去铁敏预处理对移植供肝保护作用的研究

目的：探索去铁敏对移植供肝的保护作用及可能机制。方法：SD大鼠,随机分为假手术组（S组）、自体肝移植组（AT组）、自体肝移去铁敏预处理组（DP组）与自体肝移植去铁敏灌肝组（DI组）。检测各组术后24 h血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）和碱性磷酸酶（ALP）,观察肝组织形态学改变并检测肝组织中低氧诱导因子-1（HIF-1α）及丙二醛（MDA）的含量。结果：与S组相比,AT组、DP组和DI组术后血清ALT,AST和ALP明显升高;但DP组、DI组较AT组血清ALT,AST和ALP明显降低,肝形态异常变化减轻,肝组织中HIF-1α蛋白含量升高,MDA含量下降(P﹤0.05)。结论：在肝移植术前行去铁敏预处理或术中行去铁敏溶液灌肝,对供肝有保护作用,且前者优于后者。其机制可能与提高组织HIF-1α含量及减轻脂质过氧化有关。     

用于再灌注损伤研究的两种肝移植模型的筛选

目的:筛选一种用于研究肝移植过程中缺血再灌注损伤的大鼠肝移植模型.方法:将SD大鼠随机分为假手术组(S组)、原位肝移植组(OLT组)与自体肝移植组(AT组).比较各组大鼠手术时间、存活率及术后24 h血清中总胆红素(TB)、谷丙转氨酶(ALT)、谷草转氨酶(AST)及血清肿瘤坏死因子-α(TNF-α)含量,观察肝组织形态学改变.结果:与S组相比OLT组、AT组术后血清TB、ALT、AST升高明显,AT组较OLT组术后存活率明显提高,血清TB、ALT、AST及TNF-α含量明显降低,肝形态异常变化减轻(P<0.05).结论:自体肝移植模型手术简单、成功率高、可比性强,且排除了免疫损伤的影响,更好地反映了肝脏缺血再灌注损伤的病理生理过程,比较适宜作为肝移植缺血再灌注损伤研究的动物模型.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧与去铁敏对移植供肝作用的对比性研究

Objective To compare the protecting effect of hypoxia preconditioning(HP) and deferoxam-ine preconditioning (DP)on transplanted livers and to search a protecting method which is suitable for clini-cal practice.Methods SD rats are randomly divided into four groups:sham operation group (S),analogic transplantation group (AT),deferoxamine preconditioning group (DP) and hypoxia preconditioning group (HP).Twenty-four hours after operation,several factors of each group were measured respectively,inclu-ding deterioration of histomorphology,hepatic zymogram in the serum,hypoxia inducing factor-1 (HIF-1α)and malonaldehyde (MDA) in hepatic tissue.Results The levels of alanine aminotransferase (ALT),glu-tamic oxalacetic transaminase (AST) and alkaline phosphatase (ALP) in AT group,DP group and HP group were higher than S group's.Levels of ALT,AST,ALP in AT group were dramatically high,hepatic tissue suffers from edema,degeneration even necrosis.By contrast,the pathological change is milder in DP group and HP group.HIF-1α in hepatic tissue of AT group was lower than that in DP group and HP group,MDA was higher than two latters (P < 0.05).Conclusion DP can protect transplanted liver as same as HP,however,DP is more suitable for clinical practice.     

低氧预适应诱导肝脏HIF-1α高表达对自体原位肝移植大鼠JNK信号转导通路的影响

目的 检测低氧诱导因子α(HIF-1α)在自体原位肝移植大鼠肝脏的表达情况,探讨HIF-1α在JNK信号转导通路中的作用机制.方法 采用单纯随机抽样法将96只SD大鼠分为正常对照组、自体肝移植组、低氧预适应+自体肝移植组,每组32只.正常对照组:仅开腹分离血管,不作其他处理;自体肝移植组:采用经门静脉灌注法建立稳定的70%自体原位肝移植模型;低氧预适应+自体肝移植组:术前用8%氮氧混合气体低氧预处理90min,其他处理同自体肝移植组.分别于术后1、2、12、24h检测大鼠血清ALT、AST水平,免疫组织化学染色法测定大鼠肝脏组织HIF-1α的表达,RT-PCR检测c-Jun mRNA表达水平,Western blot检测Cleaved Caspase-3蛋白表达水平.多组比较采用方差分析,两组比较采用t检验.结果 术后1、2、12、24 h,低氧预适应+自体肝移植组大鼠血清ALT和AST水平显著低于自体肝移植组,但高于正常对照组,差异有统计学意义(F=2631.371、1177.642、810.383、682.848,743.618、1095.522、375.995、580.613,P＜0.05).自体肝移植组大鼠术后各时相点HIF-1α蛋白的表达水平明显低于低氧预适应+自体肝移植组,但均高于正常对照组,差异有统计学意义(F=191.737,284.482,459.419,213.782,P＜0.05).低氧预适应+自体肝移植组大鼠术后1、2、12 h c-Jun mRNA表达水平显著低于自体肝移植组,但均高于正常对照组,差异有统计学意义(F=66.211,53.169,9.645,P＜0.05),术后24 h c-Jun mRNA表达水平3组间比较,差异无统计学意义(F=1.100,P＞0.05).低氧预适应+自体肝移植组大鼠术后各时相点Cleaved Caspase-3蛋白表达水平显著低于自体肝移植组,但均高于正常对照组,差异有统计学意义(F=23.133,31.158,14.347,29.043,P＜0.05).结论 肝移植大鼠低氧预适应后HIF-1α的高表达可能通过抑制JNK信号通路的持续激活,下调Cleaved Caspase-3蛋白的表达,从而抑制肝细胞凋亡,减轻肝脏缺血再灌注损伤.     

