丙酮酸乙酯对急性坏死性胰腺炎大鼠肺损伤的保护作用

目的:观察丙酮酸乙酯(EP)对急性坏死性胰腺炎(ANP)肺损伤大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和肺组织高迁移率族蛋白B1(HMGB1)mRNA表达的影响,探讨丙酮酸乙酯治疗急性坏死性胰腺炎肺损伤的机制.方法:逆行性胰胆管注射50 g/L牛磺胆酸钠制作ANP模型.随机分成3组,对照组、ANP组和EP治疗组(40 mg/kg,每隔6h静脉注射一次).ELISA法检测血清TNF-α和IL-1β水平;半定量逆转录聚合酶链反应(RT-PCR)法检测肺组织HMGB1 mRNA表达,并观察血氧变化及肺组织的病理变化.结果:ANP组血清TNF-α和IL-1β水平在建模后6h达高峰,12h下降,在此两时点治疗组血清TNF-α和IL-1β水平明显低于ANP组(TNF-α:131.6±29.6 ng/L vs 196.3±16.3 ng/L,65.0±16.6 ng/L vs 90.2±20.1 ng/L,P<0.05;IL-1β:194.9±26.8 ng/L vs 223.0±34.8 ng/L,105.2±24.0 ng/L vs 130.4±23.0 ng/L,P<0.05).ANP组大鼠肺组织HMGB1 mRNA表达水平在ANP后12h明显升高,至24h仍维持在高水平.治疗组肺组织HMGB1 mRNA表达水平在各时间点均明显低于ANP组(0.68±0.11 vs 0.88±0.11,0.81±0.11 vs 1.04±0.10,1.08±0.08 vs 1.33±0.15,P<0.05),且同期肺损伤比ANP组轻.治疗组PaO_2均明显高于ANP组.结论:丙酮酸乙酯能显著抑制TNF-α、IL-1β和HMGB1等早晚期炎症因子,改善低氧血症,对ANP肺损伤有明显保护作用.     

丙酮酸乙酯对D-氨基半乳糖诱导的急性肝损伤大鼠的保护作用

目的观察丙酮酸乙酯（EP）对D-氨基半乳糖盐酸盐诱导的急性肝损伤模型大鼠的保护作用,并探讨其作用机制。方法制备D-氨基半乳糖盐酸盐诱导的急性肝损伤大鼠模型48只,并随机分成正常组、模型组、小剂量EP提前干预组、大剂量EP提前干预组、小剂量EP治疗组和大剂量EP治疗组。在造模后24h时取血,采用ELISA法检测TNF-α、IFN-γ、IL-10和IL—18水平;采用RT—PCR法检测肝组织HMGB1mRNA水平变化。结果与正常组比较,模型组大鼠血清TNF-α、IFN-γ,IL-10和IL-18水平均无明显变化（P〉0．05）,而肝组织HMGB1mRNA水平明显升高（P〈0．01）;与模型组比较,EP提前干预组和EP治疗组动物血清TNF-α、IFN-γ,IL-10和IL-18水平均无明显变化,而EP提前干预则显著降低了肝组织HMGB1 mRNA水平（P〈0．01）,其中大剂量组最为显著（P〈0．01）;病理学检查显示,EP提前干预组大鼠肝组织学得到明显改善,尤其是在大剂量组,而且比EP治疗组更加显著。结论EP能有效保护D-氨基半乳糖盐酸盐诱导的大鼠急性肝损伤,保护效果与药物的干预剂量成正相关,并且提前干预的效果更明显。     

