大鼠肝动脉永久性或间歇阻断后侧枝循环的观察

永久性肝动脉结扎（ＰＨＡＬ）后侧枝循环的建立．是ＰＨＡＬ治疗肝癌效果不持久的原因之一．如何防止或减少侧枝循环的形成．是肝癌缺血治疗的重要环节之一。通过大鼠选择性腹腔动脉造影（ＳＣＡ）的方法．观察ＰＨＡＬ和肝动脉反复暂时阻断（ＲＴＢＨＡ）后侧枝形成的情况。ＰＨＡＬ后２ｗｋＳＣＡ即可见侧枝循环．随着时间延长．侧枝愈加明显丰富，其主要来源是肠系膜上动脉、腹腔动脉和大网膜。ＲＴＢＨＡ后２ｗｋ，ＳＣＡ未显示侧枝形成。实验结果表明肝动脉的间歇阻断可以防止侧枝循环的建立。这为ＲＴＢＨＡ的临床运用提供了实验依据。     

ATP—MgCL2对犬肝脏常温下缺血后的保护作用

常温下阻断肝脏血流６０分钟后立即恢复肝脏血流，治疗组犬静脉内给予ＡＴＰ－ＭｇＣｌ２，对照组犬给予生理盐水。恢复肝血流后一小时，治疗组ＳＧＰＴ较对照组低（Ｐ＜０．０５）；光镜下治疗组犬肝细胞浊肿、糖原颗粒较多，对照组犬肝细胞脂肪变性及片状坏死、糖元颗粒消失；电镜下治疗组犬肝线粒体形态规则、内膜嵴可见，对照组犬肝线粒体肿胀、嵴断裂、消失、基质空泡状；犬的存活率治疗组显著高于对照组（Ｐ＜０．０５）。本结果证实，ＡＴＰ－Ｎｇｃｌ２能改善缺血后的肝细胞的功能和结构及动物的存活率。     

肝切除时门静脉血部分动脉化的研究

目的 研究犬门静脉血部分动脉化的肝保护作用。方法 建立大保留肝（占全肝６０％）暂时性血流阻断、肝固有动脉切断并切除未阻断肝的急性肝衰模型（对照组）,并行肝总动脉与胃十二指肠静脉吻合（Ａ－Ｐ组）,观察生存率并定时测定丙氨酸转氨酶（ＡＬＴ）、动脉血酮体比（ＡＫＢＲ）及肝动脉脉、门静脉血气分析。结果 对照组７天生存率为３７．５％,Ａ－Ｐ组均较差异有非常显著性（Ｐ〈０．０１）,门静脉和肝静脉血氧分压均较术     

反复暂时性去肝动脉化治疗晚期肝癌

作者与瑞典Ｌｕｎｄ大学医学院合作，前瞻性对比研究反复暂时性去肝动脉化与肝动脉结扎对晚期肝癌的疗效。作者将１９９４年２月至１９９５年４月相继入院的４０例经手术和病理确诊的不可切除的肝细胞型肝癌患者，随机分为两组，每组２０例，分别采用肝动脉结扎（ＨＡＬ）或反复暂时性去肝动脉化术（ＲＴＤ）治疗，对术后反应、生存期、术后肝功能、甲胎蛋白浓度和影象学检查进行比较。结果表明ＲＴＤ组优于ＨＡＬ组。ＲＴＤ组与ＨＡ     

盐酸氟桂利嗪对再灌注损伤时鼠肝细胞线粒体的作用

目的 盐酸氟桂利嗪对大鼠肝脏缺血／再灌注香肝细胞线粒体的影响。方法：将Ｗｉｓｔａｒ大鼠随机分为三组,每组１２只。Ａ组为对照组,Ｂ组缺血／再灌注组组,Ｃ组为盐酸氟桂利嗪组。其中Ｂ,Ｃ两组均阻断肝门造成肝脏完全缺血３０分钟后再灌注９０分钟。测定各组动物血清中ＡＬＴ,ＬＤＨ含量,肝细胞线粒体脂质过氧化物（ＬＰＯ）含量、行超微结构的观察。结果 与Ａ组比较,Ｂ组血清ＡＬＴ,ＬＤＨ活性显著加（Ａ,Ｂ二组,ＡＬ     

