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落新妇苷通过促进IL-10表达保护小鼠缺血再灌注损伤肝脏
慕宁1,王海梁1,傅宏1,林峰1,施晓敏1,邹绍武1,王全兴2,傅志仁1*,傅志仁
0
(1.第二军医大学长征医院全军器官移植研究所,上海 200003*2.第二军医大学基础部免疫学教研室,免疫学研究所,上海 200433;第二军医大学附属长征医院)
摘要:
目的:观察落新妇苷对肝脏热缺血再灌注损伤的保护作用,对其机制进行初步探讨。方法:C57BL/6小鼠随机分为4组:假手术组(Sham)、模型组(I/R)、落新妇苷小剂量(10 mg/kg)干预组和落新妇苷大剂量(40 mg/kg)干预组。缺血前24 h和1 h干预组小鼠腹腔注射分别给予10或40 mg/kg的落新妇苷,建立肝左、中叶70%部分肝缺血再灌注模型,模型组和假手术组给予同样体积的生理盐水。小鼠肝脏左叶缺血90 min、再灌注6 h后各实验组采集血液和肝脏组织样本。检测血清中丙氨酸转氨酶(ALT)的活性作为肝功能损伤的指标,同时用ELISA检测肝组织中MPO含量。观察肝脏组织病理学改变。Western印迹检测肝组织中IL-10蛋白含量,半定量RT-PCR检测IL-10 mRNA表达情况。结果:落新妇苷干预能有效降低部分肝脏热缺血再灌注小鼠血清ALT水平,能有效降低缺血肝脏中MPO含量,改善肝组织病理表现。干预组肝组织中IL-10蛋白表达与I/R模型对照组比较均渐次升高,与半定量RT-PCR结果相符(小剂量干预组P<0.05;大剂量干预组P<0.01)。结论:落新妇苷干预能减轻小鼠肝脏热缺血再灌注损伤后的炎症反应,有效改善肝功能和肝脏病理损害;机制可能在于其能促进缺血再灌注损伤肝组织中IL-10的高表达。
关键词:    再灌注损伤  落新妇苷  丙氨酸转氨酶  髓过氧化物酶  白细胞介素-10
DOI:10.3724/SP.J.1008.2008.01429
投稿时间:2008-06-06修订日期:2008-10-10
基金项目:
Protective effect of astilbin on ischemia/reperfusion-induced liver injury through increasing IL-10 expression
MU Ning1,WANG Hai-liang1,FU Hong1,LIN Feng1,SHI Xiao-min1,ZOU Shao-wu1,WANG Quan-xing2,FU Zhi-ren1*,Fu Zhi-ren
(1.PLA Institute of Organ Transplantation,Chengzheng Hospital,Second Military Medical University,Shanghai 200003,China*2.Department Immunology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433)
Abstract:
Objective:To investigate the protective effect of astilbin on warm ischemia/reperfusion-induced liver injury and to study the related mechanism.Methods:C57BL/6 mice were randomly divided into four groups (n=8): sham-operated group (Sham),model control group (I/R),low dose astilbin treatment group (10 mg/kg) and high dose astilbin (40 mg/kg) treatment group. Mice in the two treatment groups were intraperitoneally injected with astilbin 24 hours and one hour before ischemia. Then 70% hepatic ischemia/reperfusion model (the left and middle hepatic lobe) was established. Mice in the I/R model control group and the sham operation group were administered with the same volume of normal saline. The blood sample and liver tissue samples were collected 90 min after ischemia and 6 h after reperfusion. Serum ALT activity was detected as an indicator of liver function damage and the content of MPO in liver tissues were detected by ELISA. The pathological changes of the liver were observed. The expression of IL-10 in liver tissues was detected by Western blotting and the expression of IL-10 mRNA was detected by semi-quantitative RT-PCR.Results:Astilbin treatment can effectively lower the serum ALT level and MPO level in the liver tissues in some I/R mice. It could also improve the pathological manifestations of the liver. Compared with the I/R model control group,IL-10 protein levels gradually increased in the two treatment groups,which was consistent with the result of RT-PCR (low dose group P<0.05; high dose group,P<0.01).Conclusion:Astilbin can effectively reduce the inflammatory response after liver warm ischemia-reperfusion induced injury,effectively improve the mouse liver function and pathological damage,which might be related to the upregulation of IL-10 expression in the liver tissues.
Key words:  liver  reperfusion injuries  astilbin  alanine transaminase  myeloperoxidase  iuterleukin-10