丙酮酸乙酯对实验性结肠炎大鼠结肠组织HMGB1表达及细胞因子的影响

目的:观察丙酮酸乙酯(EP)对实验性结肠炎大鼠结肠组织HMGB1表达及炎症因子的影响.方法:SD大鼠随机分为空白对照组、模型组、EP治疗组,每组12只;模型组、治疗组大鼠用DSS溶液复制实验性结肠炎大鼠模型,观察各组实验大鼠DAI评分、大体形态学及组织病理学评分(HPS)改变,采用应用逆转录-聚合酶链反应(RT-PCR)和免疫印迹法(Westernblot)检测结肠组织HMGB1表达;酶联免疫吸附法检测细胞因子(TNF-α、IL-6)的含量.结果:DAI和HPS在DSS模型组中高于空白对照组(7.20±2.28vs0.45±0.16,13.60±0.72vs6.4±0.85,P<0.01),在空白对照组结肠组织中HMGB1低表达,但在模型组表达显著上调(P<0.01);模型组血清TNF-α、IL-6水平高于空白对照组(190.40±24.55vs43.65±8.79,238.75±26.58vs74.3±7.92,P<0.01);与DSS模型组相比,EP可以缓解大鼠体质量的减轻,减少腹泻及便血的发生,并且能够显著下调结肠组织DAI、HPS、HMGB1mRNA和蛋白的表达,下调血清TNF-α、IL-6水平(P<0.05).结论:HMGB1参与了实验性结肠炎大鼠的发病过程,应用EP治疗可有效抑制实验性结肠炎大鼠结肠组织HMGB1的表达,可以明显下调TNF-α、IL-6等促炎因子的水平,有助于减轻实验性结肠炎大鼠炎症反应.     

重症急性胰腺炎多脏器功能衰竭的治疗护理

重症急性胰腺炎(severe acute pancreatitis，SAP)是由多种因素诱发，多个脏器受累的疾病。发病急，进展快，病情凶险，并发症多，可发生休克和多脏器官功能衰竭(MSOF)，甚至死亡。现代医学的发展已使SAI，的死亡率下降到10％左右。我们1999-06／2004-06共收治急性胰腺炎88例，男53例，女35例；平均年龄42．7岁。现将其中资料齐全的18例急性重症胰腺炎治疗护理情况作一总结，分析报告下。     

清胰Ⅱ号对重症急性胰腺炎大鼠胰腺损伤的保护作用及机制探讨

目的 观察清胰Ⅱ号对重症急性胰腺炎(SAP)大鼠胰腺的保护作用,并探讨其机制。方法 将56只SD大鼠随机分为3组,A、B组各24只采用胆胰管逆行注射5%牛磺胆酸钠建立SAP模型,C组8只开腹后仅翻动肠管。A组造模清醒后予以清胰Ⅱ号灌胃1次/6 h,共4次。造模后6、12、24 h,采用ELISA法检测血清高迁移率族蛋白1(HMGB1),光镜观察胰腺组织并行病理评分。结果 B组各时点血清HMGB1较C组升高(P均<0.01),胰腺病理评分增加(P均<0.01)。A组与B组同时点相比,血清HMGB1下降(P均<0.05),胰腺病理评分降低(P均<0.05)。结论 中药制剂清胰Ⅱ号能够降低血清HMGB1水平,从而降低胰腺组织损伤。     

重症急性胰腺炎合并多脏器功能障碍综合征

随着重症急性胰腺炎(SAP)的治疗水平不断提高,治愈率正在逐步上升,目前已达到80%-90%,临床上需要进一步攻克的难点不再是胰腺坏死,而是早期出现多脏器功能障碍(MODS)的问题。目前普遍关注的暴发性急性胰腺炎(fulminantacutepancreatitis,FAP)的核心问题也是早期出现的MODS。进一步研究、认识SAP所致多脏器功能障碍和衰竭的发生、发展规律与防治对策,对于提高SAP的治疗水平,降低死亡率意义重大。     

高迁移率族蛋白B1与重症急性胰腺炎的关系的研究进展

高迁移率族蛋白B1(HMGB1)作为重要的晚期炎性介质,在重症急性胰腺炎(SAP)时表达增加,通过自身毒性作用、与其他细胞因子的相互影响、对炎性细胞活性的改变、对血管内皮细胞的促炎活性作用以及对肠道屏障的损伤等机制参与SAP的发生、发展.HMGB1可成为治疗SAP的研究靶点之一.     

