Nishino Tomoya, Miyazaki Masanobu, Abe Katsushige, Furusu Akira, Mishima Yoko, Harada Takashi, Ozono Yoshiyuki, Koji Takehiko, Kohno Shigeru
Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan.
Kidney Int. 2003 Sep;64(3):887-96. doi: 10.1046/j.1523-1755.2003.00169.x.
Peritoneal fibrosis is a serious complication in patients on continuous ambulatory peritoneal dialysis (CAPD), but the molecular mechanism of this process remains unclear. Heat shock protein 47 (HSP47), a collagen-specific molecular chaperone, is essential for biosynthesis and secretion of collagen molecules, and is expressed in the tissue of human peritoneal fibrosis. In the present study, we examined the effect of HSP47 antisense oligonucleotides (ODNs) on the development of experimental peritoneal fibrosis induced by daily intraperitoneal injections of chlorhexidine gluconate (CG).
HSP47 antisense or sense ODNs were injected simultaneously with CG from day 14, after injections of CG alone. Peritoneal tissue was dissected out 28 days after CG injection. The expression patterns of HSP47, type I and type III collagen, alpha-smooth muscle actin (alpha-SMA), as a marker of myofibroblasts, ED-1 (as a marker of macrophages), and factor VIII were examined by immunohistochemistry.
In rats treated with CG alone, the submesothelial collagenous compact zone was thickened, where the expression levels of HSP47, type I and type III collagen and alpha-SMA were increased. Marked macrophage infiltration was also noted and the number of vessels positively stained for factor VIII increased in the CG-treated group. Treatment with antisense ODNs, but not sense ODNs, abrogated CG-induced changes in the expression of HSP47, type I and III collagen, alpha-SMA, and the number of infiltrating macrophages and vessels.
Our results indicate the involvement of HSP47 in the progression of peritoneal fibrosis and that inhibition of HSP47 expression might merit further clinical investigation for the treatment of peritoneal fibrosis in CAPD patients.
腹膜纤维化是持续性非卧床腹膜透析（CAPD）患者的一种严重并发症，但其发生过程的分子机制仍不清楚。热休克蛋白47（HSP47）是一种胶原特异性分子伴侣，对胶原分子的生物合成和分泌至关重要，且在人类腹膜纤维化组织中表达。在本研究中，我们检测了HSP47反义寡核苷酸（ODNs）对每日腹腔注射葡萄糖酸氯己定（CG）诱导的实验性腹膜纤维化发展的影响。
从单独注射CG后的第14天起，将HSP47反义或正义ODNs与CG同时注射。CG注射28天后取出腹膜组织。通过免疫组织化学检测HSP47、I型和III型胶原、α平滑肌肌动蛋白（α-SMA，作为肌成纤维细胞的标志物）、ED-1（作为巨噬细胞的标志物）和因子VIII的表达模式。
在单独接受CG治疗的大鼠中，间皮下胶原致密区增厚，其中HSP47、I型和III型胶原以及α-SMA的表达水平升高。CG治疗组还观察到明显的巨噬细胞浸润，且因子VIII阳性染色的血管数量增加。反义ODNs治疗而非正义ODNs治疗消除了CG诱导的HSP47、I型和III型胶原、α-SMA表达的变化以及浸润巨噬细胞和血管的数量变化。
我们的结果表明HSP47参与腹膜纤维化的进展，抑制HSP47表达可能值得进一步临床研究用于治疗CAPD患者的腹膜纤维化。
