内皮祖细胞移植对肺损伤炎症状态的影响

背景：内皮祖细胞在维持内皮系统功能及血管损伤后的修复中起重要作用,已广泛应用到心血管、下肢缺血、血管修复等多种疾病,但在炎症疾病及肺损伤中的研究较少。 目的：观察内皮祖细胞移植对急性肺损伤/急性呼吸窘迫综合征肿瘤坏死因子α及白细胞介素10的影响,探讨细胞移植能否改善急性肺损伤/急性呼吸窘迫综合征的炎症状态。 方法：同遗传背景SD大鼠30只随机均分为正常对照组、肺损伤组、细胞移植组。采用密度梯度离心法分离、培养SD大鼠的骨髓内皮祖细胞。肺损伤组、细胞移植组大鼠经尾静脉注射脂多糖建立急性肺损伤模型;正常对照组仅给予等量的磷酸盐缓冲溶液。造模半小时后,正常对照组、细胞移植组大鼠经尾静脉注入内皮祖细胞悬液;肺损伤组大鼠同法注入等量的磷酸盐缓冲溶液。 结果与结论：与肺损伤组相比,细胞移植组中白细胞介素10的水平显著增加(P < 0.001),肿瘤坏死因子α的表达下调,但差异无显著性意义(P > 0.05)。细胞移植促进白细胞介素10的表达,下调肿瘤坏死因子α的表达,能改善损伤肺组织的炎症状态。     

一种效果确切的大鼠重症急性胰腺炎模型

背景：目前尚无任何一种重症急性胰腺炎动物模型能与人类重症急性胰腺炎的发病过程完全一致,而用于实验治疗性研究的模型更是少之又少。 目的：建立一种大鼠重症急性胰腺炎模型,为进行大鼠重症急性胰腺炎的治疗性研究提供前提条件。 方法：30只SD大鼠随机分为对照组和模型组,对照组不建模,模型组大鼠采用逆行胰管穿刺注射法建立重症急性胰腺炎模型,建模后6,12,24 h,测血、腹水淀粉酶、血白细胞数量、血清白细胞介素6、肿瘤坏死因子α表达,并对胰腺组织进行病理评分。 结果与结论：模型组大鼠存活率80%,对照组大鼠存活率100%;模型组大鼠随时间的延长,病理评分、血、腹水淀粉酶、白细胞、白细胞介素6、肿瘤坏死因子α逐渐升高(P < 0.05)。说明逆行胰管注射法能够成功建立大鼠重症急性胰腺炎模型。     

脐带沃顿胶干细胞移植对脊髓损伤大鼠炎症因子表达的影响

背景：免疫炎症反应促使大量炎症因子的产生和释放是继发性脊髓损伤的其对主要原因。 目的：探讨脐带沃顿胶干细胞移植对急性脊髓损伤大鼠的神经修复作用及其对炎症因子单核细胞趋化蛋白1和白细胞介素10表达的影响。 方法：81只健康成年雄性SD大鼠,随机分为假手术组、模型组和脐带沃顿胶干细胞移植组(n=27),后两组以脊髓半切方法建立急性脊髓损伤模型,造模后脐带沃顿胶干细胞移植组经尾静脉移植1×10⁶个脐带沃顿胶干细胞。移植后不同时间通过BBB评分评价各组大鼠的运动功能;ELISA检测不同时间点各组大鼠血清中单核细胞趋化蛋白1的含量;qRT-PCR和Western分析不同时间点损伤脊髓组织中单核细胞趋化蛋白1及白细胞介素10的表达;免疫组织化学检测损伤组织中脐带沃顿胶干细胞的迁移和神经分化情况。 结果与结论：与假手术组和模型组相比,脐带沃顿胶干细胞移植组大鼠的神经功能明显恢复(P < 0.05)。脐带沃顿胶干细胞移植组大鼠血清中单核细胞趋化蛋白1水平显著低于模型组(P < 0.05)。脐带沃顿胶干细胞移植组脊髓组织单核细胞趋化蛋白1 mRNA和蛋白表达显著低于模型组(P < 0.05),白细胞介素10 mRNA和蛋白表达显著高于模型组(P < 0.05)。移植组中脐带沃顿胶干细胞可迁移至损伤部位,并表达神经胶质纤维酸性蛋白。这些结果提示脐带沃顿胶干细胞可能通过调控脊髓损伤组织的炎症反应,促进神经修复,这也可能是脐带沃顿胶干细胞治疗脊髓损伤的机制之一。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