低氧预适应对自体原位肝移植大鼠血清内毒素的影响

目的 探讨低氧预适应(HP)对大鼠自体肝移植后血清内毒素的影响.方法 采用改良的自体肝移植模型进行研究.将SD大鼠随机分为正常对照组(NC)、自体肝移植(AT)和低氧预适应+AT(HP)3组,每组24只.HP组术前用8%氮氧混合气体处理90 min.各组分别于术后1,6,12 h处死大鼠取肠标本,抽血检测ALT和AST水平;在光镜及透射电镜下观察各组小肠黏膜损伤程度及线粒体超微结构改变;同时应用ELISA试剂盒测定大鼠血清内毒素水平的变化.结果 HP组血清ALT,AST及内毒素水平均显著低于AT组,但高于NC组,差异均有统计学意义(P＜0.05);光镜下 HP组小肠黏膜损伤程度明显低于AT组;透射电镜下HP组线粒体损伤较AT组明显减轻.结论 HP可减轻肝IRI对肠道黏膜的损伤,减少了肠道内毒素移位而降低肝移植大鼠血清内毒素水平,从而减轻内毒素对移植肝功能的影响,其机制可能与减轻线粒体结构损伤,减轻缺血缺氧后线粒体功能障碍,改善细胞的供能,提高细胞的抗缺氧能力有关.     

低氧预适应对自体肝移植大鼠Bcl-2蛋白表达的影响及其意义

目的 探讨低氧预适应(HP)对大鼠自体肝移植后Bcl-2蛋白表达的影响及其意义.方法 采用改良的自体肝移植模型研究缺血再灌注损伤(IRI),将SD大鼠随机分为正常对照(NC)、自体移植(AT)和AT+低氧预适应(HP)3组,每组24只.HP组术前用8%氮氧混合气体处理90 min,分别于术后1,6,24 h处死大鼠取肝脏标本,抽血检测肝功能,免疫组化方法测定大鼠肝组织Bcl-2蛋白的表达,并在光镜和透射电镜下观察各组肝细胞形态学改变、线粒体损伤程度等.结果 HP组血清ALT和AST水平显著低于AT组,差异有统计学意义 (P＜0.05);HP组各时段的肝功能明显优于AT组;HP组各时段Bcl-2蛋白明显高于AT组,差异有统计学意义(P＜0.05).光镜下AT组的肝组织肿胀明显,而HP组无明显改变.透射电镜下AT组肝细胞出现典型的凋亡征象,而HP组肝细胞无明显凋亡形态;定量分析线粒体的面积、周长及直径均与AT组差异有统计学意义(P＜0.05),HP组线粒体损伤程度较AT组为轻.结论 HP对移植所导致的肝细胞凋亡有明显的抑制作用;它可能系通过上调Bcl-2蛋白的表达、减轻线粒体损伤途径而发挥其保护作用.     

N-乙酰半胱氨酸预处理对肝脏缺血再灌注损伤的保护作用

目的 探讨N-乙酰半胱氨酸(N-acetylcysteine,NAC)预处理对肝脏缺血再灌注损伤的保护作用.方法 18 例肝血管瘤手术患者随机分为3 组(n=6):对照组(C0)、NAC 预处理组(PR)和NAC 后处理组(PO),其中PR和PO 组分别于肝门阻断前和开放后给予NAC 120 mg/kg.三组患者均于切皮前(T0)、肝门阻断开放后1 h(T1)及6 h(T2)抽血检测谷丙转氨酶活性(ALT)和谷草转氨酶(AST)水平,并于肝门阻断开放后1 h(T1)取肝组织测定丙二醛(MDA)含量及核因子κB(NF-κB)活性.结果 肝门阻断开放后1 h(T1)和6 h(T6),各组的ALT 和AST 水平均明显高于肝门阻断前(T0)水平(P<0.01);与对照组相比,PR 和PO 组的MDA 含量在肝门阻断开放后1 h(T1)明显减少(P<0.01),但NF-κB 的活性仅在PR 组显示有明显的降低(P<0.01).结论 NAC 预处理可有效防治肝脏缺血再灌注损伤,其机制与抑制再灌注后NF-κB 的启动激活有关.     

