丹参素干预体外低温保存大鼠肝脏血红素氧合酶1的表达

背景：研究发现丹参能够降低诱导型一氧化氮合酶mRNA 的表达，减少一氧化氮的产生，抑制肿瘤坏死因子、白细胞介素等炎性因子的分泌，具有抗氧化作用。丹参素作为丹参的重要单体成分，是否具有相同的作用? 目的：验证丹参素干预大鼠肝脏血红素氧合酶1的表达，以及对体外肝脏低温保存肝脏的保护。 方法：建立大鼠肝脏低温灌注保存模型。分为3组，对照组术前腹腔注射生理盐水，术中用乳酸林格液灌注；实验组术前腹腔注射生理盐水，术中用丹参素+乳酸林格液灌注；抑制剂组：术前腹腔注射锌原朴啉，术中用丹参素+乳酸林格液灌注。保存0，1，3，6 h，分别RT-PCR检测血红素氧合酶1mRNA及蛋白表达的情况，检测细胞线粒体钙离子含量及钙离子ATP酶活性，电镜观察各组肝细胞、线粒体形态改变，光镜观察肝细胞、肝小叶形态改变。 结果与结论：实验组肝血红素氧合酶1 mRNA及蛋白的表达水平明显比其他两组高(P < 0.05)。实验组的肝细胞线粒体钙离子含量明显较其他两组低，Ca2+-ATP酶活性较其他两组高。结果提示，丹参素灌注保存液可以诱导大鼠肝脏中血红素氧合酶1的过表达，延长肝脏低温保存时间。     

迟发缺血预处理低温保存肾移植供体中血红素加氧酶1的表达

背景：缺血预适应延迟反应通过诱导保护性蛋白增强组织对缺血再灌注损伤的耐受能力；血红素加氧酶1参与缺血预适应延迟保护作用。迟发缺血预处理对低温保存肾脏的作用及血红素加氧酶1是否参与其中尚不清楚。 目的：观察缺血预处理诱导血红素加氧酶1的迟发缺血预处理反应对低温保存肾脏移植供体的作用。 方法：雄性SD大鼠随机分入5组：空白对照组、低温保存组、缺血预处理组、缺血+低温组(n=12)；缺血+给药+低温组。各组大鼠均行右肾切除，预处理或假手术操作处理后24 h采用大鼠肾脏非循环离体灌注模型获取肾脏，分别于保存24，48，72 h取样。缺血+给药+低温组除上述处理外，还于原位低温灌注术前1 h接受1次血红素加氧化酶1抑制剂锡原卟啉腹腔注射。低温保存肾脏于各保存终点留取保存液，测定pH值和乳酸脱氢酶含量；切取1/2肾脏按照光镜要求制备标本送检；剩余1/2肾脏用于免疫印迹法测定血红素加氧酶1表达，比色法测定皮质Na-K-ATP酶活性、丙二醛和还原型谷胱甘肽含量；未保存肾脏仅通过免疫印迹法测定血红素加氧酶1的基础表达情况。 结果与结论：迟发缺血预处理诱导了肾组织血红素加氧酶1的表达，与单纯低温保存组相比保存24，48 h后，缺血+低温组保存液pH值、乳酸脱氢酶活性降低；肾脏组织Na-K-ATP酶活性、谷胱甘肽含量增加，丙二醛含量降低；同时点预处理组肾组织光镜形态学改变稍好于单纯低温保存组。给予血红素加氧酶1抑制剂后，这种保护作用消失。提示，迟发缺血预处理延长了肾脏低温保存时限，这可能与诱导血红素加氧酶1，增加组织抗氧化能力，减轻低温保存氧应激有关。     