深低温循环中断对血浆内源性洋地黄物质含量变化及对心肌超

实验犬１５只，随机分为三组，观察深低温循环中断不同时限（Ａ组３０分钟，Ｂ组６０分钟，Ｃ组８０分钟）及复温复跳后血浆内源性洋地黄物质变化和心超微结构的改变，结果表明，全组犬血浆ＥＤＬＳ在麻醉后即有所降低，转流前降至最低水平。Ｃ组在循环停止末期血浆ＥＤＬＳ显著增高，复温复跳后进一步升高，与ＰＨ、ＰＯ２有显著的负相关。Ｃ组犬心肌超微结构明显损害。实验提示，心肌能耐受急性缺血缺氧仍以６０分钟为安全限度。Ｅ     

深低温循环中断对血浆内源性洋地黄物质含量变化及对心肌超微结构的影响

实验犬１５只，随机分为三组。观察深低温循环中断不同时限（Ａ组３０分钟，Ｂ组６０分钟，Ｃ组８０分钟）及复温复跳后血浆内源性洋地黄物质（ＥＤＬＳ）变化和心肌超微结构的改变，结果表明，全组犬血浆ＥＤＬＳ在麻醉后即有所降低，转流前降至最低水平。Ｃ组在循环停止末期血浆ＥＤＬＳ显著增高，复温复跳后进一步升高，与ｐＨ、ＰＯ２显著的负相关。Ｃ组犬心肌超微结构明显损害。实验提示，心肌能耐受急性缺血缺氧仍以６０分钟为安全限度。ＥＤＬＳ急剧增高是重要脏器细胞遭受严重缺血缺氧损害的标志。     

人肝细胞系生物功能筛选测定

生物人工肝利用原代肝细胞或人肝细胞系提供肝功能支持［１～４］。我国是世界上最早建立人肝细胞系的国家之一［５］。本研究收集国内的９株人肝细胞系,从中筛选出分化程度高者,测定其生物功能,以备作为人工肝的生物材料。材料与方法１．材料９株人肝细胞系由中国科学院上海细胞生物学研究所提供,编号为细胞系１～９（ＣＬ１～９）。原代肝细胞取自５月胎龄水囊引产的人胚胎肝组织。Ｌ－１５培养基为Ｇｉｂｃｏ产品。Ｌ－谷氨酰胺为Ｓｉｇｍａ产品。ＦＡＣＳ　４４０流式细胞仪为美国Ｂｅｃｔｏｎ　Ｄｉｃｋｉｎｓｏｎ公司产品。２．方…     

促肝细胞生长素对肝硬变大鼠部分肝切除后肝再生影响的实验研究

为观察促肝细胞生长素（ｐＨＧＦ）对肝硬变大鼠部分肝切除后肝再生的影响，应用ＩＢＡＳⅡ全自动图像分析仪检测肝硬变大鼠术后肝细胞ＤＮＡ倍体率及肝脏组化染色后各组酶灰度值（ＯＰＴＤＭ）的变化，并通过检测血清谷丙转氨酶（ＳＧＰＴ）及吲哚青绿１５分钟滞留率（ＩＣＧ１５）以了解残肝功能。结果：实验组（Ａ组）３～４倍体肝细胞明显减少，２倍体及多倍体肝细胞明显增多；术后第１天各组酶灰度值无显著差异，第２、５天琥珀酸脱氢酶（ＳＤＨ）和乳酸脱氢酶（ＬＤＨ）明显增高，而碱性磷酸酶（ＡｋＰ）和酸性磷酸酶（ＡｃＰ）则明显减少；Ａ组术后第５天ＳＧＰＴ和ＩＣＧ１５明显降低。本实验结果提示：ｐＨＧＦ不仅能促使肝硬变大鼠肝切除后残肝的再生，而且能够促进再生肝细胞功能成熟及残肝功能的恢复     

混合型生物人工肝治疗急性肝衰犬的研究

我们在成功构建生物反应器 ,完成体外灌流实验基础上[1 ] ,建立混合型生物人工肝支持系统 (ＨＢＬＳ) ,对急性肝衰犬模型进行实验性治疗 ,以测试其效能。一、材料与方法1.实验动物 :杂种犬 7条 ,体重 15～18ｋｇ ,行门腔静脉吻合 ,肝蒂预置套带后关腹 ;48ｈ后再阻断肝蒂 1ｈ做成肝衰模型。 4条用ＨＢＬＳ治疗 ,另 3条空舱转流作为对照。2 .生物反应器的构成 :宁波亚泰Ⅱ级血浆分离器 (分子量截留为 75 0 0 0Ｄａｌｔｏｎｓ ,双醋酸纤维 ) ,纤维外间隙容积为 15 0ｍｌ ,内置 1.2× 10 9的数量级微载体培养Ｌ 0 2人肝细胞[2 ] 。3 .ＨＢＬＳ构…     

