Age-Dependent Effects of UCP2 Deficiency on Experimental Acute Pancreatitis in Mice

Reactive oxygen species (ROS) have been implicated in the pathogenesis of acute pancreatitis (AP) for many years but experimental evidence is still limited. Uncoupling protein 2 (UCP2)-deficient mice are an accepted model of age-related oxidative stress. Here, we have analysed how UCP2 deficiency affects the severity of experimental AP in young and older mice (3 and 12 months old, respectively) triggered by up to 7 injections of the secretagogue cerulein (50 μg/kg body weight) at hourly intervals. Disease severity was assessed at time points from 3 hours to 7 days based on pancreatic histopathology, serum levels of alpha-amylase, intrapancreatic trypsin activation and levels of myeloperoxidase (MPO) in lung and pancreatic tissue. Furthermore, in vitro studies with pancreatic acini were performed. At an age of 3 months, UCP2^-/- mice and wild-type (WT) C57BL/6 mice were virtually indistinguishable with respect to disease severity. In contrast, 12 months old UCP2^-/- mice developed a more severe pancreatic damage than WT mice at late time points after the induction of AP (24 h and 7 days, respectively), suggesting retarded regeneration. Furthermore, a higher peak level of alpha-amylase activity and gradually increased MPO levels in pancreatic and lung tissue were observed in UCP2^-/- mice. Interestingly, intrapancreatic trypsin activities (in vivo studies) and intraacinar trypsin and elastase activation in response to cerulein treatment (in vitro studies) were not enhanced but even diminished in the knockout strain. Finally, UCP2^-/- mice displayed a diminished ratio of reduced and oxidized glutathione in serum but no increased ROS levels in pancreatic acini. Together, our data indicate an aggravating effect of UCP2 deficiency on the severity of experimental AP in older but not in young mice. We suggest that increased severity of AP in 12 months old UCP2^-/- is caused by an imbalanced inflammatory response but is unrelated to acinar cell functions.     

The Effects of Probiotic Supplementation on Experimental Acute Pancreatitis: A Systematic Review and Meta-Analysis

Background

In February 2008, the results of the PRObiotics in PAncreatitis TRIAl (PROPATRIA) were published. This study investigated the use of probiotics in patients suffering from severe acute pancreatitis. No differences between the groups were found for any of the primary endpoints. However, mortality in the probiotics group was significantly higher than in the placebo group. This result was unexpected in light of the results of the animal studies referred to in the trial protocol. We used the methods of systematic review and meta-analysis to take a closer look at the relation between the animal studies on probiotics and pancreatitis and the PROPATRIA-trial, focussing on indications for harmful effects and efficacy.

Methods and results

Both PubMed and Embase were searched for original articles concerning the effects of probiotics in experimental acute pancreatitis, yielding thirteen studies that met the inclusion criteria. Data on mortality, bacterial translocation and histological damage to the pancreas were extracted, as well as study quality indicators. Meta-analysis of the four animal studies published before PROPATRIA showed that probiotic supplementation did not diminish mortality, reduced the overall histopathological score of the pancreas and reduced bacterial translocation to pancreas and mesenteric lymph nodes. Comparable results were found when all relevant studies published so far were taken into account.

Conclusions

A more thorough analysis of all relevant animal studies carried out before (and after) the publication of the study protocol of the PROPATRIA trial could not have predicted the harmful effects of probiotics found in the PROPATRIA-trial. Moreover, meta-analysis of the preclinical animal studies did show evidence for efficacy. It may be suggested, however, that the most appropriate animal experiments in relation to the design of the human trial have not yet been conducted, which compromises a fair comparison between the results of the animal studies and the PROPATRIA trial.     

Impact of Global Fxr Deficiency on Experimental Acute Pancreatitis and Genetic Variation in the FXR Locus in Human Acute Pancreatitis

Background

Infectious complications often occur in acute pancreatitis, related to impaired intestinal barrier function, with prolonged disease course and even mortality as a result. The bile salt nuclear receptor farnesoid X receptor (FXR), which is expressed in the ileum, liver and other organs including the pancreas, exhibits anti-inflammatory effects by inhibiting NF-κB activation and is implicated in maintaining intestinal barrier integrity and preventing bacterial overgrowth and translocation. Here we explore, with the aid of complementary animal and human experiments, the potential role of FXR in acute pancreatitis.