丹参对原位肝移植大鼠供肝冷保存再灌注损伤的保护作用☆

目的: 研究表明，丹参对心、脑、肝等重要器官的缺血再灌注损伤有保护作用。制备大鼠异体原位肝移植模型，验证丹参对大鼠肝移植缺血再灌流损伤的保护作用。 方法：实验于2006-10/2007-08在南方医院中心实验室及动物实验中心完成，动物实验方法符合动物伦理学要求。①实验材料及分组：选用SD大鼠40只，按随机数字表法分为假手术组、模型对照组和丹参注射液组，假手术组8只，模型对照组、丹参注射液组各8对（供体与受体）。②实验方法：建立原位肝移植模型，在供肝灌注冷保存时，以4 ℃ 乳酸林格氏液为基液，丹参注射液组灌注保存液中加60 mL/L丹参注射液；模型对照组不加丹参。③实验评估：移植术后6 h处死各组大鼠取样，检测血清谷草转氨酶、谷丙转氨酶及乳酸脱氢酶活性；测定肝组织中丙二醛含量及超氧化物歧化酶、谷胱甘肽过氧化物酶活性，并对比观察移植肝病理形态学改变。 结果：模型对照组和丹参注射液组16只受体大鼠及假手术组8只大鼠全部进入结果分析，无脱失。①丹参注射液组和模型对照组移植肝再灌注后血清谷丙转氨酶、谷草转氨酶及乳酸脱氢酶活性均高于假手术组(P < 0.01)；丹参注射液组低于模型对照组(P < 0.01)。②丹参注射液组肝组织中丙二醛含量较模型对照组明显下降(P < 0.01)，超氧化物歧化酶和谷胱甘肽过氧化物酶的活性则明显升高 (P < 0.01)。③丹参注射液组较模型对照组肝组织肝细胞坏死程度减轻，炎性细胞浸润减少，肝组织再灌注损害程度减轻。 结论：丹参对原位肝移植肝脏的缺血再灌注损伤有保护作用，从而减轻氧自由基及脂质过氧化，保护细胞膜，改善肝功能。     

缺血预处理减轻大鼠减体积肝移植损伤并促进肝再生

背景：近年来,肝移植技术迅速发展,如何预防缺血再灌注损伤并有效保护肝再生成为研究的热点。缺血预处理是保护肝缺血损伤的有效方法,但其确切机制尚存争议。 目的：研究缺血预处理在大鼠减体积肝移植肝损伤和肝再生中的作用及机制。 方法：动物随机分为3组,肝移植组建立大鼠减体积肝移植模型。缺血预处理+肝移植组在供肝灌注前阻断第1肝门行缺血预处理10 min,再灌注15 min。假手术组在开腹后游离肝周韧带,然后关腹。分别于术后0.5,2,6,24 h取材。通过血清谷丙转氨酶水平和移植肝组织病理检查评估肝损伤。半定量免疫组织化学和western blot法测定氧化还原蛋白1表达水平,检测移植肝细胞增殖细胞核抗原评估肝再生情况。 结果与结论：与肝移植组相比,缺血预处 理+肝移植组术后6,24 h受体血清谷丙转氨酶明显降低(P < 0.05;P < 0.01)。病理学分析显示肝移植组术后24 h可见到门脉周围大量炎细胞浸润,肝窦扩张明显,肝组织损伤较重;而缺血预处理+肝移植组则损伤较轻。半定量免疫组织化学显示缺血预处 理+肝移植组移植肝中Ref-1蛋白表达明显增加,这一结果同样在westernblot检测中得到验证：缺血预处理+肝移植组移植肝术后24 h Ref-1蛋白表达较肝移植组明显增强 (P < 0.05)。同时,术后2,6和24 h 缺血预处理+肝移植组增殖细胞核抗原阳性细胞数较肝移植组明显增加(P < 0.05)。结果提示缺血预处理可减轻大鼠减体积肝移植术后早期移植物肝损伤并促进肝再生,这与Ref-1蛋白高表达密切相关。     

自制多脏器保存液对大鼠睾丸一氧化氮合酶的影响***

背景：在睾丸移植过程中,低温保存和缺血可导致睾丸产生氧自由基而损伤睾丸组织。 目的：观察自制多脏器保存液对低温保存大鼠睾丸一氧化氮合酶的影响。 方法：采用自制多脏器保存液和UW液低温保存大鼠睾丸,分别于保存24,48,72 h时切取睾丸,测定睾丸组织内的总抗氧化能力和一氧化氮合酶活性。 结果与结论：自制多器官保存液低温保存各时点大鼠睾丸一氧化氮合酶活性和总抗氧化能力与UW液组比较差异均无显著性意义(P > 0.05)。表明自制多脏器保存液能明显减轻低温保存大鼠睾丸氧自由基损伤,其作用与经典的美国威斯康星大学UW保存液基本相当。     