肝血流阻断方法在肝硬化患者肝肿瘤手术中的选择

目的:评价肝硬化患者行肝脏肿瘤切除术时肝血流阻断方法的选用。方法:将我院2007年6月—2012年12月63例需行肝切除的肝肿瘤伴肝硬化患者随机分为3组。A组22例,为间歇血流阻断组,采用阻断肝血流15 min,复流5 min;B组20例,为半肝血流阻断组,行同侧半肝血流持续性阻断,健侧血流正常;IP组21例,采用入肝血流阻断前先阻断5 min、复流5 min的预处理方法。比较3组术中阻断时间、肝断面单位面积出血量、3组及3组中阻断时间<30 min者术后1、3、7、14 d的ALT、AST及TB变化情况。结果:3组肝断面单位面积出血量差异有统计学意义(P<0.05)。术后B组缺血再灌注损伤最轻,IP组最重,差异有统计学意义(P<0.05)。阻断时间<30min者术后肝功比较,IP组较A组缺血再灌注损伤明显降低(P<0.05)。结论:肝血流阻断前预处理可减少肝切除术中的失血量。术前预计切除时间<30 min的患者可采用预处理减少出血量及再灌注损伤。     

Bile duct exclusion from selective vascular inflow occlusion in rat liver: role of ischemic preconditioning and N-acetylcysteine on hepatic reperfusion injury

AIM: To study the effects of N-acetylcysteine and ischemic preconditioning on the portal triad clamping compared to arterial and portal clamping alone. METHODS: Eighty EPM 1-Wistar rats were randomized into two groups, depending on inclusion (Group 1) or not (Group 2) of the bile duct in the hepatic vascular pedicle occlusion. Each group was divided into four subgroups as follows. IR 1: 20 minutes after celiotomy, the pedicle containing vascular elements and bile duct to the left lateral and median liver lobes was occluded for 40 minutes, followed by 30 minutes of reperfusion. IPC 1: after 10 minutes of ischemia and 10 minutes of reperfusion, the ischemic preconditioning period, the rats were submitted to the same procedure described for IR 1 Group. NAC 1: the rats received N-acetylcysteine (150 mg/kg) 15 minutes before 40 minutes of ischemia and 5 minutes before 30 minutes of reperfusion. SHAM 1: The hepatic pedicle for the lateral and median liver lobes was dissected after 20 minutes, the bile duct alone was clamped for 40 minutes, and released for an additional 30 minutes. In the IR 2, IPC 2, and NAC 2 groups, ischemia was achieved with an exclusive vascular occlusion. SHAM 2: dissection and observation for 90 minutes. The blood was sampled for liver enzyme levels. Statistical analysis was done (P 相似文献   

奥曲肽预处理对围术期肝脏缺血-再灌注损伤保护作用的临床研究

目的探讨奥曲肽预处理对围术期肝脏缺血-再灌注损伤保护作用及可能的机制。方法选择88例肝脏手术患者,随机分为研究组45例,对照组43例,术中行肝门阻断。研究组患者于手术前1h给予奥曲肽0.2mg皮下注射,对照组于手术前1h给予生理盐水1ml皮下注射。观察术前(T0)、术后6h(T1)、24h(T2)及7d(T3)时血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH),肿瘤坏死因子α(TNF-α)及白细胞介素1β(IL-1β)的变化,手术部位切除后取标本行超氧化物歧化酶(SOD)、髓过氧化物酶(MPO)的检测。采用TUNEL法检测肝组织中的凋亡细胞数。结果 T0时两组之间ALT、AST、LDH及TNF-α、IL-1β差异无统计学意义。T1时两组ALT、AST、LDH及TNF-α、IL-1β明显高于T0时(P<0.05),对照组又明显高于研究组(P<0.05)。T2时两组ALT、AST、LDH开始下降,但仍然高于T0时(P<0.05),T3时恢复正常范围,而T2、T3时TNF-α、IL-1β降至正常范围。研究组肝组织MPO明显低于对照组(P<0.05),而SOD明显高于对照组(P<0.05)。肝组织中的凋亡细胞数对照组为(67.79±5.25)明显高于研究组(44.32±5.16)(P<0.01)。结论奥曲肽对围术期肝脏缺血-再灌注损伤有保护作用,其可能的机制为稳定细胞膜、抑制炎性反应及细胞凋亡。     

