小鼠肝脏部分缺血再灌注损伤模型的建立
소서간장부분결혈재관주손상모형적건립
Establishment of mice model of partial hepatic ischemia reperfusion injury
  • 万方数据
    中华消化外科杂志 중화소화외과잡지
    CHINESE JOURNAL OF DIGESTIVE SURGERY
    2013年 9期 703-707 ,共5页

    麻勇%汪大伟%刘连新%王建奇%王继洲%潘华洋%潘尚哈%姜洪池

    마용%왕대위%류련신%왕건기%왕계주%반화양%반상합%강홍지

    肝脏%缺血再灌注损伤%小鼠%模型 肝髒%缺血再灌註損傷%小鼠%模型
    간장%결혈재관주손상%소서%모형
    Liver%Ischemia reperfusion injury%Mouse%Model
    目的 建立小鼠肝脏部分缺血再灌注损伤模型并分析损伤评估指标的变化趋势.方法 采用96只7～8周龄的纯系C57BL/6雄性小鼠作为研究对象,建立70％肝脏缺血再灌注损伤模型.按照缺血时间将小鼠分为假手术组和缺血30、60、90 min组,每组24只.各组小鼠分别于再灌注后6、12、24和48 h处死.通过检测血清ALT、AST、TNF-α、IL-6和巨噬细胞炎性蛋白-2(MIP-2)水平以及病理组织学评分、细胞凋亡指数等方法评估各组小鼠肝组织的损伤情况.两独立样本比较采用t检验.结果 术后88只小鼠存活,8只死亡,造模成功率为91.7％ (88/96).假手术组、缺血30、60、90 min组ALT水平分别为(35±24) U/L、(1703±442) U/L、(5133±681) U/L和(8233±808) U/L,缺血30、60、90 min组ALT水平显著高于假手术组(t=6.54,12.97,17.56,P＜0.05)；AST水平分别为(87±28) U/L、(2667 ±451) U/L、(6333±778)U/L和(9967±1168) U/L,缺血30、60、90 min组AST水平显著高于假手术组(t=9.89,13.89,14.65,P＜0.05)；TNF-α水平分别为(14 ±5) μg/L、(83±14) μg/L、(133±17) μg/L和(202±21) μg/L,缺血30、60、90 min组TNF-α水平显著高于假手术组(t=7.78,11.82,15.34,P＜0.05)；IL-6水平分别为(32 ±9) μg/L、(493±168) μg/L、(844±166) μg/L和(1345±198) μg/L,缺血30、60、90min组IL-6水平显著高于假手术组(=4.74,8.46,11.48,P＜0.05)；MIP-2水平分别为(37±11) μg/L、(102±35) μg/L、(177±32)μg/L和(279±50) μg/L,缺血30、60、90 min组MIP-2水平显著高于假手术组(t=3.05,7.28,8.19,P＜0.05)；细胞凋亡指数分别为1.7％±2.1％、22.7％±8.6％、54.3％±11.2％和76.3％±14.8％,缺血30、60、90 min组细胞凋亡指数显著高于假手术组(t=4.10,8.04,8.63,P＜0.05).在缺血时间相同的情况下,随着再灌注时间的延长,各监测指标呈“抛物线”样变化趋势.结论 小鼠肝脏部分缺血再灌注损伤模型能较好地反映小鼠肝组织的损伤情况.随着缺血时间的延长,小鼠肝脏的缺血再灌注损伤程度逐渐加重；随着再灌注时间的延长,ALT、AST、TNF-α、IL-6、MIP-2以及病理组织学评分和细胞凋亡指数均呈现“抛物线”样变化趋势.
    목적 건립소서간장부분결혈재관주손상모형병분석손상평고지표적변화추세.방법 채용96지7～8주령적순계C57BL/6웅성소서작위연구대상,건립70％간장결혈재관주손상모형.안조결혈시간장소서분위가수술조화결혈30、60、90 min조,매조24지.각조소서분별우재관주후6、12、24화48 h처사.통과검측혈청ALT、AST、TNF-α、IL-6화거서세포염성단백-2(MIP-2)수평이급병리조직학평분、세포조망지수등방법평고각조소서간조직적손상정황.량독립양본비교채용t검험.결과 술후88지소서존활,8지사망,조모성공솔위91.7％ (88/96).가수술조、결혈30、60、90 min조ALT수평분별위(35±24) U/L、(1703±442) U/L、(5133±681) U/L화(8233±808) U/L,결혈30、60、90 min조ALT수평현저고우가수술조(t=6.54,12.97,17.56,P＜0.05)；AST수평분별위(87±28) U/L、(2667 ±451) U/L、(6333±778)U/L화(9967±1168) U/L,결혈30、60、90 min조AST수평현저고우가수술조(t=9.89,13.89,14.65,P＜0.05)；TNF-α수평분별위(14 ±5) μg/L、(83±14) μg/L、(133±17) μg/L화(202±21) μg/L,결혈30、60、90 min조TNF-α수평현저고우가수술조(t=7.78,11.82,15.34,P＜0.05)；IL-6수평분별위(32 ±9) μg/L、(493±168) μg/L、(844±166) μg/L화(1345±198) μg/L,결혈30、60、90min조IL-6수평현저고우가수술조(=4.74,8.46,11.48,P＜0.05)；MIP-2수평분별위(37±11) μg/L、(102±35) μg/L、(177±32)μg/L화(279±50) μg/L,결혈30、60、90 min조MIP-2수평현저고우가수술조(t=3.05,7.28,8.19,P＜0.05)；세포조망지수분별위1.7％±2.1％、22.7％±8.6％、54.3％±11.2％화76.3％±14.8％,결혈30、60、90 min조세포조망지수현저고우가수술조(t=4.10,8.04,8.63,P＜0.05).재결혈시간상동적정황하,수착재관주시간적연장,각감측지표정“포물선”양변화추세.결론 소서간장부분결혈재관주손상모형능교호지반영소서간조직적손상정황.수착결혈시간적연장,소서간장적결혈재관주손상정도축점가중；수착재관주시간적연장,ALT、AST、TNF-α、IL-6、MIP-2이급병리조직학평분화세포조망지수균정현“포물선”양변화추세.