青蒿琥酯对大鼠小肠移植急性排斥反应的作用

背景：随着强效、特异免疫抑制剂的问世,小肠移植的成活率有了一定程度的提高。但这些免疫抑制剂的不良反应及昂贵的治疗费用,让很多患者难以承受。所以,选择有免疫抑制作用的中药在临床上应用是很有意义的。青蒿琥酯有免疫抑制作用,可以减轻小肠移植急性排斥反应,提高小肠移植的成功率。目的：观察青蒿琥酯在大鼠小肠移植急性排斥反应中的作用及其机制。方法：选用封闭群SD大鼠和Wistar大鼠建立同种异基因小肠移植模型,随机分为3组：①同基因移植组(SD→SD)。②异基因移植组(Wistar→SD)。③异基因移植+青蒿琥酯治疗组(Wistar→SD+青蒿琥酯60 mg/(kg•d),腹腔注射)。结果与结论：同基因移植组大鼠存活均超过10 d,并于第10天全部处死。异基因移植组大鼠平均存活(6.73±0.58) d,治疗组大鼠平均存活(8.50±0.74) d,两组间比较差异有显著性意义(P < 0.01)。病理组织学检查显示同基因移植组标本无明显排斥征象,异基因移植组标本在术后第3,5,7天分别符合轻、中、重度排斥反应,治疗组部分标本有轻度排斥表现,但出现较晚、程度较轻。ELISA法检测显示异基因移植组血清白细胞介素2、γ-干扰素表达水平在术后均显著高于其他2组(P < 0.01),治疗组血清白细胞介素2表达水平与同基因移植组比较亦有升高,但差异无显著性意义(P > 0.05),治疗组血清γ-干扰素表达水平高于同基因移植组(P < 0.05)。结果可见青蒿琥酯对大鼠小肠移植急性排斥反应有抑制作用,其作用机制可能与抑制白细胞介素2、γ-干扰素等细胞因子的分泌表达有关。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

骨髓间充质干细胞静脉移植脊髓损伤大鼠肿瘤坏死因子α及 白细胞介素1β的表达

背景：研究认为间充质干细胞的营养支持在脊髓损伤治疗中起了主要作用,其同损伤宿主神经组织间的相互作用可导致一些不利于损伤修复的炎症因子表达减少。 目的：观察大鼠骨髓间充质干细胞静脉注射移植对脊髓损伤后肿瘤坏死因子α、白细胞介素1β 表达的影响。 方法：运用改良Allen法制备大鼠T10脊髓外伤性截瘫模型,随机分为对照组和骨髓间充质干细胞移植组,设未损伤脊髓的假手术组做对照。骨髓间充质干细胞移植组、假手术组均接受大鼠骨髓间充质干细胞静脉注射移植,对照组静脉注射等量PBS。 结果与结论：对照组和骨髓间充质干细胞移植组损伤脊髓肿瘤坏死因子α、白细胞介素1β蛋白表达较假手术组有明显增加(P < 0.05);骨髓间充质干细胞移植组与对照组比较, 肿瘤坏死因子α、白细胞介素1β蛋白表达受到明显抑制(P < 0.05)。提示大鼠骨髓间充质干细胞静脉移植后能使损伤脊髓局部的肿瘤坏死因子α、白细胞介素1β表达程度降低。这可能是改变脊髓损伤区的微环境,减少脊髓继发性损伤,促进损伤大鼠运动功能恢复的机制之一。     

