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PD治疗患者的腹膜膜的主要病理特征是胶原蛋白的积累和间皮细胞的丧失。这些腹膜改变会导致腹膜膜破坏，并且是扰乱超过5 - 7年的较长PD治疗的最大问题。为了维持较长的PD治疗，腹膜再生是一个重要的问题。在本研究中，我们重点介绍了两点。一种是抑制腹膜硬化进展，另一种是间皮细胞的再生。在前者中，我们试图使与胶原蛋白合成，热休克蛋白47（HSP47）相关的基本分子链烯的基因保持沉默。为了抑制Hsp47表达，我们使用了与阳离子明胶微球（CGM）相结合的小干扰RNA（siRNA）作为载体。在将CGM单次注射Hsp47 siRNA至顶叶腹膜的三天后，实验性腹膜硬化H…更多的SP47 siRNA显着抑制腹膜硬化的进展。此外，由于微球降解，含有HSP47 siRNA的可生物降解的CGM可以在21天内继续释放siRNA。在腹膜再生的实验中，我们检查了纤维化顶叶腹膜腹膜腹膜的腹膜腹膜，从腹壁中剥离了用氯己定引起的实验性腹膜硬化。剥离手术后的七天，腹膜被细胞角蛋白和HBME-1阳性的细胞覆盖，这是间皮细胞的标记。扫描电子显微镜还显示了细胞表面的微绒毛，这是腹膜间皮细胞的主要形态特征。免疫化学分析表明，在间质细胞再生之前，波形蛋白的细胞呈阳性。总之，通过siRNA方法与CGM结合使用的Hsp47基因沉默对于抑制实验性腹膜硬化的进展和间皮细胞的进展也有可能再生于纤维性腹膜。较少的