study on the changes in peritoneal ultrafiltration in peritoneal dialysis in relation to the regulation of the gene expression of peritoneal aquaporins

腹膜透析中腹膜超滤变化与腹膜水通道蛋白基因表达调控的研究

基本信息

批准号：
10671011
负责人：
HASEGAWA Hajime
金额：
$ 1.54万
依托单位：
Jikei University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

Recently, we have found that gene expression of peritoneal aquaporins (AQP1 and 4) was enhanced transitorily at the initial phase of continuous peritoneal dialysis (PD) in rats. To study the involvement of AQP in the ulltrafiltration loss occurred in long term CAPD patients, we designed long term experimental PD for 2 months in rats. Small catheter was settled in the abdominal cavity for exchanging dialysate and sampling.. Physiological saline (for control) or glucose-containing human-use dialysate(1.35% or 4.0%) were administered and exchanged twice a day. Daily monitored effluent volume (ultrafiltration volume) was increased in the initial phase and subsequently fixed, as observed in the previous experiment. Histological changes in the peritoneum such as mesothelium detachment or submesothelial fibrosis were not obvious. The effluent volume was gradually decreased after 5 weeks although we failed to obtain the statistical significance. Gene expression of peritoneal AQP1 and 4 after 8 … More weeks dialysis was inclined to be decreased comparing to that before dialysis (statistically insignificant) studied by RT-PCR combined with ELISA analysis. In situ hybridization study revealed that the signal accumulation in the capillary endothelium was less obvious after 8 weeks dialysis,but it was not significant. To confirm the involvement of AQP in the ultrafiltration failure in patients with long-term CAPD therapy, further study must be required. Next, to study the involvement of mechanical stretch in the induction of AQP gene expression, we designed in vivo and in vitro experiments. For in vivo experiment, small catheter with balloon was settled into the rats abdominal cavity to expand the peritoneum without dialysis. For in vitro experiment, mesothelial cells obtained by mechanical scratch of the peritoneum were cultured on the rubber plate, for subsequent application to 30% statistic stretch or 60 Hz cycled stretch. Peritoneum expansion in vivo generated transient enhancement of AQP gene expression at the day 3, which was similar pattern to that in experimental dialysis. However, there was no significant difference in in vitro experiement, suggesting that indirect effect of peritoneal expansion might be involved in the transient enhancement of AQP gene expression. Clinically, unexpected recovery from ultrafiltration failure in CAPD patients is sometimes experienced after few weeks discontinuation of PD. The peritoneal expansion induced AQP gene expression may be involved in the phenomenone. Less

最近，我们发现，在大鼠连续腹膜透析（PD）的初始阶段，腹膜水通道蛋白（AQP1和4）的基因表达得到了增强。为了研究长期CAPD患者的AQP参与无限液损失，我们在大鼠中设计了2个月的长期实验性PD。小导管定居在腹腔中以交换透析液和采样。生理盐水（用于对照）或含葡萄糖的人使用透析液（1.35％或4.0％）每天进行两次交换。如先前的实验中所观察到的那样，每日监测的有效体积（超滤体积）在初始阶段增加，随后固定。腹膜（例如中层脱离或服用纤维化）等腹膜的组织学变化并不明显。尽管我们未能获得统计显着性，但在5周后逐渐提高了有效体积。腹膜AQP1和4之后的腹膜AQP1的基因表达……与RT-PCR一起研究的透析（统计上无关）与ELISA分析相比，与透析（统计上无关紧要）相比，透析量较高。原位杂交研究表明，毛细血管内皮中的信号积累在透析8周后不太明显，但这并不重要。为了确认AQP参与长期CAPD治疗患者的超滤失败，必须进一步研究。接下来，为了研究机械拉伸的参与诱导AQP基因表达，我们在体内和体外实验设计。对于体内实验，带有球囊的小导管被沉淀到大鼠腹腔中，以扩大腹膜，而无需透析。对于体外实验，在橡胶板上培养了通过腹膜机械刮擦获得的间皮细胞，随后施加到30％的统计拉伸或60 Hz循环的拉伸。体内腹膜扩张在第3天产生了AQP基因表达的瞬时增强，这与实验性透析相似。但是，体外实验没有显着差异，这表明腹膜扩张的间接作用可能与AQP基因表达的瞬时增强有关。在临床上，在PD停用几周后，有时会经历CAPD患者超滤失败的意外恢复。腹膜膨胀引起的AQP基因表达可能与现象有关。较少的