A NEW THERAPEUTIC STRATEGY FOR PERITONEAL SCLEROSIS ; USE OF ANTISENSE OLIGONUCLEOTIDES (ODN) AGAINST HSP47 IN RAT CHLORHEXIDINE GLUCONATE MODEL

腹膜硬化症是持续非卧床腹膜透析(CAPD)的严重并发症，以胶原堆积为特征。HSP47是一种胶原特异的分子伴侣，对胶原分子的生物合成和分泌是必不可少的。在先前的研究中，我们发现腹膜透析患者和大鼠腹膜硬化症模型的腹膜组织中热休克蛋白47的表达增加，本研究观察了反义寡核苷酸对腹膜硬化症进展的抑制作用。连续14天每日腹腔注射CG造成腹膜硬化症。从第8天开始给予反义或正义HSP47ODN和CG。第14天处死大鼠，取出腹膜组织。免疫组织化学方法检测HSP47的表达。同时检测III型胶原、α-平滑肌肌动蛋白和ED-1的表达。CG处理组腹膜增厚的腹膜间皮下区HSP47和III型胶原、α-平滑肌肌动蛋白的表达增加，而反义治疗组的表达和ED-1阳性细胞数明显低于CG组和正义处理组。CG组和正义组HSP47、α-平滑肌肌动蛋白和III型胶原的表达差异无统计学意义。我们的结果表明，HSP47在腹膜硬化症的胶原堆积过程中起着重要作用。提示用针对HSP47的反义寡核苷酸抑制HSP47可能是CAPD腹膜硬化症患者的一种新的治疗策略。

Peritoneal sclerosis, characterized by collagen accumulation, is a serious complication in continuous ambulatory peritoneal dialysis (CAPD). HSP47, a collagen-specific molecular chaperon, is essential for biosynthesis and secretion of collagen molecules. In the previous study, we demonstrated an increased expression of HSP47 in sclerotic peritoneum from CAPD patients and rat experimental model, induced by peritoneal injection of chlorhexidine gluconate (CG)s. In the present study, the inhibitory effects of antisense ODNs against HSP47 on the progression of peritoneal sclerosis was determined. Peritoneal sclerosis was developed by daily intra peritoneal injection of CG for 14 days. Antisense or sense HSP47 ODN was given together with CG from day 8. Rats were killed at day 14 and peritoneal tissue was taken out. We examined the expression of HSP47 by immunohistochemistry. The expression of collagen III, alpha-smooth muscle actin and ED-1 was also examined. The expression of HSP47 and collagen III, alpha-smooth muscle actin was increased in the thickened submesothelial areas of CG-treated peritoneal specimens, while those expression and the number of ED-1 positive cells were significantly reduced in antisense-treated group compared with those in CG and sense-treated groups. The expression of HSP47, alpha-smooth muscle actin and collagen III was not different between CG and sense-treated groups. Our results indicate that an important role of HSP47 in collagen accumulation during peritoneal sclerosis. It is suggested that inhibition of HSP47 by antisense ODNs against HSP47 may be a new therapeutic strategy in CAPD patients with peritoneal sclerosis.