低氧预适应对自体原位肝移植大鼠p-JNK蛋白表达的影响

目的：探讨低氧预适应（HP）对大鼠自体肝移植后p-JNK蛋白表达的影响及其保护作用。方法：采用改良的自体肝移植模型,将SD大鼠随机分为正常对照（NC）组、自体肝移植（AT）组和＋AT（HP）组,每组32只。HP组术前用8%氮氧混合气体处理90min。分别于术后1、2、12和24h处死各组大鼠,每个时相点处死8只,取肝脏标本,抽血检测肝功能,免疫组织化学方法测定大鼠肝组织磷酸化的JNK（p-JNK）蛋白、细胞色素C的表达,并在透射电子显微镜下观察各组肝细胞形态学改变。结果：HP组血清ALT和AST水平显著低于AT组,差异有统计学意义（P〈0.05）;HP组各时相点的肝功能明显优于AT组;HP组各时相点p-JNK蛋白、细胞色素C的表达明显低于AT组,差异有统计学意义（P〈0.05）。透射电子显微镜下AT组肝细胞出现典型的凋亡征象,而HP组肝细胞无明显凋亡形态。结论：HP对移植所导致的肝细胞凋亡有明显的抑制作用;其机制可能是通过抑制JNK信号通路的持续激活,下调p-JNK蛋白的表达、抑制线粒体细胞色素C的释放而发挥其保护作用。     

潘生丁对实验大鼠肝缺血再灌注损伤的影响

目的探讨潘生丁对肝缺血再灌注损伤的保护作用。方法建立大鼠局部肝脏缺血再灌注模型。将24只健康雄性Wistar大鼠随机分为假手术组、缺血再灌注组、潘生丁预处理组,观察各组血浆肝酶及透明质酸(HA)水平变化和肝组织中丙二醛(MDA)、内皮素(ET-1)含量,并行肝组织病理形态学检查。结果与缺血肝组织相比,潘生丁预处理组肝酶的漏出、血浆HA水平及肝组织中MDA、ET-1的含量明显降低(P<0.01)。肝组织病理学损伤亦明显减轻。结论潘生丁预处理可明显改善肝微循环,减轻肝缺血再灌注损伤。药物预处理可为临床提供一种安全有效的预处理方法。     

Genistein预处理对大鼠肝缺血再灌注损伤的保护作用

目的：探讨genistein预处理对肝脏缺血再灌注损伤的保护作用及其机制。
方法：54只SD大鼠完全随机分成3组。A组为I/R组:70%热缺血60 min及再灌注;B组为I/R
 genistein（GST）预处理组;C组为假手术组。于成模后2,6,12 h取大鼠血清和肝组织,检测血清AST,ALT浓度和肝组织MDA浓度,免疫组化法检测肝组织casepase-3蛋白的表达,光镜下观察肝脏组织病理变化。
结果：与I/R组相比,genistein预处理血清中AST及ALT浓度明显降低,肝组织病理损伤明显减轻。肝组织casepase-3蛋白表达及MDA浓度显著降低（均P＜0.05）。
结论：genistein预处理对大鼠肝脏热缺血再灌注具有保护作用,其机制与清除氧自由基,抑制细胞凋亡有关。     

安氟醚和异氟醚对肝脏缺血/再灌注损害的影响

目的 研究安氟醚和异氟醚预处理对肝脏缺血／再灌注损害的影响。方法 １８只家兔随机分３组,对照组阻断肝劝脉和门静脉血流形成肝缺血４５ｍｉｎ,开放再灌注１２０ｍｉｎ;安氟醚和异氟醚预处理组,于缺血再灌注前分别吸入１．６８％安氟醚和１．１５％异氟醚２０ｍｉｎ和药物清除１０ｍｉｎ。结果 缺血前,三组血清谷丙转氨酶（ＡＬＴ）,谷草转氨酶（ＡＳＴ）、乳酸脱氢酶（ＬＤＨ）、碱性磷酸酶（ＡＬＰ）和谷氨酰酶（ＧＣＴ     

缺血预处理对肝脏缺血再灌注损伤保护作用的实验研究

在大鼠肝脏缺血再灌注模型基础上建立稳定的缺血预处理模型,观察其保护作用,结果发现预处理与缺血再灌注组相比比清肝酶（ＡＬＴ,ＡＳＴ）,透明质酸水平及肝组织局部ＭＤＡ含量均低,而组织ＡＴＰ含量及能荷值均高,形态学损伤改变轻;同时发现预处理组织腺苷含量明显高于缺血再灌注组。实验表明缺血预处理能有效减轻肝脏的缺血再灌注损伤,腺苷分子参与了这一保护机制。