重症急性胰腺炎大鼠肠黏膜免疫屏障的变化及甘露糖的干预效果

目的探讨重症急性胰腺炎大鼠肠黏膜免疫屏障的变化情况及甘露糖的干预效果。方法将30只大鼠随机分为假手术组、胰腺炎组(重症急性胰腺炎组)和甘露糖组(重症急性胰腺炎+甘露糖干预组),建模后12 h,胰腺和肠黏膜组织进行HE染色观察病理学变化,测定血内毒素、淀粉酶、瘦素水平和分泌性免疫球蛋白(sIg)A、甘露糖结合凝集素(MBL)、白细胞介素(IL)-4水平。结果胰腺炎组和甘露糖组胰腺病理学评分和肠黏膜病理学评分均高于假手术组(P0.05),甘露糖组大鼠胰腺病理学评分和肠黏膜病理学评分低于胰腺炎组(P0.05)。胰腺炎组和甘露糖组血清内毒素、淀粉酶、瘦素水平均高于假手术组(P0.05),甘露糖组大鼠血清内毒素、淀粉酶、瘦素水平低于胰腺炎组(P0.05)。胰腺炎组和甘露糖组血清sIgA、IL-4水平均低于假手术组(P0.05),甘露糖组血清sIgA、IL-4水平高于胰腺炎组;胰腺炎组和甘露糖组血清MBL水平均高于假手术组(P0.05),甘露糖组血清MBL水平高于胰腺炎组(P0.05)。结论重症急性胰腺炎破坏肠黏膜免疫屏障,甘露糖具有保护重症急性胰腺炎大鼠肠黏膜屏障的作用,其机制可能为甘露糖能够增加血清sIgA、MBL、IL-4水平。     

复合益生菌制剂对急性坏死性胰腺炎大鼠的保护作用

背景:肠道细菌易位是重症急性胰腺炎(SAP)时胰腺坏死感染的主要来源,因此保护肠黏膜屏障对SAP的治疗具有重要意义。目的:探讨复合益生菌制剂对急性坏死性胰腺炎(ANP)大鼠肠黏膜屏障和胰腺损伤的保护作用。方法:50只SPF级大鼠随机分为假手术组(n=10)、ANP模型组(n=20)和益生菌干预组(n=20)。采用胰腺被膜下均匀注射牛磺胆酸钠制备ANP模型,干预组术前30 min以双歧杆菌四联活菌片溶液灌胃。术后6 h采集标本,检测血淀粉酶、二胺氧化酶(DAO)、TNF-α水平,观察胰腺组织病理学表现和末端回肠组织超微结构。结果:ANP模型组血淀粉酶、DAO、TNF-α水平和胰腺组织学评分均显著高于假手术组(P<0.05),益生菌干预组各项指标均较ANP模型组有所改善(P<0.05)。假手术组末端回肠黏膜结构完整;ANP模型组回肠黏膜上皮细胞微绒毛萎缩、排列稀疏,细胞间连接松弛;益生菌干预组微绒毛稍稀疏,细胞间连接紧密度较ANP模型组增高。结论:复合益生菌制剂对ANP大鼠具有保护作用,不仅能减轻肠黏膜损伤,保护肠黏膜屏障功能,还能减轻胰腺局部损伤和全身性炎症反应。     

高迁移率族蛋白B1在重症急性胰腺炎肠屏障损害中的作用

目的:探讨高迁移率族蛋白B1在重症急性胰腺炎(SAP)肠屏障损害中的作用及其机制.方法:采用逆行胆胰管注射50 g/L牛磺胆酸钠制备SAP大鼠模型.将56只Wistar大鼠随机分为正常对照组(Control组),SAP 3h,6h,12h,24h,48h 5个亚组,二硫代氨基甲酸吡咯烷处理组(PDTC组),每组8只.PDTC组于建模后24h取材.测定血浆内毒素(LPs)、二胺氧化酶(DAO)水平,用逆转录-聚合酶链反应(RT-PCR)方法检测肠组织高迁移率族蛋白B1(HMGB1)mRNA表达,用Western blot法检测肠组织HMGB1水平.结果:与对照组比较,SAP 6h组大鼠肠组织HMGB 1 mRNA表达显著增高(0.41±0.06 vs 0.26±0.03,P<0.01),12h呈现进一步升高趋势,24h达峰值(0.62±0.06),并持续至48h.PDTC干预可显著降低肠组织HMGB1 mRNA表达(0.35±0.06 vs 0.62±0.06,P<0.01).PDTC组较SAP 24h组肠组织HMGB1 mRNA表达,血浆LPS和DAO显著下降(HMGB1 mRNA表达:0.35±0.06 vs 0.62±0.06,P<0.01;LPS:0.433±0.120 KEU/L vs 0.852±0.232 KEU/L,P<0.01;DAO:0.65±0.12 kU/L vs 1.36±0.22 kU/L,P<0.01).结论:SAP时,肠组织内HMGB1表达上调;PDTC可以明显抑制SAP时肠组织内HMGB1表达.     

