机构地区:[1]哈尔滨医科大学附属第一医院肝脏外科、省部共建教育部肝脾外科重点实验室,150001
出 处:《中华消化外科杂志》2013年第9期703-707,共5页Chinese Journal of Digestive Surgery
基 金:国家自然科学基金(81100305、81270527);中国博士后科学基金(2012M510990、2012M520769);黑龙江省博士后科学基金(LBH-Z11066);哈尔滨医科大学附属第一医院科研基金(2011BS001)
摘 要: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、90min组,每组24只。各组小鼠分别于再灌注后6、12、24和48h处死。通过检测血清ALT、AST、TNF一&、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、90min组A叫水平分别为(35±24)U/L、(1703±442)u/L、(5133±681)U/L和(8233±808)u/L,缺血30、60、90min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、90min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、90min组TNF-α水平显著高于假手术组(f=7.78,11.82,15.34,P〈0.05);IL-6水平分别为(32±9)μg/L、(493±168)陆∥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(t=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、90min组MIP-2水平显著高于假手术组(f=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、90min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-a (TNF-ct) , interleukin-a( IL-6), macrophage inflammatory protein-2 ( MIP-2 ) , the histopathologieal 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 thanthe sham operalhm 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/I,, (2667 ±451 )U/I,, (6333 ---778) U/L, (9967 ± 1168 ) U/I+, respe('tively. The levels of AST in tilt" 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-a in the sham operation group, 30, 60 and 90 nlinutes groups were ( 14 ± 5 ) ug/L, (83 ± 14) izg/L, ( 133 ± 17 ) txg/L, ( 202 ± 21 ) prespeetiwdy. The levels of TNV-ct 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
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