晚期炎症介质高迁移率族蛋白1与重症急性胰腺炎

重症急性胰腺炎（SAP）死亡率高，预后差。SAP的发生与大量细胞因子级联反应引起的全身炎症反应综合征（SIRS）密切相关。SIRS是造成多器官功能不全综合征（MODS）的重要原因。近10余年临床研究显示，早期释放的炎症因子如TNF—α、IL-1等均在模型建立后迅速升高至峰值，并迅速下降，而此时炎症反应仍在继续，并且临床应用TNF—α、IL-1受体拮抗剂并未取得显效果，提示可能存在晚期炎症介质参与病理反应。高迁移率族蛋白1（highmobility group box 1，HMGB1）是一类广泛存在于真核细胞内的非组核蛋白，近年来有关HMGB1的报道逐年增加。本就此做一简要综述。[第一段]     

NF-κB、HMGB1在重症急性胰腺炎肠黏膜损伤中的时点表达及其意义

目的:探讨NF-κB、HMGB1在SAP大鼠肠黏膜损伤发生发展过程中的时点表达规律及其意义.方法:将70只SD大鼠随机分为A组(n=40)和S组(n=30),再分别按3、6、12、24、36h时点随机均等分成5个亚组.A组大鼠行逆行胰胆管匀速泵入5%牛磺胆酸钠建立SAP模型,S组大鼠开腹仅翻动十二指肠.两组大鼠均在建模后按时点开腹,门静脉取血检测AMY、DAO浓度,ELISA法和免疫组化法检测小肠黏膜NF-κB和HMGB1的表达.结果:(1)A组大鼠血DAO浓度随时点延迟逐渐增加;(2)肠黏膜NF-κB表达在3h最高,随时点延迟逐渐下降,24h、36h降至正常水平;(3)肠黏膜HMGB1表达6h开始明显升高,且随时点延迟逐渐增高,在24h最高,一直持续到36h仍然保持在较高水平.结论:(1)SAP大鼠早期即出现肠黏膜损伤;(2)SAP肠黏膜早期的损伤可能与小肠组织NF-κB的表达增加有关;(3)HMGB1作为晚期炎症介质可能介导了SAP肠黏膜损伤的发生发展;(4)HMGB1的调控可能受到了NF-κB的调节.     

重症急性胰腺炎并发心肌损伤的研究进展

重症急性胰腺炎（SAP）是一种病情凶险，并发症多的急腹症，常并发胰外脏器损害，包括：心血管失代偿、急性呼吸窘迫综合征（ARDS）、消化道出血、急性肾功能衰竭、胰性脑病，甚至多脏器功能障碍综合征（MODS）。其中心脏损害是最严重的并发症之一，表现为心律失常和心功能不全。据报道，心血管失代偿是SAP时导致病死率最高的并发症。近年来有关SAP／并发心肌损伤的研究逐渐增多，本据此作一综述。     

Delayed ethyl pyruvate therapy attenuates experimental severe acute pancreatitis via reduced serum high mobility group box 1 levels in rats