伴高脂血症性重症急性胰腺炎大鼠肠道细菌移位的实验研究

目的建立高脂血症重症急性胰腺炎(SAP)大鼠模型,观察比较伴或不伴高脂血症SAP大鼠的内毒素水平及细菌移位情况,以了解肠源性感染在伴有高脂血症SAP中的地位。方法40只雄性SD大鼠随机等分为4组:正常组、高脂血症组、重症急性胰腺炎组、高脂血症重症急性胰腺炎组。建立大鼠高脂血症及大鼠SAP模型,观察胰腺组织病理改变;检测大鼠血清淀粉酶及三酰甘油、门静脉血内毒素水平;取标本(门静脉血、腹水、肠系膜淋巴结及回盲末端肠管)进行细菌培养,鉴定其菌种并计算细菌移位率。结果SAP组及HSAP组胰腺组织病理见明显的水肿、炎性细胞浸润、出血、坏死等改变,评分值较正常组有显著升高(P0.05),其中HSAP组胰腺组织坏死程度较SAP组严重,评分值差异有统计学意义(P0.05)。HL组血清TG值较正常组明显升高(P0.05),HSAP组血清TG值较SAP组升高(P0.05);SAP组血清AMS值、血内毒素水平及细菌移位率较正常组显著升高(P0.05),HSAP组血清AMS值较SAP组明显降低(P0.05),HSAP组血内毒素水平及细菌移位率较SAP组升高(P0.05);SAP组及HSAP组各标本移位细菌种类与肠管相同。结论伴或不伴高脂血症SAP大鼠在发病早期均可发生肠道黏膜损伤、内毒素血症和细菌移位,伴有高脂血症SAP组较单纯SAP组更易发生细菌移位及内毒素血症,肠源性感染有可能是其病情加重的原因之一。     

重症急性胰腺炎NF-κB活化及维生素C干预治疗的实验研究

目的观察重症急性胰腺炎胰腺组织中NF-kB活化情况及VitC干预治疗对NF-kB活性的影响。方法54只雌性大鼠随机分为假手术组、重症急性胰腺炎组(SAP)、VitC预处理组(VitC SAP)。SAP模型经胆胰管内加压注射3.5%牛黄胆酸钠0.1ml／100g。VitC预处理组在建立模型前1h按VitC注射液1g／kg肌注给药。分别于建模后3h、6h、12h将大鼠分批处死,检测血淀粉酶,免疫组化法检测各组胰腺组织NF-kB的表达。常规病理并按Jan'S标准进行评分。结果VitC预处理组血清淀粉酶水平较SAP组在各个时间点均明显下降(3h,P<0.01,6h、12h,P<0.05)。VitC预处理组胰腺炎症范围、腺泡坏死程度及血管变化均较SAP组明显减轻,胰腺组织病理评分在各时间点明显低于SAP组(P<0.01或P<0.05)。假手术组可见胰腺组织NF-kB少量表达,VitC预处理组胰腺细胞NF-kB阳性率在各时间点均较SAP组明显减少(P<0.01或P<0.05)。结论抗氧化剂VitC能抑制胰腺细胞NF-kB活化,减轻SAP胰腺组织损害和降低血清淀粉酶水平,对SAP具有一定治疗作用。     