    Objective To establish mice model of partial hepatic ischemia reperfusion (IR) injury and investigate the variation trend of injury evaluation indexes.Methods Ninety-six male C57BL/6 mice (aged 7-8 weeks) were divided into 4 groups according to the ischemia time to construct hepatic IR model:sham operation group (24 mice),IR 30 minutes group (24 mice),IR 60 minutes group (24 mice) and IR 90 minutes group (24 mice).The mice in each group were sacrificed at hour 6,12,24,48 after IR.The injury of the hepatic tissues in the 4 groups was analyzed based on detecting the levels of alanine transaminase (ALT),aspartate transaminase (AST),tumor necrosis factor-α (TNF-ot),interleukin-6 (IL-6),macrophage inflammatory protein-2 (MIP-2),the histopathological grading and apoptosis index.All data were analyzed using the t test.Results Eight mice died,and the success rate of model construction was 91.7％ (88/96).The levels of ALT in the sham operation group,IR 30,60 and 90 minutes groups were (35 ±24)U/L,(1703 ±442)U/L,(5133 ±681)U/L and (8233 ±808) U/L,respectively.The levels of ALT in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =6.54,12.97,17.56,P ＜ 0.05).The levels of AST in the sham operation group,IR 30,60 and 90 minutes groups were (87 ± 28) U/L,(2667 ± 451) U/L,(6333 ± 778) U/L,(9967 ±1168) U/L,respectively.The levels of AST in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =9.89,13.89,14.65,P ＜ 0.05).The levels of TNF-α in the sham operation group,IR 30,60 and 90 minutes groups were (14 ± 5) μg/L,(83 ± 14) μg/L,(133 ± 17) μg/L,(202 ± 21) μg/L,respectively.The levels of TNF-α in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =7.78,11.82,15.34,P ＜ 0.05).The levels of IL-6 in the sham operation group,IR 30,60 and 90 minutes groups were (32 ± 9) μg/L,(493 ± 168) μg/L,(844 ± 166) μg/L,(1345 ± 198) μg/L,respectively.The levels of IL-6 in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =4.74,8.46,11.48,P ＜0.05).The levels of MIP-2 in the sham operation group,IR 30,60 and 90 minutes groups were (37 ± 11) μg/L,(102 ± 35) μg/L,(177 ± 32) μg/L,(279 ± 50) μg/L,respectively.The levels of MIP-2 in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =3.05,7.28,8.19,P ＜0.05).The apoptotic indexes in the sham operation group,IR 30,60 and 90 minutes groups were 1.7％ ± 2.1％,22.7％ ± 8.6％,54.3％ ± 11.2％ and 76.3％ ± 14.8％,respectively.The levels of apoptotic indexes in the IR 30,60 and 90 minutes groups were significantly higher than the sham operation group (t =4.10,8.04,8.63,P ＜ 0.05).Under the same ischemia time,the changes of ALT,AST,TNF-α,IL-6 and apoptosis index were presented as the parabolic style as the increase of the reperfusion time.Conclusions The mice model of hepatic IR injury could reflect the injured condition of the hepatic tissue of mice.Along with the time extension of ischemia,the severity of hepatic IR aggravated gradually.With the time extension of reperfusion,the levels of ALT,AST,TNF-α,IL-6,MIP-2,histopathological grading and apoptosis index change in a parabolic trend.
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