Antisense oligonucleotides against collagen-binding stress protein HSP47 suppress peritoneal fibrosis in rats.
BACKGROUND
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).


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Nishino
Tomoya
T
Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan.
Miyazaki
Masanobu
M
Abe
Katsushige
K
Furusu
Akira
A
Mishima
Yoko
Y
Harada
Takashi
T
Ozono
Yoshiyuki
Y
Koji
Takehiko
T
Kohno
Shigeru
S
D000199:Actins / Q000235:genetics; D000818:Animals; D018932:Chemokine CCL2 / Q000235:genetics; D003094:Collagen / Q000378:metabolism; D053331:Ectodysplasins; D005169:Factor VIII / Q000378:metabolism; D005355:Fibrosis; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D017403:In Situ Hybridization; D007274:Injections, Intraperitoneal; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D009130:Muscle, Smooth / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D010537:Peritoneum / Q000187:drug effects* / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016212:Transforming Growth Factor beta / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D000199:Actins; D018932:Chemokine CCL2; C506018:EDA protein, human; D053331:Ectodysplasins; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C494662:Serpinh1 protein, rat; D016212:Transforming Growth Factor beta; D042461:Vascular Endothelial Growth Factor A; D005169:Factor VIII; D003094:Collagen