钙超载在大鼠血脂异常胰腺炎中的研究

[目的]探讨钙超载在血脂异常胰腺炎(HLP)发病机制中的作用,从而为维拉帕米治疗HLP提供理论依据。[方法]将80只SD大鼠随机分成4组,每组20只:即假手术(SO)组,重症急性胰腺炎(SAP)组,HLP组,维拉帕米(CCB)治疗组。24h后统计各组死亡率,随即处死大鼠,检测血三酰甘油(TG);采用ELISA检测胰腺组织三磷酸肌醇(IP3);采用流式细胞仪技术测定胰腺细胞内钙离子(Ca2+)水平;采用Western blot检测胰腺组织核转因子(NF)-κB p65蛋白表达;采用ELISA法检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6;苏木精-伊红染色对大鼠胰腺组织学损伤进行评估。[结果]1HLP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(P0.05);2SO组死亡率明显低于其他各组(P0.05),HLP组与SAP组、CCB组差异有统计学意义(P0.05);3SO组与SAP组、SAP组与HLP组、HLP组与CCB组胰腺组织内IP3比较均差异有统计学意义(P0.05);4 SAP组胰腺腺泡细胞Ca2+水平显著高于SO组(P0.05),HLP组Ca2+高于SAP组、CCB组(P0.05);5SAP组胰腺组织NF-κBp65表达较SO组显著升高(P0.05),HLP组较SAP组、CCB组显著升高(P0.05);6SO组血清TNF-α、IL-6浓度与SAP组相比差异有统计学意义(P0.05),HLP组较SAP组、CCB组明显升高(P0.05);7SAP组胰腺组织学评分较SO组,HLP组较SAP组、CCB组均明显升高(P0.05)。[结论]钙超载在HLP的发生发展中起着关键性作用,维拉帕米治疗HLP效果显著。     

连翘对重症急性胰腺炎大鼠肝组织中NF-κB和Foxp3表达的影响

目的探讨核因子NF-κB和Foxp3在重症急性胰腺炎（SAP）肝损伤中的作用及连翘对其表达活性的影响。方法雄性Wistar大鼠80只,随机分成假手术组（SO组）、SAP组和干预组,其中干预组分连翘高、中、低剂量组和阳性对照组（PDTC）。牛磺胆酸钠溶液在胰胆管远端注射造模,SO和SAP组于术后3、6、12 h,干预组于术后12 h处死大鼠,分别留取标本。测各组血淀粉酶（AMY）、ALT及TNFα水平,鲎试剂法测血浆内毒素水平,流式细胞术测外周血Treg百分数,对胰腺及肝脏进行病理学检查及评分,RT-PCR法检测肝脏组织中NF-κBmRNA和Foxp3mRNA表达量。组间比较采用单因素方差分析,进一步进行多重比较,采用LSD法进行统计学处理,各指标间相关性分析采用直线相关分析。结果与SO组比较,SAP组中各项指标均随时间升高,于12 h达高峰。与SAP12 h组相比,干预组（大鼠死亡率为0）肝脏组织中的NF-κBmRNA和Foxp3mRNA表达明显降低（P〈0.01）,与Treg呈正相关（r=0.738,P〈0.01）。随连翘剂量增加,AMY、ALT及TNFα水平均明显降低,肝脏和胰腺组织炎症明显减轻,高剂量组和阳性对照组相比较无明显差异（P〉0.05）。结论 NF-κB的激活参与SAP肝损伤的发生,连翘能显著降低NF-κB的活性及肝脏组织中NF-κBmRNA和Foxp3mRNA的表达,减轻SAP肝损伤的严重程度。     

血管活性肠肽对重症急性胰腺炎肠屏障保护作用的实验研究

目的 探讨血管活性肠肽(VIP)对重症急性胰腺炎(SAP)大鼠肠屏障的保护作用及其机制.方法 54只SD大鼠随机分成3组:假手术组(SO)、SAP组和VIP干预组,采用微泵逆行胰胆管注射4%牛磺胆酸钠制备SAP动物模型,SAP制模后5 min腹腔注射VIP 5×10-9nmol作为VIP干预组.各组在制模后1 h、6 h、12 h检测血浆内毒素水平,逆转录(RT)-PCR法及免疫组化法检测肠黏膜Toll样受体4(TLR4)表达,并对肠黏膜组织行电镜检查.结果 与SO组相比,SAP组血浆内毒素和肠黏膜TLR4表达在制模1 h即开始增高,并随制模时间的延长而进行性增高.两者具有明显的相关性.电镜检查肠黏膜有明显的病理损伤.VIP干预组与SAP组同时点相比,血浆内毒素和肠黏膜TLR4表达降低,病理损伤减轻,制模后6 h尤为明显.SO组肠黏膜未见TLR4表达.结论 VIP能有效保护SAP肠屏障功能,其机制可能与下调肠黏膜TLR4的表达有关.     