人脐血单个核细胞静脉移植联合培哚普利对急性心肌梗死炎症反应的影响

背景：近期报道骨髓间充质干细胞心肌内直接移植联合培哚普利治疗急性心肌梗死大鼠,可改善心肌组织内环境,并增强急性心肌梗死疗效。 目的：观察人脐血单个核细胞静脉移植联合血管紧张素转化酶抑制剂培哚普利对家兔急性心肌梗死心肌组织炎症反应与促炎因子白细胞介素6表达及心功能影响,并探讨联合治疗对急性心肌梗死可能的保护机制。 方法：人脐血单个核细胞取自健康足月分娩产妇脐血。60只健康家兔制备急性心肌梗死模型,建模成功后随机数字表法均分对照组、培哚普利组、单纯移植组和联合治疗组。每组随机选5只家兔分别于移植后1,2,4周超声心动图检测家兔心功能指标左室射血分数及左室短轴缩短率;苏木精-伊红染色光镜观察心肌病理变化和白细胞计数;免疫组化检测心肌组织白细胞介素6蛋白表达量;荧光显微镜观测绿色荧光蛋白阳性细胞。 结果与结论：①与对照组比较,培哚普利组、单纯移植组、联合治疗组治疗后1,2,4周心功能指标左室短轴缩短率及左室射血分数改善(P < 0.05),单纯移植组高于培哚普利组(P < 0.05),联合治疗组改善最显著(P < 0.05)。②与对照组比较,培哚普利组、单纯移植组、联合治疗组治疗后1,2,4周心肌组织白细胞计数及白细胞介素6的表达均显著减低(P < 0.05),且单纯移植组低于培哚普利组(P < 0.05),联合治疗组最低(P < 0.05)。③联合治疗组、单纯移植组治疗后1,2,4周均可见绿色荧光蛋白阳性细胞散在分布于梗死周边区域,且联合治疗组细胞计数多于单纯移植组(P < 0.05)。说明培哚普利联合人脐血单个核细胞静脉移植治疗急性心肌梗死实验动物,能提高移植细胞在心肌组织内存活率,并进一步改善心功能。其机制可能与联合治疗抑制心肌局部炎症反应及促炎因子白细胞介素6水平表达作用增强有关。     

虾青素对缺氧缺血性脑损伤新生大鼠模型的神经保护作用

背景：研究发现虾青素有良好的神经保护作用,但是对于其在新生儿缺氧缺血性损伤中的治疗作用,目前尚无相关报道。目的：构建缺氧缺血性脑损伤新生大鼠模型,观察虾青素对其产生的神经保护作用及作用的途径。方法：从98只7 d龄的SD乳鼠中随机取30只作为假手术组,其余大鼠结扎左颈总动脉2 h后,置于体积分数92%的特种标准气体、8%的氧气缺氧舱2 h建立缺血缺氧性脑损伤模型。假手术组仅分离颈总动脉,不予缺血缺氧处理。将造模成功的大鼠随机分为脑缺血缺氧组和虾青素治疗组,各30只。虾青素治疗组大鼠在脑缺血缺氧模型建成后立即通过腹腔注射80 mg/kg虾青素。结果与结论：与假手术组相比,脑缺血缺氧组大鼠缺血损伤区顶叶皮质中p-Akt、p-GSK3β、cleaved-caspase3蛋白的表达水平显著增加,Bcl-2蛋白的表达水平显著减少(P < 0.05);与脑缺血缺氧组相比,虾青素治疗可以显著减少凋亡相关蛋白cleaved-caspase3蛋白的表达水平(P < 0.05),显著上调Bcl-2蛋白的表达水平(P < 0.05),明显减少凋亡细胞的数量(P < 0.05)。提示虾青素可以显著改善新生大鼠缺氧缺血性脑损伤的预后及作用途径与上调Akt/GSK3β信号通路相关。中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