AIM： To investigate the effect of delayed ethyl pyruvate （EP） delivery on distant organ injury, survival time and serum high mobility group box 1 （HMGB1） levels in rats with experimental severe acute pancreatitis （SAP）.
METHODS： A SAP model was induced by retrograde injection of artificial bile into the pancreatic ducts of rats. Animals were divided randomly into three groups （n = 32 in each group）： sham group, SAP group and delayed EP treatment group. The rats in the delayed EP treatment group received EP （30 mg/kg） at 12 h, 18 h and 30 h after induction of SAP. Animals were sacrificed, and samples were obtained at 24 h and 48 h after induction of SAP. Serum HMGB1, aspartate arninotransferase （AST）, alanine arninotransferase （ALT）, blood urea nitrogen （BUN）, and creatinine （Cr） levels were measured. Lung wet-to-dry-weight （W/D） ratios and histological scores were calculated to evaluate lung injury. Additional experiments were performed between SAP and delayed EP treatment groups to study the influence of EP on survival times of SAP rats.
RESULTS： Delayed EP treatment significantly reduced serum HMGB1 levels, and protected against liver, renal and lung injury with reduced lung W/D ratios （8.22 ±0.42 vs 9.76 ± 0.45, P 〈 0.01）, pulmonary histological scores （7.1 ± 0.7 vs 8.4 ± 1.1, P 〈 0.01）, serum AST （667 ± 103 vs 1 368 ± 271, P 〈 0.01）, ALT （446 ± 91 vs 653 ± 98, P 〈 0.01） and Cr （1.2 ± 0.3 vs 1.8 ± 0.3, P 〈 0.01） levels. SAP rats had a median survival time of 44 h. Delayed EP treatment significantly prolonged median survival time to 72 h （P 〈 0.01）.
CONCLUSION： Delayed EP therapy protects against distant organ injury and prolongs survival time via reduced serum HMGBllevels in rats with experimental SAP. EP may potentially serve as an effective new therapeutic option against the inflammatory response and multiple organ dysfunction syndrome （MODS） in SAP patients.     

Delayed ethyl pyruvate therapy attenuates experimental severe acute pancreatitis via reduced serum high mobility group box 1 levels in rats

AIM: To investigate the effect of delayed ethyl pyruvate (EP) delivery on distant organ injury,survival time and serum high mobility group box 1 (HMGB1) levels in rats with experimental severe acute pancreatitis (SAP).METHODS: A SAP model was induced by retrograde injection of artificial bile into the pancreatic ducts of rats. Animals were divided randomly into three groups (n=32in each group): sham group, SAP group and delayed EP treatment group. The rats in the delayed 18 h and 30 h after induction of SAP. Animals were sacrificed, and samples were obtained at 24 h and 48 h after induction of SAR Serum HMGB1, aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine (Cr) levels were measured. Lung wet-to-dry-weight (W/D) ratios and histological scores were calculated to evaluate lung injury. Additional experiments were performed between SAP and delayed EP treatment groups to study the influence of EP on survival times of SAP rats.RESULTS: Delayed EP treatment significantly reduced serum HMGB1 levels, and protected against liver, renal and lung injury with reduced lung W/D ratios (8.22±0.42vs 9.76±0.45, P < 0.01), pulmonary histological scores (7.1±0.7 vs 8.4±1.1, P < 0.01), serum AST (667± 103 vs 1 368 ± 271, P < 0.01), ALT (446±91vs 653±98, P < 0.01) and Cr (1.2 ± 0.3 vs 1.8±0.3,P < 0.01) levels. SAP rats had a median survival time of 44 h. Delayed EP treatment significantly prolonged median survival time to 72 h (P < 0.01).CONCLUSION: Delayed EP therapy protects against distant organ injury and prolongs survival time via reduced serum HMGBllevels in rats with experimental SAP. EP may potentially serve as an effective new therapeutic option against the inflammatory response and multiple organ dysfunction syndrome (NODS) in SAP patients.     

High-mobility group box 1 protein and its role in severe acute pancreatitis

The high mobility group box 1(HMGB1),which belongs to the subfamily of HMG-1/-2,is a highly conserved single peptide chain consisting of 215 amino acid residues with a molecular weight of approximately 24894 Da.HMGB1 is a ubiquitous nuclear protein in mammals and plays a vital role in inflammatory diseases.Acute pancreatitis is one of the most common causes of acute abdominal pain with a poor prognosis.Acute pancreatitis is an acute inflammatory process of the pancreas(duration of less than six months),for which the severe form is called severe acute pancreatitis(SAP).More and more studies have shown that HMGB1 has a bidirectional effect in the pathogenesis of SAP.Extracellular HMGB1 can aggravate the pancreatic inflammatory process,whereas intracellular HMGB1 has a protective effect against pancreatitis.The mechanism of HMGB1 is multiple,mainly through the nuclear factor-κB pathway.Receptors for advanced glycation endproducts and toll-like receptors(TLR),especially TLR-2 and TLR-4,are two major types of receptors mediating the inflammatory process triggered by HMGB1 and may be also the main mediators in the pathogenesis of SAP.HMGB1 inhibitors,such as ethyl pyruvate,pyrrolidine dithiocarbamate and Scolopendra subspinipes mutilans,can decrease the level of extracellular HMGB1 and are the promising targets in the treatment of SAP.     