脂质过氧化损伤对高脂血症大鼠重症急性胰腺炎的作用及机制

目的:观察高脂血症对大鼠急性胰腺炎病情的影响,探讨脂质过氧化损伤在伴高脂血症重症急性胰腺炎(SAP)中的作用及其机制.方法:SD大鼠脂肪乳剂灌胃2 wk建立高脂血症模型,逆行胰胆管注射3.5%牛磺胆酸钠诱发SAP模型.将大鼠50只随机分为4组:正常组(n=10);高脂血症对照组(HL组,n=10);SAP组(n=15);伴高脂血症SAP(HAP,n=15).检测血清淀粉酶(AMS)、甘油三酯(TG)及胆固醇(CH)水平,并检测血清及胰腺组织的丙二醛(MDA)、超氧化物歧化酶(SOD)、黄嘌呤氧化酶(XOD)、一氧化氮(NO),观察胰腺组织病理改变.结果:HAP组胰腺组织病理改变较SAP组严重:HAP组血清及胰腺组织MDA、XOD水平显著高于SAP组(血清:30.76±2.67 nmol/mLvs 23.14±3.42 nmol/mL,55.72±10.49 U/L vs 45.78±8.98 U/L,均P<0.01:胰腺组织:4.33±0.48 nmol/mgprot vs 2.87±0.45 nmol/mgprot,5.57±0.63 U/gprot vs 4.33±0.79 U/gprot,均P<0.01):其血清及胰腺组织SOD、NO水平显著低于SAP组(血清:85.46±13.56 U/mLvs 97.16±13.77 U/mL,31.72±10.50μmol/Lvs 52.97±6.01μmol/L,均P<0.05;胰腺组织:22.65±3.85 U/mgprot vs 27.88±4.43 U/mgprot,均P<0.01;1.09±0.21 μmol/gprot vs 1.48±0.40μmol/gprot,均P<0.05).结论:高脂血症可加SAP的胰腺病理改变;脂质过氧化损伤可能在高脂血症加重SAP的机制中发挥重要作用.     

大鼠重症急性胰腺炎发病机制中p38丝裂原活化蛋白激酶的作用研究

目的探讨p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)特异性抑制剂SB203580对大鼠重症急性胰腺炎(SAP)的保护作用。方法以胆胰管逆行注射5%牛磺胆酸钠建立SD大鼠SAP模型。将45只SD大鼠随机分为3组,假手术组(SO组,n=15);SAP组(SAP组,n=15);SB203580治疗组(SB组,n=15)。术后3、6、12 h处死大鼠并取血。对胰腺进行病理组织学评分,测定大鼠腹水量。检测血清淀粉酶,ELISA法测定血清IL-6和IL-10水平。并采用免疫组化法观察大鼠胰腺组织p38 MAPK下游靶点P-ATF2表达情况。结果 SO组胰腺组织存在P-ATF2弱表达,SAP组各时间点P-ATF2表达水平明显升高(P<0.01),SB组各时间点表达水平较SAP组下降明显(P<0.01)。SAP组6、12 h时间点IL-6水平,3、6 h时间点IL-10水平,血清淀粉酶,腹水量明显高于SB组(P<0.05)。SAP组各时间点胰腺组织病理学积分显著高于SB组,差异有统计学意义(P<0.05)。结论阻断p38 MAPK信号传导通路可以明显缓解大鼠重症急性胰腺炎的病情。预示此信号传导通路可以为SAP的治疗提供一个有价值的标靶。     

Influence of dexamethasone on inflammatory mediators and NF-κB expression in multiple organs of rats with severe acute pancreatitis

