缺血预适应对大鼠肢体缺血再灌注后小肠损伤的保护效应及NO的作用  

Protective effect of ischemia preconditioning and the role of nitric oxide on the small intestines injury following hind limbs ischemia reperfusion in rat

作　　者：刘燕[1] 彭海兵[2] 张博[3] 周洪霞[1] 杨秀红[1] 张连元[1] 

机构地区：[1]河北联合大学基础医学院,河北唐山063300 [2]河北联合大学冀唐学院,河北唐山063300 [3]河北省沧州市中西医结合医院ICU,061001

出　　处：《重庆医学》2013年第21期2500-2502,2510,共4页Chongqing medicine

基　　金：河北省科技厅计划项目(12277793);唐山市科技局科研基金资助课题(12140209A-27)

摘　　要：目的观察缺血预适应(IPC)对肢体缺血再灌注(LIR)后小肠损伤的保护效应,探讨一氧化氮(NO)在该过程中的作用机制。方法雄性Wistar大鼠52只,随机分为4组(n=13):假手术(Sham)组,肢体缺血再灌注(IR)组,缺血预适应(IPC)组,Nω-硝基-L-精氨酸甲酯处理(L-NAME)组,测定各组大鼠血浆和小肠组织二胺氧化酶(DAO)、NO含量,血浆异硫氰酸荧光素标记的脂多糖(FITC-LPS)浓度,小肠组织丙二醛(MDA)、环磷酸鸟苷(cGMP)的含量;采用免疫组织化学法检测蛋白P-选择素(P-selection)和凋亡蛋白酶(Caspase-3)的表达情况,利用原位脱氧核糖核甘酸末端转移酶介导的缺口末端标记法(TUNEL)检测各组小肠组织细胞的凋亡情况,使用自动图像分析系统对其结果进行定量分析。结果 IPC组与IR组比较,血浆DAO和FITC-LPS明显降低,NO升高(P<0.01),小肠组织NO、DAO和cGMP浓度升高,MDA浓度下降(P<0.01),小肠组织黏膜上皮细胞P-selection、Caspase-3的表达明显下降,AI降低(P<0.01);L-NAME组与IPC组比较,血浆NO浓度明显下降,DAO和FITC-LPS明显升高(P<0.01),小肠组织NO、DAO和cGMP浓度明显下降,MDA浓度升高(P<0.01),P-selection、Caspase-3表达均明显上调,AI明显升高(P<0.01)。结论 IPC可明显改善LIR后小肠黏膜肠通透性升高、组织脂质过氧化损伤及肠细胞凋亡,NO参与IPC的保护效应。Ojective To observe the protection effect of ischemic preconditioning to small intestines injury after limbs ischemia reperfusion,and investigate the role of nitrogen monoxidum in the process. Methods 52 male Wistar rats were randomly divided in- to Sham group,limbs ischemia-reperfusion （IR） group, IPC ＋ IR group and Nco-nitro-L-arginine methylester （L-NAME） group. The value of diamine oxidase （DAO） and nitrogen monoxidum （NO） in plasma and gut tissue,the content of fluorescein isothiocya- nate lipopolysaccharide （FITC-LPS） in plasma, and the value of cyclic guanosine monophosphate （cGMP） and malondialehyde （MDA） in gut tissue were measured, respectively. The expression of P-selection and Caspase-3 were measured by immunohisto- chemistry. The apoptosis of intestinal cells was detected by terminal deoxynucleotidyl transferase--mediated dUTP nick end labe- ling （TUNEL） and the results were analysed quantitatively by automatic image analytical system. Results Compared with the LIR group,the values of DAO and FITC-LPS in plasma decreased obviously in the IPC group, and the level of NO in plasma increased obviously （P〈0. 01）. The values of DAO,NO and eGMP in gut tissue increased obviously,the level of MDA and the expression of Caspase-3 and the apoptosis index （AI） were significantly decreased in IPC group （P〈0.01）. Compared with the IPC group, the value of DAO and FITC-LPS were higher and the level of NO was lower in plasma of the L-NAME group （P〈0.01）. The values of DAO, NO and cGMP in gut tissue obviously decreased, the level of MDA significantly increased, and the expression of Caspase-3 and the AI significantly upregulated in L-NAME group than those in IPC group （P〈0.01）. Conclusion IPC can improve the small intestine injury and the apoptosis following hind limbs ischemia reperfusion which may correlated with the increase in the level of NO.

关 键 词：再灌注损伤 一氧化氮 细胞凋亡 缺血预适应 环磷酸鸟苷 

分 类 号：R965[医药卫生—药理学]