无创伤双后肢缺血后处理对移植胰腺缺血再灌注损伤的远隔保护

背景：缺血预处理及缺血后处理是近年来提出减轻缺血再灌注损伤有效方法。 目的：探讨无创伤双后肢缺血后处理对移植胰腺缺血再灌注损伤的影响及机制。 方法：18只糖尿病SD大鼠数字表法随机分为3组,对照组仅行开腹术;缺血再灌注组仅行胰腺移植;缺血后处理组,移植前行非创伤性双后肢缺血后处理。 结果与结论：缺血再灌注组血糖和胰腺组织中丙二醛水平均高于缺血后处理组(P < 0.01)、而超氧化物歧化酶活性低于缺血后处理组(P < 0.01);与缺血后处理组比较,缺血再灌注组胰腺组织凋亡指数明显增高(P < 0.01)。结果提示,无创伤双后肢缺血后处理对大鼠移植胰的缺血再灌注损伤具有保护作用,机制可能与可通过减少超氧化物歧化酶失活,从而清除氧自由基以及减少胰腺细胞凋亡等有关。     

急性肌肉软组织损伤后不同冷疗方式处理的组织学变化

背景：冷疗处理急性软组织损伤已在临床广泛应用。 目的：观察不同冷疗方式对急性软组织损伤大鼠的组织学改变及治疗效果。 方法：将新生Wistar大鼠随机分成正常组、模型组、间断冷敷组及持续冷敷组,后3组建立急性软组织损伤动物模型。间断冷敷组用4 ℃生物冰袋间断冷敷于损伤部位,持续冷敷组用4 ℃生物冰袋持续冷敷,模型组不予以处理。冷敷48 h后观察各组损伤部位大体形态改变,采用损伤症候指数评估损伤程度。 结果与结论：与模型组比较,间断冷敷组及持续冷敷组损伤症候指数与组织学评分较低,白细胞介素1β的阳性表达率降低,转化生长因子β1的阳性表达率表达率升高  (P < 0.05)。与间断冷敷组比较,持续冷敷组损伤症候指数与组织学评分较低(P< 0.05),白细胞介素1β的阳性表达率降低(P< 0.05),转化生长因子β1的阳性表达率表达率升高(P < 0.05)。结果证实,冷疗处理治疗急性期软组织损伤的机制与降低白细胞介素1β及提高转化生长因子β1表达有关,持续冷疗的疗效优于间断冷疗。     

N-乙酰半胱氨酸对烟熏模型大鼠肺组织氧化应激的影响

背景:N-乙酰半胱氨酸是谷胱甘肽的前体,可以直接清除氧自由基。然而,N-乙酰半胱氨酸是否通过降低氧化应激反应,减轻吸烟导致的肺损伤尚不完全清楚。目的:探讨N-乙酰半胱氨酸对烟熏大鼠肺组织氧化应激的影响,并阐明其可能的作用机制。方法:将30只雄性大鼠随机分为对照组、烟熏组、烟熏+N-乙酰半胱氨酸组。将烟熏组和烟熏+N-乙酰半胱氨酸组大鼠置于被动吸烟动物染毒系统,每次暴露20支香烟的烟雾中,2次/d,1 h/次,持续8周。N-乙酰半胱氨酸组大鼠每天被动烟熏前给予N-乙酰半胱氨酸200 mg/kg灌胃,持续8周。正常对照组大鼠仅单纯放置在染毒系统内,正常饲养8周。实验方案由沈阳医学院实验动物伦理委员会于2018年10月批准,批准号:研伦审第(2018)85号。结果与结论:①病理观察发现烟熏组大鼠肺组织排列紊乱、肺泡间隔增厚、炎症细胞浸润和间质纤维化,而烟熏+N-乙酰半胱氨酸组肺组织出血、间质内炎症细胞数量显著减少、纤维化程度减轻;②与对照组相比,烟熏组大鼠肺组织丙二醛5和肌醇酶α基因表达升高,超氧化物歧化酶1基因表达显著降低,而N-乙酰半胱氨酸可抑制上述变化;③免疫荧光和Western blot结果均发现烟熏+N-乙酰半胱氨酸组大鼠肺组织丙二醛5和肌醇酶α蛋白表达较烟熏组显著降低,超氧化物歧化酶1蛋白表达较烟熏组升高。此外,烟熏+N-乙酰半胱氨酸组核因子E2相关因子和Keap1蛋白表达明显升高,Bach1蛋白表达显著降低;④结果表明,N-乙酰半胱氨酸通过降低氧化应激对烟熏导致的肺损伤发挥保护作用,其作用机制可能通过激活核因子E2相关因子/Keap1信号通路实现。     

