducible NOS mRNA, protein expression, and peroxynitrite in lung tissue also were significantly elevated. Pretreatment with melatonin attenuated obstructive and restrictive ventilatory insufficiency induced by AP. Blood and lavage WBC, lavage LDH and protein, lung edema, oxidative/nitrosative stress, and lipoxygenase pathway derivatives were also significantly attenuated by melatonin. We conclude that melatonin decreases AP‐induced obstructive and restrictive lung function changes via its antioxidant and anti‐inflammatory properties.     

Pathological changes in multiple organs of rats with severe acute pancreatitis treated by baicalin and octreotide

BACKGROUND:Severe acute pancreatitis(SAP)features fatal pathogenetic conditions and high mortality rate.The study of SAP complicated with multiple organ injuries is of important significance.In this study,we explored the protective effect of baicalin on multiple organs of SAP rats and compared it with that of octreotide through light and electron microscopic observations of the pathological changes. METHODS:The improved Aho method was used to prepare SAP rat models.These rats were then randomly divided into...     

Microcirculation disturbance affects rats with acute severe pancreatitis following lung injury

AIM: To study the effects of microcirculation disturbance (MD) on rats with acute severe pancreatitis (ASP). METHODS: We developed ASP rat models, and anatomized separately after 1, 3, 5, 7, and 9 h. We took out blood and did hemorrheologic examination and erythrocyte osmotic fragility test, checked up the water content, capillary permeability, and genetic expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissues, examined the apoptosis degree of blood vessel endothelium while we tested related gene expression of Bax and Bcl-2 in lung tissues. We did the same examination in control group. RESULTS: The viscosity of total blood and plasma, the hematocrit, and the erythrocyte osmotic fragility were all increased. Fibrinogen was decreased. The water content in lung tissues and capillary permeability were increased. Apoptosis degree of blood vessel endothelium was increased too. ICAM-1 genetic expression moved up after 1 h and reached its peak value after 9 h. CONCLUSION: MD plays an important role in ASP following acute lung injury (ALI). The functional damage of blood vessel endothelium, the apoptosis of capillary vessel endothelium, WBC edging-concentration and the increasing of erythrocyte fragility are the main reasons of ALI.     

大鼠急性胰腺炎相关肺损伤肺脏内源性H2S/CSE体系的变化

目的 观察急性胰腺炎相关肺损伤肺组织中内源性硫化氢(H2S)/胱硫醚γ裂解酶(CSE)体系的变化以及CSE抑制剂炔丙基甘氨酸(PAG)对急性胰腺炎相关肺损伤的影响.方法 54只SD大鼠被随机分为3组.①胰腺炎组:向胆总管中注射5％牛黄胆酸钠建立大鼠急性胰腺炎相关肺损伤模型;②药物干预组:在胰腺炎组的基础上,造模后lh腹...     

重症急性胰腺炎大鼠肺组织中LIF的表达变化及意义

目的:观察重症急性胰腺炎(SAP)大鼠白血病抑制因子(LIF)在肺组织中表达的时相变化, 探讨LIF在SAP病程及肺损伤中的意义．方法:36只♂SD大鼠随机分为正常对照组(N 组,n=6)、假手术组(Sham组,n=6)和重症急性胰腺炎组(SAP组,n=24)．采用胰管逆行灌注50 g/L牛磺胆酸钠的方法复制大鼠SAP模型．用RT-PCR法检测肺组织中LIF mRNA的表达水平,免疫组织化学方法检测NLIF在肺组织中的表达变化．结果:SAP组3 h后肺组织LIF mRNA的表达量明显高于对照组和假手术组(灰度值:1．018± 0．065 vs 1．451±0．067,1．322±0．072,P<0,05), 并且6,12,24 h持续升高(0．853±0．058,0．635 ±0．064,0．582±0．089)(P<0．01)．同样,SAP组 LIF蛋白表达在3和6 h后明显高于对照组和假手术(127．36±2．76,122．53±2．43 vs 159．46 ±2．78,156．35±3．12,P<0．05),并且12,24 h后也维持在很高的水平(109．37±2．87,102．42± 2．27)．结论:LIF作为促炎症因子参与了SAP肺组织的炎症反应．     

