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缺血预处理对肝脏缺血再灌注损伤的保护机理研究

Mechanisms of preconditioning against ischemia reperfusion injury in rats liver

【作者】 王占明

【导师】 吴金生;

【作者基本信息】 第四军医大学 , 外科学(普外), 2001, 硕士

【摘要】 目的:研究缺血预处理对大鼠肝脏缺血再灌注损伤保护作用机制;绘制缺血再灌注后0—48h各损伤指标与抗损伤因素的变化曲线,并与缺血预处理后的指标变化进行对比;预处理的延迟保护效应与热休克蛋白的关系,预处理影响下热休克蛋白70mRNA表达水平的变化。方法:建立大鼠肝脏缺血再灌注模型并进行缺血预处理,实验分组如下①假手术组(Sham-operation, S组)、②缺血再灌注组(Ischemic Reperfusion, I/R组)、③缺血预处理组(IschemicPreconditioning, PC组)、④预处理加左旋硝基精氨酸组(Preconditioning+L-NNA, PC+L组),各组再灌注后检测血清丙氨酸转氨酶(ALT)及丙二醛(MDA)含量、热休克蛋白70(HSP70)免疫组化染色及mRNA检测,肝组织石蜡切片HE染色及电镜观察;结果:PC组ALT及MDA含量明显低于I/R组(p<0.01),肝组织损伤程度明显减轻;PC+L组在0—3h阶段ALT和MDA含量与I/R组无显著区别(p>0.05),而在6—48h阶段则显著低于I/R组(p<0.01)但仍高于PC组(p<0.05)。HSP70免疫组化染色:PC组、PC+L组3—48h染色阳性率逐渐增多并显著高于I/R组(p<0.01); 第四军医大学98年级硕士学位论文HSP70W表达逐渐增高,6一 12h 最高,其 PC组、PC+L组显著高于IR组一叩刀1人结论:缺血预处理对大鼠肝脏缺血再灌注损伤具有明显保护作用,0—3h为早期保护效应,可能为NO所触发或介导,6—48h为延迟保护效应(LateIschemic Preconditioning),HSP 70可能是导致延迟保护效应的重要因素。

【Abstract】 Objective: To study late preconditioning(PC) against ischemia/reperfusion(I/R) injury in rats liver. Methods 168 SD rats were divided into four groups: (1) S group: n=42,sham-operated control; (2)I/R group:n=42,Subjected to ischemic for 70-minute ischemia; (3) PC group: n=42,Two cycles of 5 min of ischemia followed by 5 min of reperfusion was performed before I/R;(4)PC+L group: n=42, L-NNA was administered before preconditioning then 70min ischemia. Followed by continuous reperfusion, serum ALT MDA NO NOS GSH GST and GSH-PX were examined in all groups during 0-48h; HSP 70 in the liver tissue were quantitatively analyzed and located by immunocytochemical technique. HSP 70 mRNAwere analyzed by PAGE Results Serum ALT and MDA in either I/R and PC+L group were markedly increased than in PC group (P<0.01) at any point after reperfusion.There is not significant difference between I/R group and P+L group during 0-3h(p>0.05) but during 6-48h(p<0.01). ALT and MDA were increased in PC+L vs PC group(p<0.05) and markedly decreased than I/R group during 6-48h(p<0.01). Expresstion of HSP 70 in PC and P+L group is gradually increased higher than in I/R group during 6-48h (P<0.01) ,so as HSP70 mRNA during l-12h.(p<0.01).Conc!usions: NO and HSP 70 might be participate in the protection against liver ischemia/reperfusion injury and NO might be protect liver in early preconditioning and HSP 70 might be associate with late preconditioning.

  • 【分类号】R657.3
  • 【下载频次】108
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