N-乙酰半胱氨酸对重症急性胰腺炎大鼠肺损伤作用的实验研究

目的:探讨N-乙酰半胱氨酸(NAC)对重症急性胰腺炎(SAP)大鼠肺损伤的作用。方法:雄性SD大鼠30只,随机分为假手术组(SO组)、SAP组、NAC组。胆胰管逆行注射5%牛磺胆酸钠制备SAP模型,造模后30min腹腔注射5%NAC(0.2ml/100g)干预SAP模型,12h后处死大鼠。检测各组血清淀粉酶(AMY)、肺组织髓过氧化物酶(MPO)、胰腺和肺组织病理学评分、肺组织肿瘤坏死因子-α(TNF-α)和细胞间粘附分子-1(ICAM-1)mRNA表达的变化。结果:与SO组比较,SAP组AMY、MPO、胰腺和肺组织病理学评分明显升高(P<0.01),TNF-α和ICAM-1mRNA表达明显增强(P<0.01);应用NAC处理后,AMY和MPO水平下降,胰腺和肺组织损伤缓解,TNF-α和ICAM-1mRNA表达减弱,与SAP组有明显差异(P<0.01)。结论:NAC对SAP大鼠肺损伤具有保护作用,其机制可能与抑制肺组织TNF-α和ICAM-1mRNA的表达有关。     

Effect of Emodin on Endoplasmic Reticulum Stress in Rats with Severe Acute Pancreatitis

This study aimed to investigate the protective effect of emodin on endoplasmic reticulum (ER) stress in rats with severe acute pancreatitis (SAP) and the underlying molecular mechanism. Sprague–Dawley male rats were randomly divided into sham operation group, SAP model group, and emodin treatment group. SAP was constructed through injecting sodium taurocholate into pancreatic and biliary duct in rats. Half an hour before establishing the animal model, emodin or sodium carboxymethylcellulose was intragastrically administrated to the rats in respective group. Rats were killed at 3, 6, and 12 h postdisease induction. The amylase, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) levels in serum, pancreatic histopathology, acinar ER ultrastructure, protein expression of Bip, IRE1α,TRAF2, ASK1, p-JNK, and p-p38 MAPK in pancreas were examined. Sodium taurocholate induced pancreatic injury and ER lumen dilated in exocrine pancreas in rats at 3-, 6-, and 12-h time points. ER stress transducers Bip, IRE1α, and their downstream molecules TRAF2, ASK1 in pancreatitis were upregulated. Furthermore, phosphorylation of JNK and p38MAPK in pancreas was increased, which induced high expression level of inflammatory cytokines such as TNF-α and IL-6. Treatment with emodin obviously ameliorated pancreatic injury and decreased the release of amylase and inflammatory cytokines. Further studies showed that emodin significantly decreased the expression of Bip, IRE1α, TRAF2, and ASK1, inhibited phosphorylation of JNK and p38 MAPK in pancreas in rats at all time points. Emodin could reduce pancreatic injury and restrain inflammatory reaction in SAP rats partly via inhibiting ER stress transducers IRE1α and its downstream molecules.     

