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Peritoneal membrane regeneration in peritoneal sclerosis

腹膜硬化中的腹膜再生

基本信息

批准号：
16590789
负责人：
MIYAZAKI Masanobu
金额：
$ 2.11万
依托单位：
Nagasaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590789/
关键词：
peritoneal dialysis peritoneal sclerosis HSP47 RNA interference mesothelial cells peritoneal regeneration

项目摘要

The main pathological features of peritoneal membrane from the patients with long PD therapy are accumulation of collagen and loss of mesothelial cells. These peritoneal alteration leads peritoneal membrane failure and is the biggest problem disturbing the longer-PD treatment more than 5-7 years. To maintain the longer PD treatment, peritoneal regeneration is an important issue. In the present study, we focused two points. One is inhibition of peritoneal sclerosis progression and other is regeneration of mesothelial cells. In the former, we tried to silence the gene of an essential molecular chaperon associated with collagen synthesis, heat shock protein 47 (HSP47). In order to inhibition of HSP47 expression, we used small interfering RNA (siRNA) combined with cationized gelatin microspheres (CGM) as vector. After three days of single injection of HSP47 siRNA with CGM to parietal peritoneum, experimental peritoneal sclerosis was made by chlorhexidine injection into peritoneal cavity. H … More SP47 siRNA significantly inhibited the progression of peritoneal sclerosis. Furthremore, biodegradable CGM containing HSP47 siRNA could continuously release siRNA over 21 days as a result of microsphere degradation. In the experiment of peritoneal regeneration, we examined the peritoneum after the fibrotic parietal peritoneum was stripped from abdominal wall in rats with chlorhexidine-induced experimental peritoneal sclerosis. Seven days after the stripping procedure, the peritoneum was covered with cells positive for cytokeratins and HBME-1, markers for mesothelial cells. Scanning electron microscopy also showed microvilli at the surface of the cells, which were main morphological feature of peritoneal mesothelial cells. Immunochemical analysis demonstrated the appearance of cells positive for vimentin, a marker of mesenchymal cells before the regeneration of mesothelial cells. In conclusion, HSP47 gene silencing by siRNA method combined with CGM was effective for the inhibition of progression of experimental peritoneal sclerosis and mesothelial cells had possibility of regeneration even on the fibrotic peritoneum. Less

长期腹膜透析患者腹膜的主要病理特征是胶原积聚和间皮细胞丢失。这些腹膜改变导致腹膜功能衰竭，是困扰5-7年以上长期腹膜透析治疗的最大问题。为了维持更长时间的腹膜透析治疗，腹膜再生是一个重要的问题。在本研究中，我们重点关注两点。一种是抑制腹膜硬化进展，另一种是间皮细胞再生。在前者中，我们试图沉默与胶原合成相关的重要分子伴侣热休克蛋白47（HSP 47）的基因。为了抑制HSP 47的表达，我们采用小干扰RNA（siRNA）结合阳离子化明胶微球（CGM）作为载体。将HSP 47 siRNA与CGM一起单次注射于腹膜壁层，连续3 d后，腹腔注射洗必泰造成实验性腹膜硬化。H ...更多信息 SP 47 siRNA显著抑制腹膜硬化的进展。此外，由于微球降解，含有HSP 47 siRNA的可生物降解CGM可持续释放siRNA超过21天。在腹膜再生实验中，我们观察了洗必泰诱导的实验性腹膜硬化大鼠的纤维化壁层腹膜从腹壁剥离后的腹膜。剥离程序后7天，腹膜覆盖有细胞角蛋白和HBME-1阳性的细胞，细胞角蛋白和HBME-1是间皮细胞的标志物。扫描电镜下可见细胞表面有微绒毛，这是腹膜间皮细胞的主要形态特征。免疫化学分析表明，波形蛋白阳性细胞的外观，间充质细胞再生前的间充质细胞的标记。结论：siRNA沉默HSP 47基因联合CGM可有效抑制实验性腹膜硬化的进展，即使在纤维化腹膜上，间皮细胞也有再生的可能。少