自制PV液对大鼠肝脏低温保存的实验研究

【摘要】 目的 探讨自制PV液对大鼠肝脏低温保存的效果。方法 Wistar大鼠90只随机分成三组：UW液组、PV液组和生理盐水（NS）对照组，采用大鼠肝脏非循环离体灌注模型，每组对大鼠肝脏保存0h、6h、12h、18h、24h，每个亚组6只大鼠。测定保存不同时间段后肝脏酶学变化（ALT、AST、LDH）、灌注流出液中氧自由基代谢产物（MDA、SOD）的含量，观察胆汁分泌量及镜下肝脏形态学变化。结果 PV液组与UW液组比较，灌注流出液中ALT、AST、LDH、SOD含量相近，无显著差异（P>0.05）；保存6h后，PV液组TNF-α值较UW液组升高明显（P<0.05）；保存12h后UW液组MDA含量升高较PV液组明显（P<0.05)；保存18h后PV液组胆汁分泌量低于UW液组（P<0.05）；两组光镜、电镜下组织形态改变相似。结论 PV液与UW液对Wistar大鼠肝脏功能具有保护作用，两者短时间保存效果相似，在抗氧化及清除氧自由基方面，PV液略优于UW液。     

缺血预处理预防肝细胞分离过程中的缺血再灌注损伤

背景：缺血预处理能否减轻肝细胞分离过程中的缺血再灌注损伤及改善供体残留肝脏功能? 经检索国内外罕见这方面的研究。 目的：探讨缺血预处理对分离肝细胞及供鼠残肝缺血再灌注损伤的防治作用。 方法：12 只SD大鼠随机分为2组：单纯肝部分切除组和缺血预处理组,各6只。采用改良四步胶原酶灌注法分离上述切除肝脏肝细胞,同时收集术前和术后1 d大鼠的血清。 结果与结论：缺血预处理组切除肝脏分离肝细胞成活率、增殖活性及白蛋白合成、超氧化物歧化酶水平显著高于单纯肝部分切除组(P < 0.05),而乳酸脱氢酶、谷丙转氨酶、丙二醛水平显著减少(P < 0.05);与单纯肝部分切除组比较,缺血预处理组大鼠血清白蛋白、乳酸脱氢酶、谷丙转氨酶、超氧化物歧化酶、丙二醛水平差异无显著性意义(P均> 0.05)。结果表明缺血预处理能减轻肝细胞分离过程中的缺血再灌注损伤,其机制可能与其自身缺血性预适应、抗氧化、清除氧自由基的能力相关,但对供鼠肝部分切除后残肝功能的影响不明显。     

玻璃化法处理同种异体动脉的移植

背景：体外血管预处理及保存方法有多种,经过学者们长期大量的研究观察,各种方法均得到了改进,但是仍然存在弊端。因此找到一种更有效的或者几种体外血管预处理保存方法的结合,还需要大量的研究实践。 目的：拟通过比较玻璃化法和传统冷冻法保存处理的同种异体血管移植后效果,找出一种更为实用且制作简便的血管处理方法。 方法：选用健康青紫兰兔96只,手术切除双侧股动脉,根据不同体外血管预处理方法将实验分为3组,即新鲜血管自体移植组、冷冻+辐照预处理血管同种异体移植组及玻璃化+辐照预处理血管同种异体移植组。血管移植后1,2,8,12周,每组取6只动物进行腹主动脉数字减影血管造影、扫描电子显微镜、组织病理学观察,观察移植血管通畅率、动脉瘤形成情况及组织形态学变化。 结果与结论：移植后12周,新鲜血管自体移植组血管的累计通畅率显著高于冷冻+辐照预处理组(P < 0.05),玻璃化+辐照预处理组与其他两组比较,差异无显著性意义(P > 0.05)。组织病理学检查显示,玻璃化+辐照处理的同种异体血管内膜及中层平滑肌增生较冷冻+辐照预处理组轻,管腔狭窄不明显,炎症反应较轻。经玻璃化法+辐照保存的同种异体血管制作程序简单,移植血管通畅率高,组织反应轻,是一种比较理想的同种异体血管体外处理方法。     