应激在急性出血坏死性胰腺炎所致肺损伤中的作用

目的:通过复制大鼠急性出血坏死性胰腺炎(AHNP)模型,探讨在AHNP的发病过程中,应激与AHNP及其所致肺损伤的关系。方法:用5%牛磺胆酸钠溶液逆行胰胆管注射制作大鼠AHNP模型。将实验动物随机分为A-C3组,A组为假手术组,B组为AHNP组,C组在制成AHNP模型后给予甲吡酮(metyrapone)以控制应激反应的程度,观察大鼠血清中皮质醇,C反应蛋白(CRP)及血清淀粉酶的含量,以及肺及胰腺的病理组织学。结果:C组动物血清中皮质醇、CRP及淀粉酶的含量明显低于对照组,肺及胰腺的损害程度,明显低于对照组(P＜0.05)。结论:应激在急性出血坏死性胰腺炎并发肺损伤的过程中起了重要作用。运用特异性抗应激药物可以明显减轻其所致的肺损伤。     

不同剂量花姜酮对大鼠重症急性胰腺炎的影响

目的:探讨花姜酮( Zerumbone)静脉给药对大鼠重症急性胰腺炎(SAP)的治疗作用。方法:雄性Wistar大鼠50只,随机分为5组:假手术组(SO组)、SAP模型组(SAP组)、5mg/kg花姜酮预处理组(5mg/kg花姜酮组)、10mg/kg花姜酮预处理组(10mg/kg花姜酮组)、20 mg/kg花姜酮预处理组(20mg/kg花姜酮组)。胆胰管逆行注射5%牛磺胆酸钠制备SAP模型。SO组、SAP组造模前30 min股静脉注射10%二甲基亚砜(DMSO)(2ml/kg),余三组注射等量10%DMSO溶解的等体积、不同浓度花姜酮。术后12h心脏取血检测各组血清淀粉酶(AMY),谷氨酸氨基转移酶(ALT),血肌酐(Cr)浓度变化,并观察胰腺组织病理学改变。结果:与SAP组比较,5 mg/kg花姜酮组AMY和胰腺病理评分没有改变,ALT和Cr有改善(P0.05);10mg/kg花姜酮组上述指标较SAP组显著降低(P0.05);20mg/kg花姜酮组AMY、Cr和胰腺组织病理学评分较SAP组显著降低(P0.05),但ALT与SAP组比较差异无统计学意义(P0.05)。结论:花姜酮治疗大鼠SAP,以10 mg/kg静脉注射比较安全、有效。     

骨髓间充质干细胞移植治疗重症急性胰腺炎肺损伤

BACKGROUND:A large number of studies have confirmed that bone marrow mesenchymal stem cells can couple with the circulation of the blood to other organs, promote pancreatic tissue repair injury and reduce pulmonary fibrosis, which have certain therapeutic effects on pancreas and lung injuries. OBJECTIVE:To study the therapeutic effect on severe acute pancreatitis-associated lung injury in rats after the transplantation of bone marrow mesenchymal stem cells. METHODS:Animal models of severe acute pancreatitis-associated lung injury were prepared in rats via retrograde injection of 4% sodium taurocholate. Sprague-Dawley rats were randomized into three groups and received bone marrow mesencnymal stem cell injection via the tail vein in transplantation group, the same volume of normal saline in control group, or no treatment in normal groups. All the treatments in each group were performed 24 hours after modeling. Twenty-four hours after transplantation, hematoxylin-eosin staining of the pancreatic and lung tissues was performed. mRNA expressions of tumor necrosis factor-α and interleukin-1β in pancreatic and lung tissues were detected. ELISA kit was used to detect levels of serum C-reactive protein and tumor necrosis factor-α. RESULTS AND CONCLUSION:After modeling, under hematoxylin-eosin staining, there were a large number of inflammatory cells infiltrating in the damaged pancreatic tissues, accompanied by incomplete acinar structures, seriously destroyed lobular structures, alveolar fusion in the lung tissues, thickening of the alveolar walls, and a large amount of inflammatory cells infiltrating in the alveoli. These findings indicated successful modeling of severe acute pancreatitis-associated lung injury in rats. After cell transplantation, the number of infiltrated inflammatory cells in the damaged pancreatic tissue was reduced, with clear lobular structures and no bleeding from the acini; the structure of lung tissues was clear, with complete alveolar walls, and the width of alveolar space was reduced. Immunohistochemical results showed that transplanted DAPI-labeled bone marrow mesenchymal stem cells were aggregated in the pancreas and lung tissue, and uneven distributed in the damaged area. No DAPI expression in the pancreas and lung tissue was found in the control group, indicating transplanted bone marrow mesenchymal stem cells migrated into the damaged pancreas and lung tissue through the blood circulation, to further repair the damage area. RT-PCR test results showed that compared with the control group, bone marrow mesenchymal stem cell transplantation significantly reduced the levels of tumor necrosis factor-α and interleukin-1β in the pancreatic and lung tissues (P < 0.05). Higher levels of C-reactive protein and tumor necrosis factor-α were found in the control group compared with the normal group (P < 0.01), while the lower levels were obtained in the control group (P < 0.05). To conclude, our findings suggest that bone marrow mesenchymal stem cell transplantation is an effective therapy for severe acute pancreatitis-associated lung injury, and its mechanism may be associated with the reduction of inflammatory reactions and translation into the pancreas and lung tissue.     

