GENE THERAPY OD ACUTE RENAL FAILURE BY CELL CYCLE-REGULATED GENES

通过细胞周期调控基因治疗急性肾衰竭

基本信息

批准号：
12671029
负责人：
TERADA Yoshio
金额：
$ 2.05万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671029/
关键词：
Wnt-4 acute renal failure cell cycle ischemia apoptosis proximal tubule proliferaion cyclin アデノウイルスサイクリン尿細管細胞 CGHP cGMP

项目摘要

We investigated the genes which play important roles in the regeneration of renal tubules during renal failure. Wnt-4 is known to be expressed in the mesonephric duct in embryonic development. It is tempting to speculate that the Wnt-4-β-catenin pathway contributes to the recovery from acute renal failure (ARF). In this study we used an in vivo model of ARF rats to clarify the significance of the Wnt-4-β-catenin pathway in ARF. ARF was induced by clamping the rat left renal artery for 1h. At 3, 6, 12, 24, 48, and 72 h after reperfusion, we extracted whole kidney homogenate and total RNA for examination by Western blot analysis and real-time RT-PCR. Wnt-4 mRNAand protein expression were strongly increased at 3-12 h and 6-24 h after ischemia, respectively. In immunohistological examination, Wnt-4 was expressed in the proximal tubules and co- expressed with aquaporin-1, GM130, and PCNA Cyclin Dl and cyclin A were expressed at 24-48 h after reperfusion. In addition, the overexpression of Wnt-4 and β- catenin promoted the cell cycle and increased the promoter activity and protein expression of cyclin Dl in LLC-PK1 cells. Taken together, these data suggest that the Wnt-4-β-catenin pathway plays a key role in the cell cycle progression of renal tubules in ARF. The Wnt-4-β-catenin pathway may regulate the transcription of cyclin Dl and control the regeneration of renal tubules in ARF.

我们研究了在肾衰竭期间肾脏块茎再生中起重要作用的基因。已知Wnt-4在胚胎发育中的嗅觉管道中表达。很容易推测WNT-4-β-catenin途径有助于从急性肾衰竭（ARF）中恢复。在这项研究中，我们使用了ARF大鼠体内模型来阐明ARF中Wnt-4-β-catenin途径的重要性。通过将大鼠左肾动脉夹住1H来诱导ARF。再灌注后3、6、12、24、48和72小时，我们提取了整个肾脏匀浆和总RNA，以通过蛋白质印迹分析和实时RT-PCR进行检查。缺血后3-12小时和6-24小时，Wnt-4 mRNAAND蛋白表达大大增加。在免疫组织学检查中，WNT-4在近端管中表达，并与Aquaporin-1，GM130和PCNA Cyclin dl和Cyclin A共表达，在再灌注后24-48小时表达。此外，Wnt-4和β-catenin的过表达促进了细胞周期，并增加了Cyclin DL在LLC-PK1细胞中的启动子活性和蛋白质表达。综上所述，这些数据表明WNT-4-β-catenin途径在ARF中肾管的细胞周期进程中起关键作用。 Wnt-4-β-catenin途径可以调节细胞周期蛋白DL的转录并控制ARF中肾小管的再生。