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目的:探讨胆道梗阻围手术期解除梗阻后肾脏水通道蛋白1的变化,以及应用小剂量多巴酚丁胺对围手术期肾水通道蛋白1表达的影响。方法:70只大鼠随机分为4组,分别为对照组10只、梗阻性黄疸组20只、多巴酚5μg组20只、多巴酚10μg组20只。实验组60只大鼠全身麻醉下建立梗阻性黄疸动物模型;对照组10只大鼠行假手术。7d后实验组再次全麻解除梗阻,而对照组再次行假手术。多巴酚丁胺组于胆道再通手术时于股静脉套管针穿刺泵入多巴酚丁胺。分为5μg/(kg.min)和10μg/(kg.min)两组;而假手术组及梗阻性黄疸组20只大鼠泵入0.9%生理盐水,2h后结束药物输注,并将各组动物随机均分为立即取材组和24h后取材组。将静脉血离心后将血清冻存检测肝功能、肾功能;部分肾脏标本冻存,RT-PCR法半定量检测水通道蛋白1aquaporin1mRNA相对表达;蛋白印迹Western blotting法半定量检测aquaporin1蛋白表达。结果:解除梗阻早期鼠血清胆红素水平下降,同时BUN及Cr未见明显改变。通过Western印迹对肾aqp1表达测定发现:多巴酚丁胺5μg组较10μg组aqp1表达更接近正常对照组,同时梗阻性黄疸组aqp1表达较对照组明显下降。而RT-PCR法检测aqp1mRNA结果与蛋白表达相反,且均存在明显差异。结论:应用小剂量多巴酚丁胺对于增加肾近曲小管上皮细胞aquaporin1表达具有积极作用,可能因而改善梗阻性黄疸围手术期肾功能。
OBJECTIVE: To investigate the changes of renal aquaporin 1 after perioperative obstruction of biliary obstruction and the effect of low-dose dobutamine on renal aquaporin-1 expression. Methods: Seventy rats were randomly divided into four groups: control group (n = 10), obstructive jaundice group (n = 20), pentobarbital 5μg group (n = 20) and doxate 10μg group (n = 20). 60 rats in the experimental group were established animal models of obstructive jaundice under general anesthesia; 10 rats in the control group underwent sham operation. The experimental group again after general anesthesia 7d lifting obstruction, while the control group re-sham surgery. Dobutamine group in the biliary recanalization surgery in the femoral vein puncture pump dobutamine. Divided into 5μg / (kg.min) and 10μg / (kg.min) two groups; sham operation group and obstructive jaundice group 20 rats were pumped 0.9% saline, 2h after the end of drug infusion, and each Groups of animals were randomly divided into immediate access group and 24 hours after the material group. The venous blood was centrifuged and the serum was frozen to detect liver function and renal function. Some kidney samples were frozen and the expression of aquaporin1 protein was detected by semi-quantitative RT-PCR and Western blotting respectively. Results: Serum bilirubin level was decreased in the early stage of obstruction, and no obvious changes were found in BUN and Cr. Western blot analysis of kidney aqp1 expression found: 5μg dobutamine group than 10μg aqp1 expression closer to the normal control group, and obstructive jaundice aqp1 expression was significantly lower than the control group. RT-PCR detected aqp1mRNA and protein expression contrary, and there are significant differences. Conclusion: The use of low-dose dobutamine has a positive effect on the aquaporin1 expression in renal proximal tubule epithelial cells, which may result in the improvement of perioperative renal function in patients with obstructive jaundice.