Role of neutrophils in the activation of trypsinogen in severe acute pancreatitis

The relationship between inflammation and proteolytic activation in pancreatitis is an unresolved issue in pancreatology. The purpose of this study was to define the influence of neutrophils on trypsinogen activation in severe AP. Pancreatitis was induced by infusion of taurocholate into the pancreatic duct in C57BL/6 mice. For neutrophil depletion, an anti-Gr-1 antibody was administered before pancreatitis induction. Administration of the anti-Gr-1 antibody reduced circulating neutrophils by 97%. Pancreatic TAP and serum amylase levels increased 2 h and 24 h after induction of pancreatitis. Neutrophil depletion reduced pancreatic TAP and serum amylase levels at 24 h but not at 2 h after pancreatitis induction. Pancreatic MPO and infiltration of neutrophils, as well as MIP-2 levels, were increased 24 h after taurocholate infusion. Two hours after taurocholate administration, no significant pancreatic infiltration of neutrophils was observed. Injection of the anti-Gr-1 antibody abolished MPO activity, neutrophil accumulation, and MIP-2 levels, as well as acinar cell necrosis, hemorrhage, and edema in the pancreas at 24 h. Moreover, taurocholate-provoked tissue damage and MPO activity in the lung were normalized by neutrophil depletion. Intravital fluorescence microscopy revealed a 97% reduction of leukocytes in the pancreatic microcirculation after administration of the anti-Gr-1 antibody. Our data demonstrate that initial trypsinogen activation is independent of neutrophils, whereas later activation is dependent on neutrophils in the pancreas. Neutrophils are critical in mediating pancreatic and lung tissue damage in severe AP.     

Oxidative stress is enhanced by hypothermia imposed on cerulein-induced pancreatitis in rats

BACKGROUND: Hypothermia is a frequent event in severe acute pancreatitis (AP) and its real effects on the normal pancreas have not been well demonstrated. Moreover, neither have its effects on the outcome of acute pancreatitis been fully investigated. One hypothesis is that oxidative stress may be implicated in lesions caused or treated by hypothermia. AIM OF THE STUDY: To investigate the effect of hypothermia in cerulein-induced acute pancreatitis (CIAP) in rats and the role played by oxidative stress in this process. METHODS: Male Wistar rats were divided into hypothermic and normothermic groups. Hypothermia was induced with a cold mattress and rectal temperature was kept at 30 masculineC for one hour. Acute pancreatitis was induced with 2 doses of cerulein (20 ìg/kg) administered at a one-hour interval. Serum amylase, pancreas vascular permeability by Evan's blue method, pancreas wet-to-dry weight ratio and histopathology were analyzed in each group. RESULTS: When compared with normothermic rats, hypothermic animals, with cerulein-induced acute pancreatitis, showed higher levels of pancreatic vascular permeability (p < 0.05), pancreas wet-to-dry weight ratio (p = 0.03), and histologically verified edema (p < 0.05), but similar serum amylase levels. The hypothermic group showed a higher oxidized-reduced glutathione ratio than the normothermic group. CONCLUSION: Moderate hypothermia produced a greater inflammatory response in established acute pancreatitis induced by cerulein in rats. Moreover, this study suggests that oxidative stress may be one of the mechanisms responsible for the worse outcome in hypothermic rats with cerulein-induced acute pancreatitis.     