Effects of emodin and baicalein on rats with severe acute pancreatitis

AIM: To investigate the therapeutic effects of emodin in combination with baicalein on severe acute pancreatitis (SAP) rats and to explore the mechanism of SAP. METHODS: A total of 112 SAP rats induced by retrograde injection of 5% sodium taurocholate into the biliary-pancreatic duct, randomly assigned to a untreated group and three treated groups emodin group, combined emodin and baicalein group, and sandostatin group. Meanwhile, another 28 other rats were selected as sham operation (SO) group. There were 28 rats in each group, 8 rats were in 3 and 6 h groups respectively, and 12 rats in 12 h group. At each time-points, survival rates,ascites volumes, pathological lesion scores of pancreas tissues,serum amylase, tumor necrosis factor-α and IL-6 levels were determined as the indexes of therapeutic effects. RESULTS: The survival rate at 12 h was significantly higher in three treated groups than in untreated group.The ascites volume at 12 h was remarkably less in combined and sandostatin groups than in emodin group,but there was no difference between combined group and sandostatin group (P>0.05). Serum amylase levels at all time-points were significantly lower in three treated groups than in untreated group. However, they had no difference among treated groups (P>0.05).Serum TNF-α were lower in three treated groups than in untreated group at all time points. Among the three treated groups, at 6 h, the TNF-α levels of combination and sandostatin groups were lower than those of emodin group. These was no difference between combined and sandostantin. Serum IL-6 concentration at 3 h were lower in combined and sandostatin groups than in untreated group, but at 6 and 12 h they were lower in all treated groups than in untreated group and the combined and sandostatin groups and in emodin group, no difference was found between combined and sandostatin groups at all time-points (P>0.05). The pathological scores of pancreas at all time points were significantly lower in three treated groups than in the untreated group, and at 6, 12 h, the scores of combined and sandostatin groups were lower than in emodin group. There was no difference between combined and sandostatin groups (P>0.05). CONCLUSION: Combination of emodin with baicalein has significant therapeutic effects on SAP rats.     

Abdominal paracentesis drainage attenuates intestinal inflammation in rats with severe acute pancreatitis by inhibiting the HMGB1-mediated TLR4 signaling pathway

BACKGROUND Our previous studies confirmed that abdominal paracentesis drainage(APD)attenuates intestinal mucosal injury in rats with severe acute pancreatitis(SAP),and improves administration of enteral nutrition in patients with acute pancreatitis(AP).However,the underlying mechanisms of the beneficial effects of APD remain poorly understood.AIM To evaluate the effect of APD on intestinal inflammation and accompanying apoptosis induced by SAP in rats,and its potential mechanisms.METHODS SAP was induced in male adult Sprague-Dawley rats by 5%sodium taurocholate.Mild AP was induced by intraperitoneal injections of cerulein(20μg/kg body weight,six consecutive injections).Following SAP induction,a drainage tube connected to a vacuum ball was placed into the lower right abdomen of the rats to build APD.Morphological changes,serum inflammatory mediators,serum and ascites high mobility group box protein 1(HMGB1),intestinal barrier function indices,apoptosis and associated proteins,and toll-like receptor 4(TLR4)signaling molecules in intestinal tissue were assessed.RESULTS APD significantly alleviated intestinal mucosal injury induced by SAP,as demonstrated by decreased pathological scores,serum levels of D-lactate,diamine oxidase and endotoxin.APD reduced intestinal inflammation and accompanying apoptosis of mucosal cells,and normalized the expression of apoptosis-associated proteins in intestinal tissues.APD significantly suppressed activation of the intestinal TLR4 signaling pathway mediated by HMGB1,thus exerting protective effects against SAP-associated intestinal injury.CONCLUSION APD improved intestinal barrier function,intestinal inflammatory response and accompanying mucosal cell apoptosis in SAP rats.The beneficial effects are potentially due to inhibition of HMGB1-mediated TLR4 signaling.     