重症急性胰腺炎肺损伤内皮素的变化及丹参的治疗作用

目的:探讨重症急性胰膜炎(SAP)肺损伤时血清和肺组织内皮素(ET)变化及丹参的保护作用．方法:Wistar大鼠60只,随机分为假手术组(J 组)、模型组(F组)和丹参治疗组(D组)．50 g／L 牛磺胆酸钠胰胆管逆行注射方法制作SAP模型．D组大鼠分别在造模前1 d和造模后10 min ip丹参注射液(5 mL／kg)．各组在制模后24 h及 48 h测定血清ET-1水平及其在肺组织(光密度) 表达情况,同时观察肺系数变化及肺组织病理学改变．结果:与J组比较,F组24和48 h肺组织损伤明显加重,血清ET-1水平(F组:75．8±4．8,70．4± 4．8 ng／L;J组:32．0±6．9,30．3±4．8 ng／L)和肺系数显著增高(F组:0．62±0．06,0．73±0．07;J 组:0．41±0．08,0．41±0．07)(P<0．01),肺组织24 和48 h ET-1表达增高(F组:0．48±0．09,0.61± 0．10;J组:0．05±0．01,0．05±0．01)(P<0．01)．与 F组比较,D组24和48 h肺组织学损伤明显减轻,血清ET-1水平和肺系数明显下降(60．2± 7．3 ng／L,0．52±0．06,P<0．05;57．9±5．43 ng／L, 0．58±0．06,P<0．01),肺组织ET-1表达明显减少(0．23±0．10,0．36±0．09,P<0．01)．相关性分析显示,造模后24和48 h肺组织ET-1表达与肺系数密切相关(r=0．736,P<0．01;r=0．828, P<0．01)．结论:ET-1在SAP肺损伤中起着重要的作用, 丹参对SAP肺损伤具有保护作用．     

Effect of resveratrol on microcirculation disorder and lung injury following severe acute pancreatitis in rats

AIM: To investigate the mechanism of resveratrol underlying the microcirculation disorder and lung injury following severe acute pancreatitis (SAP). METHODS: Twenty-four rats were divided into 3 groups (SAP, sham and resveratrol groups) randomly. SAP model was established by injecting 4% sodium taurocholate l mL/kg through puncturing pancreatic ducts. Sham (control) group (8 rats) was established by turning over the duodenum. Resveratrol was given at 0.1 mg/kg b.m. intraperitoneally. Rats were sacrificed 9 h after SAP was induced. Blood samples were obtained for hemorrheological examination. Lung tissues were used for pathological observation, and examination of microvascular permeability, dry/wet ratio and myeloperoxidase (MPO) activity. Gene expression of intercellular adhesion molecule-1 (ICAM-1) was detected by RT-PCR. RESULTS: Compared with SAP group, resveratrol relieved the edema and infiltration of leukocytes in the lungs. Resveratrol improved markers of hemorrheology: high VTB (5.77±1.18 mPas vs9.49±1.34 mPas), low VTB (16.12±3.20 mPas vs30.91±7.28 mPas), PV (4.69±1.68 mPas vs 8.00±1.34 mPas), BSR (1.25±0.42 mm/h vs50.03±0.03 mm/h), VPC (54.67±3.08% vs 62.17±3.39%), fibrinogen (203.2?7.8 g/ L vs 51.3±19.1 g/L), original hemolysis (0.45±0.02 vs 0.49±0.02), and complete hemolysis (0.41±0.02 vs 0.43±0.02) (P<0.05). Resveratrol decreased the OD ratio of ICAM-1 gene (0.800±0.03 vs 1.188±0.10), dry/wet ratio (0.74±0.02 vs 0.77±0.03), microvascular permeability (0.079±0.006 vs 0.112±0.004) and MPO activity (4.42±0.32 vs 5.03±0.51) significantly (P<0.05). CONCLUSION: Resveratrol can improve the microcirculation disorder of the lung by decreasing leukocyte-endothelial interaction, reducing blood viscosity, improving the decrease of blood flow, and stabilizing erythrocytes in SAP rats. It may be a potential candidate to treat SAP and its severe complications (ALI).     

牛胆酸钠诱导大鼠急性胰腺炎系列模型的研究

目的 用牛胆酸钠纯化学制剂诱导大鼠实验性急性胰腺炎系列模型。 方法 用0.25％(n=4),1.0％(n=7),2.0％(n=7)或3.5％(n=7)牛胆酸钠溶液0.1 ml/100g逆行注入Wistar大鼠胰管内诱导急性胰腺炎。诱导急性胰腺炎前,及诱导后6,12 h分别取血分离血清,测定淀粉酶值。同时,观察各实验组胰腺病理组织光镜和电镜变化。 结果 随牛胆酸钠诱导浓度提高,血清淀粉酶逐渐升高,组织病理改变依次加重。1.0％和2.0％牛胆酸钠分别诱导轻度及重度水肿型胰腺炎;3.5％牛胆酸钠则诱导坏死型胰腺炎。 结论 胰腺病变程度与牛胆酸钠诱导浓度呈正相关,本系列模型适合评价药物的防治效应。     

