| 摘要: |
| 目的:探讨短时间多次缺血预处理(ischemic preconditioning,IP)对大鼠移植肝脏缺血再灌注损伤(ischemia-reperfusion injury,IRI)的保护作用。方法:采用SD大鼠原位肝移植动物模型,供肝冷保存120 min,无肝期14 min。54只SD大鼠随机分成5组：对照组(A组,n=6),不做肝脏移植手术；肝移植组(B组,n=12),供体大鼠肝脏4℃乳酸林格液保存2 h后,采用双袖套法行原位肝移植；缺血预处理肝移植组(C1、C2、C3组,每组12只)：将供肝进行缺血预处理后切除供体大鼠肝脏,4℃乳酸林格液保存2 h,行原位肝移植,根据阻断第一肝门5 min,开放再灌注5 min为1个循环,处理1～3个循环分别命名为C1、C2和C3组。术后检测各组血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)活性,每分钟胆汁量,免疫组化检测肝细胞Bcl-2表达,TUNEL法检测细胞凋亡,电镜观察细胞超微结构的改变。结果:术后B组血清ALT、AST、LDH活性明显高于A组,每分钟胆汁量明显低于A组(P<0.01)；C组ALT、AST、LDH活性虽明显高于A组(P<0.01),但比B组明显降低(P<0.05);随着IP次数增加,C1、C2、C3组ALT、AST、LDH活性递减,而每分钟胆汁量逐渐增加。细胞形态学检查发现,与B组比较,C组供肝组织细胞结构改变较小,Bcl-2表达增加,凋亡指数降低(P<0.01或P<0.05)。结论:IP对大鼠供肝缺血再灌注损伤具有保护作用,短时间多次IP对肝脏的保护作用更强。 |
| 关键词: 再灌注损伤 缺血预处理 肝移植 |
| DOI：10.3724/SP.J.1008.2008.01438 |
| 投稿时间：2007-07-10 |
| 基金项目:镇江市科技计划资助(SH2005037). |
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| Repeated ischemic preconditioning in a short time protects rat liver graft from ischemic reperfusion injury |
| ,PANG Li-qun1,FANG Ping-cheng2,LIU Da-biao 2 ,ZHENG Jian-ming3* |
| (1.Department of General Surgery,Affiliated Huaian Hospital of Xuzhou Medical College,Huaian 223002,China* 2.Department of General Surgery,Fourth Hospital of Jiangsu University,Zhenjiang 212000*3.Department of Pathology,Changhai Hospital,Second Military Medical University,Shanghai 200433) |
| Abstract: |
| Objective:To investigate the protective effect of repeated ischemic preconditioning (IP) in a short period on rat liver graft from ischemic reperfusion injury (IRI).Methods: Male Sprague-Dawley rats were used as donors and recipients for orthotopic liver transplantation.The period of cold preservation and anhepatic phase were 120 min and 14 min,respectively.Fifty-four rats were randomly divided into 5 groups.Control group (group A,n=6) received no operation of liver transplantation; liver transplantation group (group B,n=12) received orthotopic liver transplantation by the cuff technique with donor livers perserved for 2 h in 4℃ Ringer’s lactate solution before transplantation; and ischemic precondition group included three subgroups:group C1,C2,and C3(n=12).For group C,the grafts were harvested and then perserved for 2 h in 4℃ Ringer’s lactate solution after ischemic preconditioning,and then used for orthotopic liver transplantation.An IP cycle included 5 min interruption of the portal veins and hepatic arteries of liver,and reflow for 5 min.The C1,C2,and C3 animals were subjected to one,two and three circles,respectively.The activities of AST,ALT and LDH in the sera were examined and the capacity of bile per min was examined 6 hours after operation.The expression of Bcl-2 protein in hepatic tissue was determined with immunohistochemistry.Moreover,the hepatocellular apoptosis were detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and the changes of liver ultrastructure were observed under electron microscope.Results: The activities of ALT,AST,and LDH in group B were significantly higher and the capacity of bile per min was significantly lower than those in group A(P<0.01).The activities of ALT,AST,and LDH in group C were significantly higher than those of group A (P<0.01),but were significantly lower than those of group B (P<0.05),and decreased with the increase of IP cycle.Cell morphology examination showed that the changes of graft structure was slighter in group C than in group B,and the expression of Bcl-2 was increased and the apoptotic index was lowered (P<0.01 or P<0.05).Conclusion: Ischemic precondition can relieve IR-induced injury of the donor liver.The protective effect is more potent with repeated IP cycles in a short time. |
| Key words: reperfusion injury ischemic preconditioning liver transplantation |
