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内皮素(ET)是一种主要由内皮细胞分泌的肽类物质,具有强效的血管收缩作用,其中ET-1的缩血管作用最强。近年发现ET对缺血性急性肾衰竭的发生、发展起重要作用。本文研究肾缺血再灌注损伤对肾组织ET-1和ET受体基因mRNA转录调节的影响。材料和方法健康成年SD大鼠36只,苯巴比妥钠40rag/kg腹腔麻醉。腹部正中切口,游离左侧肾蒂,用无创动脉夹夹闭左肾动脉40分钟,然后再灌注0、2、6、24小时及2、7天,右肾作为对照(每一时间点n=6只)。异硫氰酸胍一步法提取肾组织总RNA,以1μg RNA为模板,使用ET-1、ET受体A和B(ET_A、ET_B)特异性引物,并以β-actin基因作为内参标准,半定
Endothelin (ET), a peptide secreted mainly by endothelial cells, has potent vasoconstriction, of which ET-1 has the strongest vasoconstrictor effect. In recent years, ET has been found to play an important role in the occurrence and development of ischemic acute renal failure. This study was designed to investigate the effects of renal ischemia-reperfusion injury on the transcriptional regulation of ET-1 and ET receptor mRNA in renal tissues. MATERIALS AND METHODS Thirty - six healthy adult Sprague - Dawley rats were anesthetized with phenobarbital at a dose of 40rag / kg. Abdominal midline incision, free left renal pedicle, with noninvasive artery clip occlusion of the left renal artery 40 minutes, and then reperfusion 0,2,6,24 hours and 2,7 days, the right kidney as a control (n = 6). One-step extraction of guanidine isothiocyanate was used to extract total RNA from renal tissues. The specific primers of ET-1, ET receptors A and B (ET_A, ET_B) were used with 1 μg of RNA as template. The β-actin gene was used as internal standard set