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

腹膜硬化症的新治疗策略；

• 批准号: 11671039
• 负责人: OZONO Yoshiyuki
• 金额: $ 1.15万
• 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671039/
• 关键词: CAPD; HSP47; peritoneal tissue; antisense ODN for HSP 47; rat peritonear sclerosis model; HSP 47; HSP47; 腹膜硬化; 線維化

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.

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。

项目成果

Shioshita K: "Expression of heat shock proteins 47 and 70 in the peritoneum of patients on continuous ambulatory peritoneal dialysis"Kidney Int. 57.. 619-631 (2000)

Shioshita K：“连续非卧床腹膜透析患者腹膜中热休克蛋白 47 和 70 的表达”Kidney Int。

Miyazaki M: "HSP 47 in peritoneal sclerosis."Peritoneal dialysis 2000 (JIN TO TOSEKI). 49. 28-32 (2000)

Miyazaki M：“腹膜硬化中的HSP 47。”腹膜透析2000（JIN TO TOSEKI）。

潮下敬: "培養ヒト腹膜中皮細胞におけるHeat shock protein 47(HSP47)の発現の検討について."腹膜透析'99(腎と透析別冊47). 240-243 (1999)

Takashi Shioshita：“培养的人腹膜间皮细胞中热休克蛋白 47 (HSP47) 表达的检查。”腹膜透析 99（肾脏和透析特刊 47）（1999 年）。

潮下敬、宮崎正信、芦洋府美子、三島陽子、三宅千恵、大園恵幸、原田孝司、田口尚、河野茂: "培養ひと腹膜中皮細胞におけるHeat Shock Pnteir47(HSP47)の発現の検討について"腹膜透析99'(腎と透析別冊). 47. 240-243 (1999)

Takashi Shioshita、Masanobu Miyazaki、Fumiko Ashi、Yoko Mishima、Chie Miyake、Keiyuki Ozono、Takashi Harada、Hisashi Taguchi、Shigeru Kono：“检查培养的人腹膜间皮细胞中热休克 Pnteir47 (HSP47) 的表达”“腹膜透析 99” ”（肾脏和透析分册）47. 240-243 (1999)。

Abe K: "Interstitial expression of heat shock protein 47 and α-smooth muscle actin in renal allograft failure."Nephrol Dial Transplant. 15.. 529-535 (2000)

Abe K：“同种异体肾移植失败时热休克蛋白 47 和 α-平滑肌肌动蛋白的间质表达。”Nephrol Dial Transplant，15.. 529-535 (2000)