缺血后处理对大鼠肝脏一氧化氮合酶表达的影响及意义  

Effects of ischemic postconditioning on expression of liver nitric oxide synthase in rat and its significance

作　　者：曹能琦[1] 蒋维维[1] 刘养岁[1] 浦立勇[1] 宋德静[1] 孔连宝[1] 

机构地区：[1]南京医科大学第一附属医院肝脏移植中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第4期433-436,498,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生厅兴卫工程重点人才资助(RC2007058)

摘　　要：目的:探讨缺血后处理(ischemic postconditioning,IPO)对大鼠肝脏一氧化氮合酶(nitric oxide sythase,NOS)表达的影响及意义。方法:建立70%大鼠肝脏缺血再灌注模型,将32只雄性SD大鼠随机分为3组:假手术组(SO组,n=8)、缺血再灌注组(IR组,n=12)、缺血后处理组(IPO组,n=12),IPO组于再灌注恢复血流前,给予多次短暂复灌复停处理。再灌注3h后,比较各组血清谷丙转氨酶(ALT)、谷草转氨酶(AST)变化,硝酸还原酶法测定一氧化氮(NO)含量,逆转录-聚合酶链反应法(RT-PCR)检测肝脏组织内皮型NOS(eNOS)mRNA、诱导型NOS(iNOS)mRNA表达,蛋白免疫印迹法(Western blot)检测肝脏组织eNOS、iNOS蛋白表达,光镜观察组织病理学改变。结果:与SO组相比,其余两组血清ALT、AST、NO含量均明显升高(P<0.01),组织eNOS、iNOS表达均明显增强。与IR组相比,IPO组ALT、AST明显降低(P<0.01),NO明显升高(P<0.01),eNOS、iNOS表达增强。光镜下,IR组、IPO组呈现典型缺血再灌注损伤的病理学改变,但IPO组损伤较IR组轻。结论:缺血后处理能明显减轻大鼠肝脏缺血再灌注损伤,其机制可能与增强缺血再灌注肝脏组织中eNOS表达、提高血清NO含量有关。Objective：To investigate the effects of ischemic postconditioning on expression of liver nitric oxide synthase（NOS） in rat. Methods：The model of 70% hepatic ischemia reperfusion was constructed in SD male rat. Thirty two SD rats were randomly divided into three groups：sham group （SO group,8 rats）,ischemia reperfusion group （IR group,12 rats） and ischemic postconditioning group （IPO group,12 rats）. In group IPO,rats were treated as IR group,but at the onset of reperfusion,reflow was initiated with 10 s of full blood flow,followed by 10 s of re-occlusion,and repeated six times. After reperfusion for 3 hours,changes of ALT,AST and NO in different groups were compared. Reverse transcription-polymerase chain reaction and western blot were used to detct the expressions of NOS mRNA and protein respectively. Histopathological changes were observed by light microscope. Results：Compared with the SO group,ALT,AST and NO in serum were significantly higher （P 〈 0.01） than those in the other groups,and the mRNA and protein expressions of NOS were also obviously increased. Compared to the group IR,the levels of ALT,AST were significantly lower （P 〈 0.01）,while NO was significantly higher（P 〈 0.01） in the IPO group. The expressions of NOS mRNA and protein were increased in the IPO group. Under light microscope,the group IR and IPO both showed typical pathological changes of ischemic-reperfusion injury,but the injury was extenuated in the IPO group. Conclusion：Ischemic postconditioning can protect liver from ischemic-reperfusion injury. The mechanism might be related with the induction of eNOS in tissue and the increase of NO in serum.

关 键 词：缺血后处理 肝脏 缺血再灌注 诱导型一氧化氮合酶 内皮型一氧化氮合酶 

分 类 号：R657.3[医药卫生—外科学]