肾上腺髓质素基因注入大鼠骨骼肌对肢体缺血再灌注肝脏的影响

背景：以往研究直接采用肾上腺髓质素药物来研究多种器官缺血再灌注损伤后肾上腺髓质素的保护作用,但转肾上腺髓质素基因对肢体缺血再灌注后引起的肝脏损伤是否有保护作用却少见报道。 目的：探讨大鼠肾上腺髓质素真核表达载体对大鼠肢体缺血再灌注后肝脏的作用。 方法：成年雄性SD大鼠32只,随机分成4组,假手术组、缺血再灌注模型组、转肾上腺髓质素基因组和空载体组,每组8只,除假手术组其余各组大鼠制成缺血再灌注模型。转肾上腺髓质素基因组和空载体组采用直接注射法于腓肠肌分别注射肾上腺髓质素真核表达载体和肾上腺髓质素(700 μg/kg)盐水溶液1 mL;假手术组和缺血再灌注模型组注射生理盐水1 mL心脏采血测定血浆谷丙转氨酶、谷草转氨酶和乳酸脱氢酶的活性,测定肝组织总超氧化物歧化酶、丙二醛的含量;免疫组化检测术侧腓肠肌中肾上腺髓质素的表达情况;显微镜观察肝组织的形态学变化。 结果与结论：与假手术组比较：缺血再灌注模型组血浆谷丙转氨酶、谷草转氨酶、乳酸脱氢酶水平及肝组织丙二醛含量均明显升高(P < 0.01),肝组织超氧化物歧化酶活性明显降低(P < 0.01),显微镜下可见肝细胞水肿、结构紊乱等组织损伤明显;与缺血再灌注模型组比较：转肾上腺髓质素基因组血浆谷丙转氨酶、谷草转氨酶、乳酸脱氢酶及肝组织丙二醛含量明显降低(P < 0.01),肝组织超氧化物歧化酶活性明显升高(P < 0.01),显微镜下可见肝细胞水肿等组织损伤有所改善。免疫组化结果显示转肾上腺髓质素基因组腓肠肌中肾上腺髓质素表达上调。结果表明,转肾上腺髓质素基因对大鼠肢体缺血再灌注后肝脏具有保护作用,其机制可能与提高抗氧化能力、降低肝酶渗出相关。     

诱导血红素加氧酶1高表达对减体积肝移植大鼠生存的影响

背景：血红素加氧酶1在防止和减轻缺血再灌注损伤方面扮演着重要角色，成为目前肝移植领域的研究热点。 目的：构建重组腺病毒Ad5-血红素加氧酶1，对肝移植供体进行预处理来诱导供肝血红素加氧酶1高表达，进一步观察其对减体积肝移植大鼠生存时间及肝功能的影响。 设计、时间及地点：随机对照动物实验，于2003-09/2005-03在重庆医科大学附属儿童医院儿科研究所完成。 材料：选用SD大鼠74只，制备原位减体积肝移植模型。 方法：利用分子生物学方法构建携带有血红素加氧酶1基因的重组腺病毒Ad5-血红素加氧酶1，对供体进行预处理。将74只大鼠按不同处理方案分组如下：生理盐水组（n=12）、诱导剂原卟啉钴组（n=13）、Ad5-血红素加氧酶1组（n=13）、Ad5-绿色荧光蛋白组（n=12）、Ad5-血红素加氧酶1+抑制剂原卟啉锌组（n=12）分别于取肝前注射生理盐水、诱导剂原卟啉钴、Ad5-血红素加氧酶1、Ad5-绿色荧光蛋白及Ad5-血红素加氧酶1+抑制剂原卟啉锌，取肝后HTK液4 ℃保存24 h后植入。对照组（n=12）取肝前48 h静脉注射生理盐水，取肝后马上植入。 主要观察指标：各组肝移植大鼠的存活率，肝功能指标测定，彩色多普勒超声监测移植后2 h门静脉血流变化，应用常规苏木精-伊红染色法观察移植肝组织病理学变化，肝组织血红素加氧酶1活性测定，应用免疫组织化学染色法检测血红素加氧酶1、肿瘤坏死因子α，bcl-2，bax在移植肝组织中的表达，从分子水平检测血红素加氧酶1、肿瘤坏死因子α，bcl-2，bax mRNA的表达变化。 结果：Ad5-血红素加氧酶1组大鼠肝移植后1，7，21 d的生存率显著高于生理盐水组 （P < 0.05）。与生理盐水组、Ad5-绿色荧光蛋白组及Ad5-血红素加氧酶1+抑制剂原卟啉锌组比较，Ad5-血红素加氧酶1组移植肝的谷丙转氨酶活性显著降低（P < 0.05），移植后2 h内门静脉血流量明显增加（P < 0.05），血红素加氧酶1活性显著升高（P < 0.05），反转录-聚合酶链反应和免疫组化检测中血红素加氧酶1、Bcl-2表达水平增高 （P < 0.05），Bax、肿瘤坏死因子α表达水平降低（P < 0.05）。 结论：构建重组腺病毒Ad5-血红素加氧酶1可以诱导供肝血红素加氧酶1高表达，显著增加大鼠减体积肝移植后2 h内门静脉血流，促进移植肝功能的恢复，从而延长肝移植后的生存时间。     

