CAJ | 학술논문

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。
목적관찰결혈예괄응（IPC）대지체결혈재관주（LIR）후소장손상적보호효응，탐토일양화담（NO）재해과정중적작용궤제。방법웅성Wistar대서52지，수궤분위4조（n＝13）：가수술（Sham）조，지체결혈재관주（IR）조，결혈예괄응（IPC）조，Nω-초기-L-정안산갑지처리（L-NAME）조，측정각조대서혈장화소장조직이알양화매（DAO）、NO함량，혈장이류청산형광소표기적지다당（FITC-LPS）농도，소장조직병이철（MDA）、배린산조감（cGMP）적함량；채용면역조직화학법검측단백P-선택소（P-selection）화조망단백매（Caspase-3）적표체정황，이용원위탈양핵당핵감산말단전이매개도적결구말단표기법（TUNEL）검측각조소장조직세포적조망정황，사용자동도상분석계통대기결과진행정량분석。결과 IPC조여IR조비교，혈장DAO화FITC-LPS명현강저，NO승고（P＜0．01），소장조직NO、DAO화cGMP농도승고，MDA농도하강（P＜0．01），소장조직점막상피세포P-selection、Caspase-3적표체명현하강，AI강저（P＜0．01）；L-NAME조여 IPC조비교，혈장NO농도명현하강，DAO 화 FITC-LPS명현승고（P＜0．01），소장조직NO、DAO화cGMP농도명현하강，MDA농도승고（P＜0．01），P-selection、Caspase-3표체균명현상조，AI명현승고（P＜0．01）。결론 IPC가명현개선LIR후소장점막장통투성승고、조직지질과양화손상급장세포조망， NO삼여IPC적보호효응。
Objective 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 Nω-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 cGMP 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 cGM P 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 .