Abdominal paracentesis drainage ameliorates myocardial injury in severe experimental pancreatitis rats through suppressing oxidative stress

BACKGROUND Abdominal paracentesis drainage(APD) is a safe and effective strategy for severe acute pancreatitis(SAP) patients. However, the effects of APD treatment on SAPassociated cardiac injury remain unknown.AIM To investigate the protective effects of APD on SAP-associated cardiac injury and the underlying mechanisms.METHODS SAP was induced by 5% sodium taurocholate retrograde injection in SpragueDawley rats. APD was performed by inserting a drainage tube with a vacuum ball into the lower right abdomen of the rats immediately after SAP induction.Morphological staining, serum amylase and inflammatory mediators, serum and ascites high mobility group box(HMGB) 1, cardiac-related enzymes indexes and cardiac function, oxidative stress markers and apoptosis and associated proteins were assessed in the myocardium in SAP rats. Nicotinamide adenine dinucleotide phosphate oxidase activity and mRNA and protein expression were also examined.RESULTS APD treatment improved cardiac morphological changes, inhibited cardiac dysfunction, decreased cardiac enzymes and reduced cardiomyocyte apoptosis,proapoptotic Bax and cleaved caspase-3 protein levels. APD significantly decreased serum levels of HMGB1, inhibited nicotinamide adenine dinucleotide phosphate oxidase expression and ultimately alleviated cardiac oxidative injury.Furthermore, the activation of cardiac nicotinamide adenine dinucleotide phosphate oxidase by pancreatitis-associated ascitic fluid intraperitoneal injection was effectively inhibited by adding anti-HMGB1 neutralizing antibody in rats with mild acute pancreatitis.CONCLUSION APD treatment could exert cardioprotective effects on SAP-associated cardiac injury through suppressing HMGB1-mediated oxidative stress, which may be a novel mechanism behind the effectiveness of APD on SAP.     

Timing of mortality in severe acute pancreatitis： Experience from 643 patients

AIM： To determine the timing of mortality after onset of severe acute pancreatitis （SAP） and the course of the disease in a large series of patients. METHODS： From July 1996 to June 2005, all patients diagnosed with acute pancreatitis at Chang Gung Memorial Hospital, Taipei, Taiwan were retrospectively studied. Three thousand two hundred and fifty episodes of acute pancreatitis were recorded in 2248 patients （1431 males and 817 females; median age, 55.6 years; range, 18-97 years）. Mortality was divided into two groups： early death （≤ 14 d after admission）, and late death （〉 14 d after admission）. The clinical features of patients in these two groups were compared. RESULTS： Although the overall mortality rate of acute pancreatitis was 3.8% （123/3250）, mortality rate of SAP was as high as 16.3% （105/643）. Of those 105 SAP mortalities, 44 （41.9%） deaths occurred within the first 14 d after admission and 61 （58.1%） occurred after14 d. Incidence of early death did not significantly differ from that of late death. The co-morbidities did not contribute to the timing of death. Early deaths mainly resulted from multiple organ failure. Late deaths were mainly caused by secondary complication of infected necrosis. Intraabdominal bleeding significantly caused higher mortality in late death. CONCLUSION： Approximately half （42%） of SAP deaths occur within 14 d and most were due to multiple organ failure. The late deaths of SAP were mostly due to infected necrosis.