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目的 通过研究急性缺血缺氧及再灌注时胎鼠肾脏一氧化氮 (NO)的变化规律及与一氧化氮合酶 (NOS)的关系 ,初步探讨急性缺血缺氧及再灌注时肾损伤的发病机制。 方法 胎龄 2 1日胎鼠为研究对象 ,分为缺血缺氧 10min、3 0min及缺血缺氧 3 0min再灌注 3 0min、2h、6h和 2 4h组 ,应用硝酸还原酶法、免疫组化和RT PCR技术检测不同时间NO含量改变和NOS(内皮型eNOS和诱导型iNOS)活性及基因表达。 结果 随缺血缺氧时间延长NO水平明显降低 ,缺血 10min (2 2 .65± 4.0 1) ,缺血 3 0min(17.0 4± 2 .99)与假手术组 (2 5 .2 6± 3 .0 5 )相比 ,差异有统计学意义 (P <0 .0 1)。随再灌注时间延长NO水平呈双向改变 ,于再灌注 6h升高达高峰 (2 9.78± 2 .18) ,再灌注 2 4h仍维持较高水平 (2 9.45± 2 .78,P <0 .0 5 )。正常时eNOS和iNOS即位于近曲小管。随缺血缺氧及再灌注时间的延长 ,eNOSmRNA含量逐渐降低 ,eNOS光强度逐渐升高 ,活性逐渐减弱 ,与假手术组相比 ,差异有统计学意义 (P <0 .0 1) ;iNOSmRNA含量及光强度改变与之相反 ,于再灌注 6h后与假手术组差异有统计学意义 (P分别 <0 .0 5和 0 .0 1)。 结论 急性缺血缺氧及再灌注时胎鼠肾脏NO水平呈双相改变 ,参与肾损伤 ,其动态变化过程是NOS体系综合作用的结果
OBJECTIVE: To investigate the changes of nitric oxide (NO) and the relationship with nitric oxide synthase (NOS) in fetal rat kidney during acute ischemia, hypoxia and reperfusion, and to investigate the effects of acute ischemia, hypoxia and reperfusion on renal injury The pathogenesis. Methods Fetal gestation day 2 1 fetuses were divided into hypoxia and hypoxia 10 min, 30 min and reperfusion 30 min, 2 h, 6 h and 24 h after ischemia-reperfusion 30 min. Nitric acid reductase The changes of NO content and NOS (endothelial eNOS and inducible iNOS) activity and gene expression at different time were detected by RT-PCR and RT-PCR. Results The level of NO was significantly decreased with prolonged hypoxia / hypoxia time. There was no significant difference between the two groups (P <0.05) .0 5), the difference was statistically significant (P <0.01). The level of NO was changed bidirectionally with prolonged reperfusion time, reaching the peak at 6h after reperfusion (2 9.78 ± 2.18) and remained at a high level 24 h after reperfusion (2 9.45 ± 2.78, P <0.05 ). Normal eNOS and iNOS that is located in the proximal tubule. Compared with the sham-operated group, the eNOS mRNA level gradually decreased and the eNOS light intensity gradually increased, while the activity decreased gradually with the time of ischemia-hypoxia and reperfusion. The difference was statistically significant (P <0.01); iNOS mRNA On the contrary, the content and intensity of light changes were statistically different from those of sham operation group at 6h after reperfusion (P <0.05 and 0.01 respectively). Conclusions Acute ischemia, hypoxia and reperfusion have a biphasic change of NO level in fetal kidney, which is involved in renal injury. The dynamic change process is the result of the comprehensive effect of NOS system