Methods

Experimental acute pancreatitis was induced using the CCK-analogue cerulein in wild-type and Fxr^-/- mice. Severity of acute pancreatitis was assessed using histology and a semi-quantitative scoring system. Ileal permeability was analyzed in vitro by Ussing chambers and an in vivo permeability assay. Gene expression of Fxr and Fxr target genes was studied by quantitative RT-PCR. Serum FGF19 levels were determined by ELISA in acute pancreatitis patients and healthy volunteers. A genetic association study in 387 acute pancreatitis patients and 853 controls was performed using 9 tagging single nucleotide polymorphisms (SNPs) covering the complete FXR gene and two additional functional SNPs.

Results

In wild-type mice with acute pancreatitis, ileal transepithelial resistance was reduced and ileal mRNA expression of Fxr target genes Fgf15, SHP, and IBABP was decreased. Nevertheless, Fxr^-/- mice did not exhibit a more severe acute pancreatitis than wild-type mice. In patients with acute pancreatitis, FGF19 levels were lower than in controls. However, there were no associations of FXR SNPs or haplotypes with susceptibility to acute pancreatitis, or its course, outcome or etiology.

Conclusion

We found no evidence for a major role of FXR in acute human or murine pancreatitis. The observed altered Fxr activity during the course of disease may be a secondary phenomenon.     

空肠营养对重症急性胰腺炎的疗效观察

目的:探讨早期空肠营养对重症急性胰腺炎(SAP)的疗效。方法:118名重症急性胰腺炎患者随机分为EN组和TPN组,比较两组SAP病人住院时间、费用、感染率、并发症及死亡率等。结果:TPN组住院时间长,费用高,感染率、并发症及病死率高,差异具有显著性。结论:早期空肠营养支持可明显改善SAP病情,提高治疗效果。     

空肠营养对重症急性胰腺炎的疗效观察

目的：探讨早期空肠营养对重症急性胰腺炎（SAP）的疗效。方法：118名重症急性胰腺炎患者随机分为EN组和TPN组．比较两组SAP病人住院时间、费用、感染率、并发症及死亡率等。结果：TPN组住院时间长,费用高,感染率、并发症及病死率高。差异具有显著性。结论：早期空肠营养支持可明显改善SAP病情,提高治疗效果。     

Effect of Octreotide on Enteric Motor Neurons in Experimental Acute Necrotizing Pancreatitis

Background/Aims

Amelioration of intestinal dysmotility and stasis during the early period of acute necrotizing pancreatitis (ANP) appears to be important to reduce the risks of secondary pancreatic infection. We aimed to characterize the association between the neuropathy of the enteric nervous system and gut dysfunction and to examine the effect of octreotide on motor innervation in the early stage of ANP.

Methodology/Principal Findings

The rats were randomly divided into eight groups: control+saline; control+octreotide; ANP+saline and ANP+octreotide (24 h, 48 h, 72 h). The spontaneous activity of ileal segments and the response to ACh, l-NNA were recorded. The alterations of myenteric neuronal nitric oxide synthase (nNOS), choline acetyltransferase (CHAT), PGP9.5 and somatostatin receptor 2 (SSTR₂) immunoreactive cells were evaluated by immunofluorescence and the protein expression of nNOS and CHAT were evaluated by western blot. We found the amplitude of spontaneous contractions at 48 h and the response to ACh at 24 h declined in the ANP+saline rats. A higher contractile response to both ACh and to l-NNA was observed in the ANP+octreotide group, compared with the ANP+saline rats at 24 h. A significant reduction in the nNOS and cholinergic neurons was observed in ANP+saline rats at the three time points. However, this reduction was greatly ameliorated in the presence of octreotide at 24 h and 48 h. The protein expression of CHAT neurons at 24 h and the nNOS neurons at 48 h in the ANP+octreotide rats was much higher than the ANP+saline rats.

