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

腹膜硬化症是持续性非卧床腹膜透析（CAPD）的严重并发症，其特征是胶原积聚。HSP 47是一种胶原蛋白特异性分子伴侣，在胶原蛋白的合成和分泌过程中起重要作用。在前期的研究中，我们证实了腹膜内注射葡萄糖酸氯己定（CG）诱导CAPD患者和大鼠实验模型腹膜内HSP 47表达增加。在本研究中，针对HSP 47的反义ODNs对腹膜硬化的进展的抑制作用被确定。每天腹腔注射CG 14天，形成腹膜硬化。从第8天开始，将反义或正义HSP 47 ODN与CG一起给予。在第14天处死大鼠，取出腹膜组织。免疫组化法检测HSP 47的表达。还检测了III型胶原、α-平滑肌肌动蛋白和艾德-1的表达。与正常对照组和正常对照组相比，CG处理组大鼠腹膜间皮下增厚区HSP 47、Ⅲ型胶原、α-平滑肌肌动蛋白表达增加，而反义处理组HSP 47、Ⅲ型胶原、α-平滑肌肌动蛋白表达及艾德-1阳性细胞数减少。热休克蛋白47、α-平滑肌肌动蛋白和Ⅲ型胶原的表达在CG组和sense组之间没有差异。我们的研究结果表明，在腹膜硬化过程中，HSP 47在胶原积累中起重要作用。结论：HSP 47反义寡核苷酸抑制HSP 47的表达可能是治疗腹膜硬化CAPD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.