目的探讨阻断大鼠肝固有动脉建立肝缺血。再灌注损伤(IRI)模型的最佳灌注时间。方法sD大鼠48只,平均体重(200±25)g,按随机数字表法随机分为IR10、1、3、6、12、24、48h组和假手术(SO)组,每组各6只。IRI各组选择性阻断肝固有动脉1h后血流恢复,分别在血流恢复0、1、3、6、12、24、48h留取血和肝组织标本。SO组仅分离肝固有动脉,暴露第一肝门1h后取相应标本。观察各组大鼠血清ALT、AST水平,肝组织病理学改变,肝组织DNA片段化率,肝组织Ki.67表达水平。多组计量资料的比较采用单因素方差分析和LSD-t检验。结果IR10、1、3、6、12、24、48h组和SO组大鼠血清ALT水平分别为(53±25)、(85±20)、(96±L18)、(411±96)、(87±19)、(81±15)、(46±6)、(60±14)U/L。大鼠血清AST水平相应为(238±63)、(364±111)、(375±68)、(1291±511)、(800±87)、(854±218)、(484±219)、(248±94)U/L。IRI6h组血清ALT、AST水平最高(F=36.015,18.241;P〈0.05)。IRI6h组肝组织损伤程度最重。大鼠肝组织DNA片段化率相应为(7.5±1.5场、(9.2±2.2扬、(9.3±2.3)%、(12.6±2.4)%、(6.3±1.0)%、(5.4±0.9)%、(4.5±0.8)%、(4.5±1.1)%,6h组肝组织DNA片段化率最高(F=15.992,P〈0.05)。大鼠肝组织Ki.67表达相应为(3.5±1.4)、(5.6±1.8)、(8.7±2.3)、(13.7±2.4)、(15.2±1.2)、(20.5±2.2)、(31.8±2.5)、(2.4±L1.2)个/高倍视野,IRI48h组肝组织Ki-67表达水平最高(F=261.707,P〈0.05)。结论阻断大鼠肝固有动脉可以成功建立肝IRI模型,IRI最佳灌注时间为6h。
Objective To investigate the optimum reperfusion duration of liver ischemia- reperfusion injury (IRI) model by occluding the proper hepatic artery in rats. Methods Fourty-eight Sprague Dawley (SD) rats with mean weight of (200+25) g were randomly assigned to 8 groups by random number table method: IRI- 0, 1, 3, 6, 12, 24, 48 h and sham operation (SO) group with 6 rats in each group. The proper hepatic arteries in rats of IRI groups were selectively occluded for 1 h and then blood flow recovered. Samples of blood and liver tissues were collected at the time points of 0, 1, 3, 6, 12, 24, 48 h of reperfusion. In SO group, samples were collected after the proper hepatic artery was isolated and the first portal was exposed for 1 h. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), pathological changes, DNA fragmentation rates, and levels of Ki-67 expression of liver tissues were observed in each group. Measurement data of multiple groups were compared using one-way analysis of variance and LSD-t test. Results The levels of serum ALT in IRI- 0, 1, 3, 6, 12, 24, 48 h and SO group were (53±25), (85±20), (96±18), (411±96), (87±19), (81±15), (46±6), (60±14) U/L respectively. The levels of serum AST were (238±63), (364±111), (375±68), (1 291±511), (800~87), (854±218), (484±219), (248±94) U/L accordingly. The levels of serum ALT, AST in IRI 6 h group were the highest (F=36.015, 18.241; P〈0.05). The damage of liver tissues in IRI 6 h group was the most serious. The DNA fragmentation rates of liver tissues were (7.5±1.5)%, (9.2±2.2)%, (9.3±2.3)%, (12.6±2.4)%, (6.3±1.0)%, (5.4±0.9)%, (4.5±0.8)%, (4.5±1.1)% accordingly, which was the highest in IRI 6 h group (F=15.992, P〈0.05). The levels of Ki-67 expression of liver tissues were (3.5±1.4), (5.6±1.8), (8.7±2.3), (13.7±2.4), (15.2±1.2), (20.5±2.2), (31.8+