肾缺血-再灌注后大鼠肾组织一氧化氮的代谢研究

目的：探讨肾I-R后大鼠肾组织NO的代谢。方法：建立大鼠肾I-R模型，用硝酸还原酶法测定NO含量，SP试剂盒免疫组织化学法检测肾组织cNOS、iNOS蛋白定位表达，Western blot检测肾组织cNOS、iNOS蛋白半定量表达。结果：（1）肾I-R后大鼠肾脏NO代谢异常，入肾血、出肾血NO^2-／NO^3-含量增多；同时肾组织NO^2-／NO^3-含量和尿液NO^2-／NO^3-排出率也有增高（P＜0．05）；（2）肾I-R后大鼠肾组织cNOS表达增多。以I-R1h最明显；iNOS则在I-R5h表达增多；（3）I-R1h和I-R5h大鼠肾小球内cNOS阳性信号均明显增强；而肾小管内cNOS阳性信号I-R1h显著减弱；I-R1h可见髓袢升枝和降枝粗段的肾小管有微弱的iNOS表达。而I-R5h肾小管的iNOS阳性信号明显增强。结论：肾I-R导致了大鼠肾脏cNOS、iNOS蛋白表达改变，从而使NO代谢异常。

Study on Metabolism of Renal Nitric Oxide in Rat with Renal Ischemia- reperfusion

Objective: To investigate metabolism of renal nitric oxide in rat with renal ischemia-reperfusion.Methods: In the rat model of renal ischemia-reperfusion,NO~(2-)/ NO~(3-) was determined by nitrate reductase and renal cNOS protein was detected by ABC immunohistochemistry and Western blot.Results:(1) Metabolism of renal nitric oxide was Abnormal in rat with renal ischemia-reperfusion: the NO~(2-)/NO~(3-)content of IRB,ORB was remarkably increased(P<0.01),meanwhile the NO~(2-)/NO~(3-)content of kidney and urine also increased(P<0.05).(2) Expression of cNOS was strengthened especially in I-R1?h,but the iNOS signal was stronger in I-R5?h rats.(3) 140?kDa cNOS protein appeared in the kidney of sham.For IR1?h and I-R5?h rats,I-R1?h was the most distinctive,next was I-R5?h.There was no 135?kDa iNOS protein in sham,a little in I-R1?h,a great deal in I-R5?h.Conclusion: cNOS and iNOS proteins were changed in renal I-R rats,which results in the abnormal of NO metabolism.