AIM： To observe the therapeutic effects of dexamethasone on rats with severe acute pancreatitis （SAP） and investigate the influences of dexamethasone on the inflammatory mediators and NF-κB expression in multiple organs of SAP rats as well as the mechanisms involved.
METHODS： Ninety Sprague-Dawley （SD） rats with SAP were randomly divided into the model group （n = 45） and dexamethasone treatment group （n = 45）, and another 45 rats were selected for the sham operation group. All groups were randomly subdivided into the 3 h, 6 h and 12 h groups, each group containing 15 rats. The survival of all groups and pathological changes of multiple organs （liver, kidney and lung） were observed at different time points after the operation. The pathological score of multiple organs was carried out, followed by the determination of amylase, endotoxin and TNF-α contents in blood. The tissue microarray was used to detect the expression levels of NF-κB p65 protein in multiple organs. RESULTS： There was no marked difference between the model group and treatment group in the survival rate. The amylase content,of the treatment group was significantly lower compared to the model group at 12 h （P 〈 0.01, 7791.00 vs 9195.00）. Moreover, the endotoxin and TNF-α levels of the treatment group were significantly lower than that of the model group at 6 h and 12 h （P 〈 0.01, 0.040 vs 0.055, 0.042 vs 0.059 and P 〈 0.05, 58.30 vs 77.54, 38.70 vs 67.30, respectively）. Regarding the changes in liver NF-κB expression, the model group significantly exceeded the sham operation group at 3 h （P 〈 0.01, 1.00 vs 0.00）, and the treatment group significantly exceeded the sham operation group at 12 h （P 〈 0.01, 1.00 vs 0.00）, whereas no marked difference was observed between the model group and treatment group at all time points. The kidney NF-κB expression level in the treatment group significantly exceeded the model group （P 〈 0.05, 2.00 vs 0.00） and the sham operation group （P 〈 0.01, 2     

益生元对急性胰腺炎大鼠肠屏障功能的影响

目的 研究肠道补充益生元对急性胰腺炎肠上皮细胞紧密连接和屏障功能的影响.方法 Wistar大鼠腹腔注射左精氨酸建立AP模型,按随机数字法分组法分成对照组、AP组、益生元组.益生元组于造模前7 d开始给予益生元4 g·kg-1·d-1.建模后12 h处死大鼠,心脏取血检测血浆二胺氧化酶(DAO)活性,观察空肠病理变化、采用免疫组化方法检测紧密结合蛋白Occludin的含量.结果 AP组的血浆DAO活性为(7.29±0.68)U/L,显著高于对照组的(2.01±0.34)U/L(P＜0.01)和益生元组的(3.44±0.59)U/L(P＜0.05).AP组肠上皮occludin蛋白表达的灰度值为95.1±9.2,明显高于对照组的44.7±8.2和益生元组的59.7±7.8(P＜0.01).益生元组occludin表达也显著高于对照组(P＜0.05).结论 AP大鼠补充益生菌可增加肠上皮occludin蛋白表达,维持肠上皮紧密连接,维持正常的肠道通透性,从而达到保护肠黏膜屏障的效果.     

Early prediction of intestinal mucosal barrier function impairment by elevated serum procalcitonin in rats with severe acute pancreatitis

ObjectivesThe aim of this study was to evaluate serum procalcitonin (PCT) levels as a prognostic indicator of intestinal barrier function impairment in rats with severe acute pancreatitis (SAP).MethodsThirty-six male Sprague Dawley rats were randomly grouped into SAP group (injected sodium taurocholate via biliopancreatic duct), Gln group (gavaged with glutamine after modeling), and control group. Blood, pancreatic, and terminal ileum tissues were obtained from the rats after 6 h of modeling. Serum amylase (Amy) levels were determined using an automatic biochemical detector, while endotoxin (ET), diamine oxidase (DAO), and PCT levels were measured by ELISA test. The pathology of pancreatic and small intestine tissues were observed. PCT protein expression in intestinal tissues were detected by immunohistochemistry and western blot.ResultPancreatic and intestinal injuries in Gln group were significantly lower than SAP group. Serum amylase, DAO, and PCT levels in SAP and Gln groups differed greatly and were significantly higher than control group. Immuno-histochemistry and western blot results showed that PCT protein expression levels in small intestine tissues of SAP group were higher than Gln group and control group. Serum PCT levels had a significant correlation with serum endotoxin, DAO levels and intestinal mucosal injury scores.ConclusionPCT expression in serum and intestinal tissues in SAP rats increased significantly in the early stages of SAP, and was closely related to the onset and degree of intestinal barrier function impairment. Thus, our results showed that measuring serum PCT can be used to predict intestinal mucosal barrier function impairment in SAP rats.     