Kallistatin对大鼠肝缺血再灌注损伤的保护及机制研究

目的:探讨Kallistatin对大鼠肝缺血再灌注后肝损伤的保护作用及机制。方法:建立大鼠肝脏缺血再灌注模型,随机将Wistar大鼠分成假手术组(S组)、缺血再灌注组(I/R组)、空壳腺病毒组(O组)和Kallistatin组(K组)。测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平评价肝功能。HE染色观察肝组织病理学改变。TUNEL法观察肝细胞凋亡情况。Westen-blot法检测肝组织中Caspase-3、Bcl-2、Bax的表达。结果:与S组比较,I/R组AST、ALT水平明显升高(P<0.05),病理损伤严重,Cas-pase-3和Bax表达也相应增加(P<0.05),而Bcl-2表达减少(P<0.05)。与I/R组比较,K组各项指标均明显改善(均为P<0.05)。O组与I/R组间无明显差别(P>0.05)。结论:Kallistatin对大鼠肝脏缺血再灌注后具有明显的保护作用,其机制可能通过促进Bcl-2表达、抑制Bax表达实现。     

Hepatic arterial buffer response fails to restore hepatic oxygenation after temporary liver dearterialization in canines

BACKGROUND: Hepatic artery thrombosis is a rare but extremely troublesome condition after liver transplantation. Recently, urgent arterial revascularization has been used as rescue therapy, leading to improved graft and patient survivals. Hepatic artery ligation produces a progressive reduction in portal vein blood flow. Theoretically, a hyperemic response may be expected following hepatic artery reperfusion (hepatic artery buffer response, HABR). In this study, we tested the hypothesis that HABR can maintain adequate liver oxygenation after temporary liver dearterialization. METHODS: Seven dogs (19.7 +/- 1.2 kg) subjected to 60 minutes of hepatic artery occlusion were observed for 120 minutes thereafter. Systemic hemodynamics was evaluated through Swan-Ganz and arterial catheters, and splanchnic perfusion by portal vein and hepatic artery blood flows (PVBF and HABF) via an ultrasonic flowprobe. Liver enzymes (ALT and LDH) and systemic and hepatic oxygen delivery (DO2hepat) were calculated using standard formulae. RESULTS: Hepatic artery occlusion induced a progressive reduction in PVBF and DO2hepat. A complete restoration of HABF after hepatic artery declamping was observed; however, the DO2hepat (33.3 +/- 5.9 to 16.5 +/- 5.9 mL/min) did not return to the baseline levels. CONCLUSION: Temporary hepatic artery occlusion induced a progressive decrease in portal vein blood flow during ischemia, an effect that continued during the reperfusion period. The hepatic artery blood flow was promptly restored after declamping. However, HABR was not able to restore hepatic oxygen delivery to baseline levels during the reperfusion period.     

瑞芬太尼对肝硬化大鼠肝脏缺血再灌注损伤的影响

目的 探讨瑞芬太尼对肝硬化大鼠肝脏缺血再灌注损伤的影响.方法 成年健康雄性SD大鼠30只,体重260～300 g,采用随机数字表法,将其随机分为3组(n=10):肝硬化组(C组)、肝硬化+肝缺血再灌注组(I/R组)和瑞芬太尼组(R组).C组、I/R组和R组采用四因素综合法制备大鼠肝硬化模型,I/R组和R组在肝硬化模型制备成功后1周制备大鼠70％肝脏缺血再灌注模型,R组于缺血前10 min开始静脉输注瑞芬太尼1μg·kg-1·min-至再灌注结束.于再灌注4h时取静脉血样和肝组织,测定血清ALT和AST活性、肝细胞Bcl-2和Bax表达及肝细胞凋亡情况,计算细胞凋亡指数,光镜下观察肝组织病理学结果.结果 与C组比较,I/R组血清ALT和AST的活性升高,肝细胞Bcl-2表达下调,Bax表达上调,细胞凋亡指数升高(P＜0.05);与I/R组比较,R组血清ALT和AST的活性降低,肝细胞Bcl-2表达上调,Bax表达下调,细胞凋亡指数降低(P＜0.05).R组肝组织病理学损伤轻于I/R组.结论 瑞芬太尼可减轻肝硬化大鼠肝脏缺血再灌注损伤,其机制与平衡肝细胞Bcl-2与Bax表达而抑制肝细胞凋亡有关.     

