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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.

我们研究了在肾功能衰竭过程中对肾小管再生起重要作用的基因。已知WNT-4在胚胎发育过程中表达于中肾管。人们很容易推测WNT-4-β-连环蛋白途径有助于急性肾功能衰竭的恢复。在本研究中，我们使用了急性肾衰大鼠体内模型来阐明WNT-4-β-连环蛋白通路在急性肾衰中的意义。夹闭大鼠左肾动脉1h，建立ARF模型。分别于再灌注后3、6、12、24、48、72h提取全肾组织匀浆和总RNA，进行Western印迹分析和实时定量RT-PCR检测。WNT-4mRNA和蛋白的表达分别在缺血后3~12h和6~24 h显著增加。免疫组织化学显示，WNT-4表达于近端肾小管，并与水通道蛋白-1、GM130、增殖细胞核抗原、细胞周期蛋白、细胞周期蛋白A在再灌注后24~48h表达。此外，WNT-4和β-catenin的过表达促进了细胞周期，并增加了细胞周期蛋白Dl的启动子活性和蛋白表达。综上所述，这些数据提示WNT-4-β-连环蛋白通路在急性肾衰肾小管细胞周期进程中起关键作用。WNT-4-β-连环蛋白途径可能参与调控细胞周期蛋白Dl的转录，调控肾小管再生。