小鼠肾脏缺血再灌注时肾组织角蛋白家族基因表达的变化  

Changes in expression of keratin genes in renal tissues during renal ischemia-reperfusion injury in mice

作　　者：刘宇琦[2] 严欢[1] 仓静[1] 薛张纲[1] 王浩[1] 

机构地区：[1]复旦大学附属中山医院麻醉科,上海市200032 [2]复旦大学附属妇产科医院麻醉科

出　　处：《中华麻醉学杂志》2017年第1期104-107,共4页Chinese Journal of Anesthesiology

摘　　要：目的 探讨小鼠肾脏缺血再灌注时肾组织角蛋白家族基因表达的变化.方法 野生型雄性C57/B6小鼠6只,50日龄,体重20～30 g,采用随机数字表法分为2组（n=3）：假手术组（Sham组）和缺血再灌注组（I/R组）.采用夹闭右侧肾动脉与肾静脉1h后再恢复灌注,并切除左侧肾脏的方法建立小鼠肾脏缺血再灌注模型.再灌注24 h时经左心室采血,比色法检测血清肌酐及尿素氮浓度;取右侧肾组织,HE染色法观察病理学结果;采用qRT-PCR检测肾组织Kim-1、Ngal的mR-NA表达水平;采用基因表达谱芯片分析技术检测肾组织角蛋白家族基因的表达水平,筛选出的差异基因进一步通过qRT-PCR验证.结果 与Sham组相比,I/R组血清肌酐和尿素氮浓度明升高,肾组织Kim-1、Ngal的mRNA表达上调（P＜0.05）,肾小管病理学损伤加重.基因表达谱芯片检测结果显示角蛋白家族基因中仅角蛋白20基因（表达上调）为差异表达基因（P＜0.05）,qRT-PCR验证检测结果与基因表达谱芯片检测结果一致.结论 小鼠肾脏缺血再灌注损伤时肾组织角蛋白20基因表达上调,该变化可能参与了肾脏缺血再灌注损伤时的内源性保护机制.Objective To investigate the changes in the expression of keratin genes in renal tissues during renal ischemia-reperfusion （I/R） injury in mice.Methods Six wild type male C57/B6 mice,aged 50 days,weighing 20-30 g,were divided into 2 groups （n=3 each） using a random number table：sham operation group （Sham group） and I/R group.Right renal arteries and veins were clamped for 1 h followed by reperfusion,and the left kidneys were removed to establish the model of renal I/R injury.At 24 h of reperfusion,blood samples were collected from the left ventricle for determination of serum creatinine and urea nitrogen concentrations by colorimetric method.The right kidney specimens were obtained for pathologic examination and for determination of the expression of kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin mRNA （by quantitative real-time polymerase chain reaction [qRT-PCR]） and keratin genes （by Affemetrixc DNA microarray）.The differentially expressed genes identified were further confirmed by qRT-PCR.Results Compared with Sham group,the serum creatinine and urea nitrogen concentrations were significantly increased,the expression of kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin mRNA was up-regulated （P〈0.05）,and the damage to the renal tubules was aggravated in I/R group.The results of microarray analysis showed that only keratin 20 gene （the expresion was up-regulated） was the differentially expressed gene （P〈0.05）,and the results measured by qRT-PCR were consistent with those measured by Affemetrixc DNA microarray.Conclusion Keratin 20 gene expression in renal tissues is up-regulated during renal I/R injury in mice,and the change may be involved in the endogenous protective mechanism during renal I/R injury.

关 键 词：角蛋白质类 再灌注损伤 肾 

分 类 号：R692[医药卫生—泌尿科学]