大黄素对大鼠肝脏缺血再灌注损伤的预防作用

目的:探讨大黄素对肝脏缺血-再灌注损伤的保护作用及其机制.方法:将45只成年雄性SD大鼠随机分成三组:假手术对照组(A组)15只,肝缺血30 min、再灌注90min组(B组)15只,术前5 d给予大黄素灌胃60mg/(kg·d),5次 肝缺血30 min、再灌注90min(C组)15只;观察血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、透明质酸酶(HA)、肝组织丙二醛(MDA)含量的变化及肝脏组织病理学改变情况,同时测定各组大鼠胆汁流量情况.结果:C组与B组相比,MDA含量显著降低(P＜0.05),HA浓度明显降低(P＜0.01),胆汁流量显著增加(P＜0.01),同时C组的肝细胞功能明显改善,且肝脏超微结构改变也较轻.结论:大黄素对肝脏缺血-再灌注损伤具有保护作用,该作用与减轻肝脏缺血再灌注后脂质过氧化程度和肝窦内皮细胞损伤有关.     

黄芪注射液对肝门阻断后肝功能损害的治疗作用

目的探讨黄芪注射液对肝门阻断后肝脏的治疗保护作用。方法88例肝门阻断肝脏手术的病人随机分为黄芪注射液治疗组（Ⅰ组）和非黄芪注射液治疗组（Ⅱ组）,术前和术后3d、7d分别检测直接胆红素（DBil）、碱性磷酸酶（ALP）、天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、7谷氨酰转移酶（GGT）、白蛋白（Alb）,两组资料进行对比分析。结果与Ⅱ组比较,Ⅰ组术后3d、7dAST、ALT明显恢复,差异有统计学意义。结论黄芪注射液对肝门阻断后的肝脏缺血缺氧及再灌注损伤有一定的保护作用。     

c-Jun N-terminal kinase mediates hepatic injury after rat liver transplantation

BACKGROUND: Orthotopic liver transplantation (OLT) requires cold ischemic storage followed by warm reperfusion. Although c-Jun N-terminal kinase (JNK) is rapidly activated after OLT, the functional consequences of JNK activation are unknown. The aim of this study was to address the role of JNK after OLT using the selective JNK inhibitor CC-401. METHODS: Donors, recipients, or stored liver explants were treated with vehicle or JNK inhibitor before OLT by an arterialized two-cuff method with 40 hours of cold storage. Recipients were assessed for 30-day survival, and graft injury was assessed over time by hepatic histology, serum transaminases, caspase 3 activation, cytosolic cytochrome c, and lipid peroxidation. RESULTS: Survival after OLT increased after donor plus storage and storage only treatment with JNK inhibitor (P<0.05). Treatment of recipient only did not improve survival. Increased survival correlated with improved hepatic histology and serum aspartate aminotransferase levels. JNK inhibition significantly decreased nonparenchymal cell killing at 60 minutes after reperfusion (P<0.05) and pericentral necrosis at 8 hours after reperfusion (P<0.01). JNK inhibition decreased cytochrome c release, caspase 3 activation (P<0.05), and lipid peroxidation (P<0.05). JNK inhibition also transiently blocked phosphorylation of c-Jun at 60 minutes after reperfusion (P<0.05) without affecting other MAPK signaling, including p-38 and Erk activation. CONCLUSIONS: JNK inhibition decreases hepatic necrosis and apoptosis after OLT, suggesting that JNK activation promotes cell death by both pathways. Inhibition of JNK may be a new therapeutic strategy to prevent liver injury after transplantation.     

Changes in serotonin and histamine levels in blood and liver after acute hepatic ischemia

To clarify the changes of vasoactive amines associated with acute hepatic failure, ammonia, tryptophan, serotonin (5-HT) and histamine in the blood and liver were studied in dogs (n = 22) of each three group of acute hepatic ischemia; occlusion of hepatic artery (controls), occlusion of hepatic artery and portal vein (THI), and portocaval shunt with THI (PCS + THI). These biochemical changes were studied in each group at six time intervals: Preocclusion, 15 and 30 minutes postocclusion, and 30, 60 and 120 minutes after release of occlusion. A rapid rise of blood ammonia levels was observed in groups of THI and PCS + THI after occlusion. Blood 5-HT increased in postocclusion of both controls and THI. However, a decrease of 5-HT was observed in PCS + THI. Hepatic 5-HT also increased after occlusion in THI and PCS + THI as compared with a decrease in controls. Plasma histamine rose significantly in all groups after the occlusion. These data demonstrated that the changes of vasoactive amines in hepatic ischemia and/or splanchnic pooling appeared to affect microcirculation of the liver and play a role of pathogenesis of hepatic failure after hepatic ischemia.