Conclusion

The pathogenesis of ileus in the early stage of ANP may be related to the neuropathy of the enteric nervous system. Octreotide may reduce the severity of ileus by lessening the damage to enteric motor innervation.     

黄芩甙对重症急性胰腺炎保护作用的研究

目的:探讨黄芩甙对重症急性胰腺炎保护作用的机制.方法:45只雄性SD大鼠随机分为假手术组、模型组和黄芩甙干预组,每组15只.假手术组进腹后仅翻动胰腺和十二指肠后关腹,模型组采用3.5%牛黄胆酸纳逆行胰胆管注射造成重症急性胰腺炎大鼠模型,黄苓甙干预组在造模成功后给予黄芩甙治疗.观察各组大鼠术后胰腺的病理变化,术后3 h、6h、12h腹水及血清淀粉酶、脂肪酶的变化,测定胰腺组织MDA,SOD,TNF-α及IL-8的含量.结果:①在各时间点,模型组腹水量、血清淀粉酶、脂肪酶均较假手术组显著增加(p＜0.01);黄芩甙干预组腹水量、血清淀粉酶、脂肪酶均较模型组显著减少(p＜0.05),其中12 h组腹水量较6h组显著减少(p＜0.05);6h、12h血清淀粉酶、脂肪酶较3 h组显著降低(p＜0.05);②假手术组组织结构完整,无明显改变;模型组胰腺及周围有皂化斑的形成,胰腺水肿、出血,炎细胞浸润,胰腺组织结构破坏明显,有大片坏死区;黄芩甙治疗组和模型组比较显著改善;③在各时间点,模型组胰腺组织中MDA,TNF-α及IL-8含量较假手术组均显著升高,SOD显著降低(p＜0.05);黄芩甙干预组较模型组MDA,TNF-α及IL-8含量显著降低,SOD显著升高(p＜0.05).结论:黄芩甙可能通过降低氧自由基对组织的损害,抑制炎症因子的产生和释放起到保护胰腺组织的作用.     

Effects of Soluble Epoxide Hydrolase Deficiency on Acute Pancreatitis in Mice

Background

Acute pancreatitis (AP) is a frequent gastrointestinal disorder that causes significant morbidity, and its incidence has been progressively increasing. AP starts as a local inflammation in the pancreas that often leads to systemic inflammatory response and complications. Soluble epoxide hydrolase (sEH) is a cytosolic enzyme whose inhibition in murine models has beneficial effects in inflammatory diseases, but its significance in AP remains unexplored.

Methodology/Principal Findings

To investigate whether sEH may have a causal role in AP we utilized Ephx2 knockout (KO) mice to determine the effects of sEH deficiency on cerulein- and arginine-induced AP. sEH expression increased at the protein and messenger RNA levels, as well as enzymatic activity in the early phase of cerulein- and arginine-induced AP in mice. In addition, amylase and lipase levels were lower in cerulein-treated Ephx2 KO mice compared with controls. Moreover, pancreatic mRNA and serum concentrations of the inflammatory cytokines IL-1B and IL-6 were lower in cerulein-treated Ephx2 KO mice compared with controls. Further, Ephx2 KO mice exhibited decreased cerulein- and arginine-induced NF-κB inflammatory response, MAPKs activation and decreased cell death. Conclusions -These findings demonstrate a novel role for sEH in the progression of cerulein- and arginine-induced AP.     