血红素氧合酶-1/一氧化碳系统在新生大鼠缺氧缺血性脑损伤中的作用

目的 观察新生大鼠缺氧缺血性脑损伤(HIBD)后血红素氧合酶-1/一氧化碳系统(HO-1/CO)变化,探讨其在HIBD中的作用.方法 7日龄Wistar新生大鼠按随机数字表法分为假手术组(Sham组)、缺氧缺血组(HIBD组)及HO抑制剂锌原卟啉组(Znpp组),每组6只.采用实时荧光定量PCR法、硫代巴比妥酸法(TBA法)、流式细胞术(FCM)和双波长定量测定法分别检测脑组织中HO-1 mRNA的表达、丙二醛(MDA)含量、脑组织细胞凋亡率及血中CO浓度.结果 与Sham组比较,HIBD组、Znpp组HO-1 mRNA表达均增强(分别为0.166±0.042、2.289±0.333、1.839±0.322),CO浓度升高[分别为(0.460±0.009)%、(1.026±0.145)%、(0.735±0.079)%],差异有统计学意义(P＜0.05);但与HIBD组比较,Znpp组HO-1 mRNA表达明显减少,CO浓度降低,差异有统计学意义(P＜0.05).与Sham组比较,HIBD组、Znpp组MDA含量及细胞凋亡率均明显升高[MDA含量分别为(1.016±0.210)nmol/mg prot、(1.945±0.312)nmol/mg prot、(3.202±0.693)nmol/mg prot,凋亡率分别为(0.108±0.009)%、(1.412±0.307)%、(2.458±0.565)%],差异有统计学意义(P＜0.05);与HIBD组比较,Znpp组MDA含量及细胞凋亡率明显增高,差异有统计学意义(P＜0.05).结论 缺氧缺血性脑损伤后HO-1/CO系统可能在脑损伤的恢复中起到一定的保护作用.     

Effect of zinc protoporphyrin on carbon monoxide/heme oxygenase-1 system in rats subjected to recurrent febrile convulsion