紧密连接蛋白occludin在急性坏死性胰腺炎大鼠脑微血管内皮细胞的表达

目的 研究ANP大鼠脑微血管内皮细胞紧密连接蛋白occludin表达的变化及与胰腺病变程度的关系.方法 80只大鼠按完全随机法分为假手术(SO)组和ANP 3、6、12、24 h组.以5%胆酸钠逆行性胰胆管注射诱导ANP大鼠动物模型,行胰腺组织病理学评价,应用免疫组织化学法、RT-PCR法及Western blotting法检测大鼠脑微血管内皮细胞间紧密连接蛋白occludin及其mRNA表达.结果 occludin蛋白沿脑血管线性表达.ANP 3 h组蛋白表达量从S0组的0.49±0.08下降到0.35±0.09;occludin mRNA表达从1.50±0.30减少到1.01±0.18(P<0.05).ANP 6 h组达最低值,分别为0.26±0.07和0.93±0.19.ANP 12、24 h组occludin蛋白和mRNA表达量又较ANP 6 h组增加(P<0.05),但仍低于SO组(P<0.05).occludin蛋白及mRNA表达与胰腺组织损害程度呈明显负相关(Pearson相关系数分别为-0.48和-0.536,P<0.05).结论 在ANP进程中,脑微血管内皮细胞间紧密连接蛋白occludin及其mRNA均呈下调趋势,且与胰腺病变程度呈负相关.     

A simple taurocholate-induced model of severe acute pancreatitis in rats

AIM: To investigate gut barrier damage and intestinal bacteria translocation in severe acute pancreatitis (SAP),a simple rat model of SAP was induced and studied.METHODS: Pancreatitis was induced by uniformly distributed injection of 3.8% Na taurocholate (1 mL/kg) beneath the pancreatic capsule. Rats in the control group were injected with normal saline in the identical location.RESULTS: Serum amylase, plasma endotoxin, intestinal permeability, and pancreatitis pathology scores were all markedly higher in the pancreatitis group than in the control group ( P < 0.01). The bacterial infection rate was significantly higher in the SAP group than in the control group ( P < 0.01), observed in parallel by both bacterial culture and real-time polymerase chain reaction. Acute damage of the pancreas was observed histologically in SAP rats, showing interstitial edema, leukocyte infiltration, acinar cell necrosis and hemorrhage. The microstructure of the intestinal mucosa of SAP rats appeared to be destroyed with loose, shortened microvilli and rupture of the intercellular junction, as shown by electron microscopy.CONCLUSION: Significant gut barrier damage and intestinal bacterial translocation were definitely observed with few potential study confounders in this SAP rat model, suggesting that it may be an appropriate animal model for study of gut barrier damage and bacterial translocation in SAP.     

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.     