Vildagliptin-Induced Acute Pancreatitis

ObjectiveTo describe the first reported case of acute pancreatitis in a patient receiving vildagliptin.MethodsWe present the clinical, biochemical, and radiographic findings of the study patient.ResultsA 61-year-old woman who presented with severe abdominal pain was found to have acute pancreatitis. This occurred 5 weeks after the commencement of vildagliptin, a dipeptidyl-peptidase 4 inhibitor, for the treatment of type 2 diabetes mellitus. The patient’s pancreatic enzymes were elevated (amylase, 1205 U/L; lipase, 8846 U/L), and abdominal computed tomography demonstrated diffuse pancreatic swelling, cyst formation, and necrosis in the body of the pancreas. In the absence of an identifiable cause for the patient’s pancreatitis, vildagliptin was considered a potential trigger. The patient recovered after vildagliptin therapy was ceased.ConclusionsAlthough incretin-based therapy effectively treats type 2 diabetes mellitus, emerging reports of acute pancreatitis in patients receiving sitagliptin and exenatide have prompted the US Food and Drug Administration to issue an alert on these drugs. This appears to be the first reported case of acute pancreatitis in a patient receiving vildagliptin, and it supports the possibility that acute pancreatitis may be a rare effect of incretin-based therapy.(Endocr Pract. 2011;17:e48-e50)     

参芪扶正注射液对大鼠重症急性胰腺炎的治疗作用

目的:研究参芪扶正注射液对重症急性胰腺炎(SAP)的治疗作用。方法:66只Wistar大鼠随机分为:SAP假手术组(SO组),SAP模型组(SAP组)和参芪扶正注射液治疗组(SQ组),胆胰管逆行注射50g.l~(-1)牛磺胆酸钠复制大鼠SAP模型。造模成功后2、4、8、12、24 h检测血清中TNF-α、IL-6和ALT水平,观察肝脏组织病理改变。结果:与SO组相比,SAP组及SQ组血清中TNF-α、IL-6和ALT水平明显升高(P<0.05)。与SAP组相比,从8 h开始SQ组血清中TNF-α、IL-6和ALT水平开始下降(P<0.05),两组的肝脏病理改变无统计学意义。结论:参芪扶正注射液可以降低SAP时血清中ALT、TNF-α和IL-6水平,但不能改善肝脏组织的病理损伤程度,对SAP的发生、发展可能有一定的预防作用。     

丹参和甘遂在重症急性胰腺炎治疗中的联合应用

目的:探讨丹参和甘遂联合应用对重症急性胰腺炎(SAP)的治疗价值。方法:我院收治的SAP患者62例随机分为常规治疗组和联合治疗组(常规治疗加丹参和甘遂联合治疗)两组。比较两组间腹痛缓解时间、肠鸣音恢复时间、体温恢复时间、血淀粉酶恢复正常时间;局部并发症;ARDS、休克、肾功能衰竭、胰性脑病等全身并发症;平均住院时间、ICU住院时间、中转手术数和死亡例数。结果:与常规治疗组比较,联合治疗组中腹痛缓解时间、肠鸣音恢复时间、局部和全身并发症发生率、平均住院时间、ICU住院时间、死亡率显著减少(P<0.05),但体温恢复时间、血淀粉酶恢复正常时间和中转手术数无明显变化(P>0.05)。结论:在常规治疗基础上,联合应用丹参和甘遂对SAP具有缓解病情,降低平均住院时间和死亡率的作用。     

生长抑素联合奥美拉唑治疗重症胰腺炎的疗效分析

目的：探讨重症胰腺炎患者采用生长抑素联合奥美拉唑治疗的临床效果。方法：选取2005 年10 月至2012 年10 月医院收治的老年重症胰腺炎患者98 例。将所有患者随机分为观察组和对照组,每组各49 例,对照组患者采取常规方法治疗重症胰腺炎,观察组患者在此基础上加用生长抑素联合奥美拉唑进行治疗。两组患者的疗程均为7 天,治疗结束后,对两组患者的临床疗效、各项恢复指标和并发症等情况进行对比分析。结果：观察组患者治疗的总有效率显著高于对照组（93.9%vs.71.4%,P〈0.05）;观察组患者的平均住院、肠道恢复、腹痛腹胀缓解、血淀粉酶、尿淀粉酶等各项指标恢复至正常时间及预后均显著优于对照组,差异具有统计学意义（P〈0.05）。结论：采用生长抑素联合奥美拉唑治疗重症胰腺炎,可以获得更高的疗效,患者症状改善明显,治疗时间短,并发症少,死亡率低,值得临床推广。