BACKGROUND: Studies on febrile convulsion (FC)-caused brain injury are disputed in many aspects. How FC cause nervous system injury in the developmental period and what are the characteristics of these pathological injury are unknown. The current studies have demonstrated that heme oxygenase-1 (HO-1) exerts effects on brain injury mainly by catalyzing hemoglobin to produce degradation products, and HO-1 not only has neuroprotective effects, but also has neurotoxic effects during the FC-caused brain injury. Study on the effect of zinc protoporphyrin (ZnPP) on brain injury is still in the stage of animal experiment. OBJECTIVE: To observe the effects of ZnPP on carbon monoxide (CO)/HO-1 system of rats subjected to FC, and to analyze the action pathway of ZnPP in brain protective effect. DESIGN: A randomized controlled animal experiment. SETTING: Department of Pediatrics, First Hospital Affiliated to Jiamusi University. MATERIALS: Sixty-five Wistar rats, of either gender, were involved in this study. They were randomized into normal control group( n =14, 37 ℃ water bath) and febrile treatment group (n =51, 44.5 ℃ hot water bath). Febrile treatment group was sub-divided into febrile non-convulsion group (FNC group, n =16) and FC group (n =35). FC group was further sub-divided into simple convulsion group (n =20) and ZnPP treatment group (n =15). HO-1 mRNA in situ hybridization kit was provided by Boster Bioengineering Co.,Ltd. ZnPP(dark brown powder) was the product of Jingmei Bioengineering Company. METHODS: This study was carried out in the postgraduate laboratory of Jiamusi University between January 2004 and January 2007. Rats in the febrile treatment group were placed in the 44.5 ℃ hot water bath box. If rats did not convulse in the water within 5 minutes, they were taken out, namely FNC group (n = 16), and those, which were convulsed within 5 minutes, were taken out immediately when they presented such a phenomenon, namely FC group (n =35). Convulsion induction was conducted once every other day, totally 10 times. Rats were euthanized for analysis at 24 hours after the last induction. Rats in the control group were placed in the 37 ℃ water. Rats in the ZnPP treatment group were intraperitoneally injected with ZnPP at 45 μmol/kg before FC attack. Rats in the simple convulsion group were only induced to be convulsed but not administrated. MAIN OUTCOME MEASURES: CO level in the brain tissue homogenate and plasma of rats in each group was detected with a spectrophotometer. HO-1 mRNA expression in the hippocampal CA1 region, CA3 region and dentate gyrus of rats was observed by in situ hybridization technique. RESULTS: Sixty-five Wistar rats were involved in the study. Two rats died respectively due to drowning and convulsion in the FC group. One rat died due to convulsion drowning in the ZnPP treatment group. ① Plasma CO concentration of control group and ZnPP treatment group was significantly lower than that of the FC group (P < 0.01), and was significantly higher in the ZnPP treatment group than in the FNC group (P < 0.05). ② CO level in the brain tissue homogenate was significantly lower in the control group and ZnPP treatment group than in the FC group (P < 0.01), and was very significantly higher in the ZnPP treatment group than in the control group (P < 0.01). ③ HO-1 mRNA expressions in the neuron of hippocampal CA1 region, CA3 region and dentate gyrus of the control group were the lowerest, and those in the FC group were the highest. HO-1 mRNA expression in the neuron of dentate gyrus in the FC group was significantly higher than that in the ZnPP treatment group (P < 0.01), and those in the FNC group and control group was significantly lower than that in the ZnPP treatment group (P < 0.01). CONCLUSION: FC can cause brain injury. Over-expression of HO-1 mRNA and the increase of CO are involved in the patho-physiological process of FC. ZnPP can inhibit HO-1mRNA activity and decrease CO level, which is one of pathways for protecting brain.     

Electroacupuncture Ameliorates Intestinal Barrier Destruction in Mice With Bile Duct Ligation–Induced Liver Injury by Activating the Cholinergic Anti-Inflammatory Pathway

ObjectivesElectroacupuncture (EA) at Zusanli (ST36) can attenuate inflammation in different rodent models. However, the therapeutic mechanisms underlying its action in inhibiting intestinal barrier destruction and liver injury in cholestasis mice have not been clarified. This study aimed at investigating whether EA at ST36 could activate the cholinergic anti-inflammatory pathway to inhibit intestinal barrier destruction and liver injury in cholestasis mice.Materials and MethodsMale Hmox1^floxp/floxp C57BL/6 mice were randomized and subjected to a sham or bile duct ligation (BDL) surgery. The BDL mice were randomized and treated with, or without (BDL group), sham EA at ST36 (BDL+sham-ST36) or EA at ST36 (BDL+ST36), or received α-bungarotoxin (α-BGT), a specific inhibitor of nicotinic acetylcholine receptor α7 subunit (α7nAChR), before stimulation (BDL+ST36+α-BGT). These mice, together with a group of intestine-specific heme oxygenase-1 (HO-1) knockout (KO) Villin-Cre-HO-1^?/? mice, were monitored for their body weights before and 14 days after BDL. The levels of plasma cytokines and liver injury–related alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by enzyme-linked immunoassay, and pathological changes in the intestinal mucosa and liver fibrosis as well as intestinal barrier permeability in individual mice were examined by histology and immunohistochemistry. The levels of α7nAChR, HO-1, ZO-1, Occludin, Claudin-1, and NF-κBp65 expression and NF-κBp65 phosphorylation in intestinal tissues were quantified.ResultsCompared with the sham group, BDL significantly increased the levels of plasma interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor α, ALT, and AST and caused intestinal mucosal damages, high permeability, and liver fibrosis in mice, which were remarkably mitigated, except for further increased levels of plasma IL-10 in the BDL+ST36 group of mice. Similarly, EA at ST36 significantly up-regulated α7nAChR and HO-1 expression; mitigated the BDL-decreased ZO-1, Occludin, and Claudin-1 expression; and attenuated the BDL-increased NF-κBp65 phosphorylation in intestinal tissues of mice. The therapeutic effects of EA at ST36 were significantly abrogated by pretreatment with α-BGT or HO-1 KO.ConclusionEA at ST36 inhibits the BDL-induced intestinal mucosal damage and liver fibrosis by activating the HO-1 cholinergic anti-inflammatory pathway in intestinal tissues of mice.     