RANTES在急性胰腺炎血-脑脊液屏障通透性变化中的作用

目的 观察急性胰腺炎(AP)大鼠脑组织正常T细胞表达与分泌的活化调节因子(regulated on activation,normal T-cell expressed and secreted,RANTES)和occludin在大脑中的表达,探讨RANTES表达与血-脑脊液屏障通透性变化之间的关系.方法 将49只SD大鼠按数字表法随机分为假手术(SO)组,急性水肿性胰腺炎(AEP)3、6 h组和急性坏死性胰腺炎(ANP)3、6、12、24 h组.以0.5%和5%牛磺胆酸钠逆行胰胆管注射制备AEP和ANP模型.采用RT-PCR、Western blotting和免疫组化法检测脑组织RANTES和occludin的mRNA及蛋白的表达.结果 SO组,AEP 3、6组,ANP 3、6、12、24 h组RANTES mRNA表达量分别为0、0、0、0.36±0.05、0.47 4-0.04、0.65 4-0.05、0.83±0.07,蛋白表达量为0、0、0、0.42±0.03、0.57±0.04、0.78±0.08、1.05±0.08,ANP组较SO组和AEP组显著增加(P<0.01);occludin mRNA表达量为1.21±0.07、1.17±0.07、1.15±0.08、0.84±0.07、0.77±0.05、0.64±0.09、0.56±0.09,蛋白表达量为1.18±0.08、1.16±0.10、1.11±0.10、0.90±0.03、0.65±0.05、0.57±0.05、0.48±0.05,ANP组较s0组和AEP组显著下降(P<0.01).RANTES表达与胰腺组织病理损伤呈正相关(r=0.936,P<0.001);occludin表达与胰腺组织病理损伤呈负相关(r=-0.900,P<0.001);RANTES和occludin呈负相关(r=-0.943,P<0.001).结论 ANP时大鼠脑组织RNANTES 表达呈进行性增高,occludin表达呈进行性下降,RANTES通过下调occludin蛋白表达而改变血-脑脊液屏障通透性.     

Influence of dexamethasone on mesenteric lymph node of rats with severe acute pancreatitis

AIM： To study the influence and mechanisms of dexamethasone on mesenteric lymph node of rats with severe acute pancreatitis （SAP）.
METHODS： The SAP rats were assigned to model, treated or sham-operated groups. The mortality, pathological changes of mesenteric lymph nodes, expression levels of NF-kB, P-selectin, Bax, Bcl-2 and caspase-3 protein and changes in apoptotic indexes in lymph nodes were observed at 3, 6 and 12 h after operation. The blood levels of endotoxin, superoxide dismutase （SOD）, malondialdehyde （MDA）, and endothelin-1 （ET-1） in blood were determined.
RESULTS： SOD content, expression of Bax protein and apoptotic index were significantly higher in the treated group than in the model group at different time points （P 〈 0.05 or P 〈 0.01）. Other blood-detecting indexes and histopathological scores of mesenteric lymph nodes were lower in the treated than in the model group （P 〈 0.05, P 〈 0.01 or P 〈 0.01）. NF-kB protein expression was negative in all groups. Comparing P-selectin and caspase-3 expression levels among all three groups, there was no marked difference between the model and treated group.
CONCLUSION： Dexamethasone can protect mesen-teric lymph nodes. The mechanism may be by reducing the content of inflammatory mediators in the blood and inducing lymphocyte apoptosis.     

Effects of gut barrier dysfunction and NF‐κB activation on aggravating mechanism of severe acute pancreatitis

OBJECTIVE: To study the effects of gut‐derived endotoxin translocation and NF‐κB activation on the aggravating mechanism of severe acute pancreatitis (SAP) and of treatment with pyrrolidine dithiocarbamate (PDTC) on rats with SAP. METHODS: SD rats were randomly divided into sham operation group (SO), SAP group, SAP + lipopolysaccharide(LPS) group, pyrrolidine dithiocarbamate (PDTC) treatment group and LPS group. Biochemical parameters and cytokines were examined in the serum. Multiple organs pathological slices were examined. Expression of NF‐κB mRNA in the liver tissue was detected by RT‐PCR. Activation of NF‐κB by the method of streptomycin avidin‐peroxidase (SP) and expression of NF‐κB p65 protein and its binding activity were analyzed by Western blot and electrophoretic mobidity shift assay (EMSA). RESULTS: Compared with sham operation group, the concentration of TNF‐α, alanine aminotransferase (ALT), and diamine oxidase (DAO) in serum significantly increased in SAP + LPS group (P < 0.05). Pathological changes were markedly observed in tissues and the expression of NF‐κB mRNA in the liver significantly increased (P < 0.05) also, the activation of NF‐κB and binding activity of NF‐κB p65 protein in the liver markedly increased (P < 0.01) in SAP + LPS group. Treatment with PDTC markedly reduced concentration of ALT, DAO and TNF‐α, and the expression of NF‐κB, and the pathologic scores, as well as significantly decreased the expression of NF‐κB p65 protein. CONCLUSION: The activation and overexpression of NF‐κB may participate in the aggravating mechanism of SAP. Treatment with PDTC has a protective effect on multiple organs damage in SAP.     

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。