Therapeutic effects of caspase-1 inhibitors on acute lung injury in experimental severe acute pancreatitis

AIM： To assess the therapeutic effect of Caspase-1 inhibitors （ICE-I） on acute lung injury （ALI） in experimental severe acute pancreatitis （SAP）.
METHODS： Forty-two SD rats were randomly divided into 3 groups： healthy controls （HC, n = 6）; SAP-S group （n = 18）; SAP-ICE-i group （n = 18）. SAP was induced by retrograde infusion of 5% sodium taurocholate into the bile-pancreatic duct. HC rats underwent the same surgical procedures and duct cannulation without sodium taurocholate infusion, in SAP-S group, rats received the first intraperitoneal injection of isotonic saline 2 h after induction of acute pancreatitis and a repeated injection after 12 h. In SAP-ICE-I group, the rats were firstly given ICE inhibitors intraperitoneally 2 h after induction of pancreatitis. As in SAP-S group, the injection was repeated at 12 h. Serum 1L-1β was measured by EUSA. Intrapulmonary expression of Caspase-1, IL-1β and IL-18 mRNA were detected by semi-quantitative RT-PCR. The wet/dry weight ratios and histopathological changes of the lungs were also evaluated.
RESULTS： Serum IL-1β levels in SAP-S group were 276.77 ± 44.92 pg/mL at 6 h, 308.99 ± 34.95 pg/mL at 12 h, and 311.60 ± 46.51 pg/mL at 18 h, which were increased significantly （P 〈 0.01, vs HC）. in SAP- ICE-I group, those values were decreased significantly （P 〈 0.01, vs SAP-S）. intrapulmonary expression of Caspase-1, IL-1β and IL-18 mRNA were observed in the HC group, while they were increased significantly in the SAP-S group （P 〈 0.01, vs HC）. The expression of IL-lβ and IL-18 mRNA were decreased significantly in the SAP- ICE-I group （P 〈 0.01, vs SAP-S）, whereas Caspase-1 mRNA expression had no significant difference （P 〉 0.05）. The wet/dry weight ratios of the lungs in the SAP-S group were increased significantly （P 〈 0.05 at 6 h, P 〈 0.01 at 12 h and 18 h, vs HC） and they were decreased significantly in the SAP-ICE-I group （P 〈 0.05, vs SAP-S）.Caspase-1 inhibitors ameliorated the severit     

Protective effect of clodronate-containing liposomes on intestinal mucosal injury in rats with severe acute pancreatitis

BACKGROUND: Severe acute pancreatitis (SAP) can result in intestinal mucosal injury. This study aimed to demonstrate the protective effect of clodronate-containing liposomes on intestinal mucosal injury in rats with SAP. METHODS: Liposomes containing clodronate or phosphate buffered saline (PBS) were prepared by the thin-film method SAP models were prepared by a uniform injection of sodium taurocholate (2 mL/kg body weight) into the subcapsular space of the pancreas. Sprague-Dawley rats were randomly divide...     

趋化因子CXCL11及其受体CXCR3在重症急性胰腺炎相关肺损害中的作用

目的:制作大鼠重症急性胰腺炎(severe acute pancreatitis,SAP)模型,检测不同时间点趋化因子CXCL11及其受体CXCR3在SAP肺组织中的动态变化,探讨他们在SAP肺功能损害过程中的作用.方法:48只SD大鼠,雌雄不限,随机分为2组:对照组(C组),SAP组(P组),每组24只.4%牛黄胆酸钠逆行胰胆管注射建立SAP大鼠模型,剂量为1mL/kg,C组打开腹腔后仅仅翻动胰腺组织数次.每组随机分为4个亚组,每个亚组6只.4个组分别在1、3、6、12h抽血、处死,留取组织标本.分别检测各不同时间点组的血清淀粉酶、肺湿干重比,胰腺组织、肺组织病理,免疫组织化学法检测肺CXCL11及CXCR3的表达,酶联免疫吸附试验(ELISA)检测血清中的CXCL11的水平.结果:P组各亚组血清淀粉酶值明显升高(P<0.01vsC组);肺湿干重比值:P组3、6、12h组较C组明显升高(P<0.05);胰腺组织、肺组织病理:3、6、12hP组肺组织损伤明显;免疫组织化学显示P组CXCL11与CXCR3蛋白表达较C组表达明显增强(P<0.05),ELISA显示:1、3、6、12hP组血清CXCL11蛋白较C组明显增高(P<0.01).结论:CXCL11/CXCR3可能参与大鼠SAP急性肺功能损害的发病过程.