Leukotriene Generation and Neutrophil Infiltration After Experimental Acute Pancreatitis

The role of 5-lipoxygenase metabolites of arachidonic acid in the inflammatory response associated with experimental acute pancreatitis has been evaluated. For this purpose, an experimental necrohemorrhagic pancreatitis was induced in rats by intraductal administration of 5% sodium taurocholate. Neutrophil infiltration was detected in pancreas at 1 and 3 h after the induction of pancreatitis. This was concomitant with increased levels of leukotriene B₄ and peptide leukotrienes (C₄, D₄ and E₄). In lung, similar increases in neutrophil infiltration were detected but only 3 h after acute pancreatitis induction, and no changes in leukotriene B₄ nor peptide leukotrienes were apparent at this time. These results suggest that after induction of acute pancreatitis, 5-lipoxygenase metabolites could play a role in the inflammatory response in the pancreas, but they are not involved in the inflammatory response in lung.     

Fas/FasL介导的caspase-3活化与急性胰腺炎腺泡细胞凋亡的关系

目的： 研究凋亡调控基因及蛋白Fas、FasL和caspase-3在大鼠急性胰腺炎（AP）组织中的表达及其相互关系。方法：经胰胆管逆行注射不同浓度的牛磺胆酸钠建立不同炎症程度的AP模型,采用RT-PCR、Western blotting技术检测大鼠胰腺炎组织Fas、FasL和caspase-3蛋白及mRNA的表达, TUNEL法检测胰腺炎组织腺泡细胞凋亡。结果：在正常胰腺组织内即可见Fas、FasL、caspase-3蛋白和mRNA的表达;建立AP模型后,随胰腺炎症程度的加重,Fas、FasL、caspase-3蛋白和mRNA的表达逐渐下降,腺泡细胞凋亡率亦逐渐下降,且caspase-3 表达水平在各个组间的变化趋势与Fas/FasL系统的变化趋势相一致。结论：Fas/FasL系统介导的凋亡途径参与了急性胰腺炎腺泡细胞凋亡的调节。     

Spatial and temporal differences of HMGB1 expression in the pancreas of rats with acute pancreatitis

We aimed to investigate the spatial and temporal differences in expression between HMGB1 and early-stage inflammatory cytokines (IL-1, IL-6 and TNF-α) in pancreas tissue in rats with acute pancreatitis. SD rats (BW 350 ± 30 g, n = 48) were randomly divided into the experimental group (n = 36) which were injected with 5% sodium taurocholate into the bilipancreatic duct retrogradely to produce acute necrotic pancreatitis (ANP) rat models, and the sham-operated (SO) group (n = 12) injected with equal dose of saline. The rats were sacrificed at different time points at 0 h, 3 h, 6 h, 12 h, and 24 h post modeling, respectively. The peripheral blood amylase and different inflammatory factors in ANP rats at different time points were detected by ELISA, and the expression of HMGB1 in the pancreatic tissue was detected by immunohistochemistry, Western blot and Q-PCR methods. Results showed that the serum amylase in the ANP model rats was significantly higher than the sham-operated group (P < 0.05). The early inflammatory factors (IL-1, TNF-α and IL-6) increased quickly at 3 h after the model induction, reached the peak level at 6 h (higher than SO group, P < 0.05), then decreased at 12 h, and at 24 h the levels were lower than those at 12 h (P < 0.05). The HMGB1 level in the pancreatitis tissue did not change significantly at 3 h and 6 h (P > 0.05), however, it increased remarkably at 12 h, and maintained up to 24 h (P > 0.05). As a late inflammatory factor, the expression of HMGB1 in acute pancreatitis was obviously later than the early inflammatory factors IL-1, TNF-α and IL-6. HMGB1 may play a key role in maintaining the development of the acute pancreatitis.