大鼠近交系间原位肝移植急性排斥模型的建立与评价

背景：目前国内常用的封闭群品系大鼠(SD与Wistar大鼠)所建立肝移植急性排斥模型并不理想,易产生肝移植耐受。 目的：通过二袖套法建立稳定的DA-Lewis大鼠原位肝移植急性排斥模型。 方法：实验分为两组：同基因组：Lewis-Lewis 24例;异基因组：DA-Lewis 24例。观察移植后一般情况及移植后存活时间,两组受体分别于移植后3,5,7,10 d随机取3只处死取标本,观察肝脏组织病理变化,测定天冬氨酸转氨酶、总胆红素、细胞因子水平变化。 结果与结论：同基因组大鼠无急性排斥反应表现,中位生存时间超过100 d,肝脏组织发生轻度形态学改变。异基因组大鼠移植后黄疸明显,中位生存时间为11 d,移植后第7天肝脏组织病理表现典型急性排斥反应(Banff国际标准)。同期相比异基因组天冬氨酸转氨酶、总胆红素、细胞因子水平均高于同基因组(P < 0.001)。提示,DA-Lewis是稳定的大鼠肝移植急性排斥模型,是研究肝移植排斥反应及免疫耐受的理想动物模型。     

Changes of PACAP Immunoreactivities and Cytokine Levels After PACAP-38 Containing Intestinal Preservation and Autotransplantation

Small bowel is one of the most sensitive organs to ischemia–reperfusion injury, which is a significant problem during transplantation. Pituitary adenylate cyclase-activating polypeptide (PACAP) has cytoprotective effect in ischemic injuries of various tissues. The aim of our study was to measure changes of PACAP-38 and PACAP-27 immunoreactivities and cytokine levels in intestinal grafts stored in PACAP-38-containing preservation solution. Small bowel autotransplantation was performed on male Wistar rats. Grafts were stored in University of Wisconsin (UW) solution at 4 °C for 1 h (group (G)I), for 3 h (GII), and for 6 h (GIII) and in PACAP-38-containing UW solution for 1 h (GIV), for 3 h (GV), and for 6 h (GVI). After preservation, performing vessel anastomosis reperfusion began, which lasted 3 h in each group. Tissue biopsies were collected after laparotomy (control) and at the end of the reperfusion periods. Intestinal PACAP-38 and PACAP-27 immunoreactivities were measured by radioimmunoassay. To measure cytokines from tissue homogenates, we used rat cytokine array and Luminex Multiplex Immunoassay. Levels of PACAP-38 and PACAP-27 immunoreactivity decreased after 1 and 3 h preservation compared to control levels. This decrease was significant following 6 h cold storage (p?<?0.05). Values remained significantly higher in grafts stored in PACAP-38-containing UW. Cytokine array revealed that expression of the soluble intercellular adhesion molecule-1 (CD54) and L-selectin (CD62L/LECAM-1) was increased in GIII. Both 6 h cold storage in PACAP-38-containing UW solution and 3 h reperfusion caused strong reduction in these cytokines activation in GVI. RANTES (CCL5) levels were increased in all groups. Strong activation of the tissue inhibitor of metalloproteinase-1 was in GIII. However, PACAP-38-containing cold storage could decrease its activation in GVI. Furthermore, strong activation of the tissue inhibitor of metalloproteinase-1 was detected in 6 h preserved grafts without PACAP-38 (GIII). PACAP-38-containing cold storage could decrease its activation in GVI. Our present study showed that PACAP-38 and PACAP-27 immunoreactivities decreased in a time-dependent manner during intestinal cold preservation, which could be ameliorated by administration of exogenous PACAP-38 to the preservation solution. Moreover, PACAP-38 could attenuate tissue cold ischemic injury-induced changes in cytokine expression.