Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis.
Connective tissue growth factor (CTGF/CCN2) is one of the candidate factors mediating fibrogenic activity of TGF-beta. It was shown previously that the blockade of CTGF by antisense oligonucleotide (ODN) inhibits TGF-beta-induced production of fibronectin and type I collagen in cultured renal fibroblasts. The in vivo contribution of CTGF in renal interstitial fibrosis, however, remains to be clarified. With the use of a hydrodynamics-based gene transfer technique, the effects of CTGF antisense ODN are investigated in rat kidneys with unilateral ureteral obstruction (UUO). FITC-labeled ODN injection via the renal vein showed that the ODN was specifically introduced into the interstitium. At day 7 after UUO, the gene expression of CTGF, fibronectin, fibronectin ED-A, and alpha1(I) collagen in untreated or control ODN-treated obstructed kidneys was prominently upregulated. CTGF antisense ODN treatment, by contrast, markedly attenuated the induction of CTGF, fibronectin, fibronectin ED-A, and alpha1(I) collagen genes, whereas TGF-beta gene upregulation was not affected. The antisense treatment also reduced interstitial deposition of CTGF, fibronectin ED-A, and type I collagen and the interstitial fibrotic areas. The number of myofibroblasts determined by the expression of alpha-smooth muscle actin was significantly decreased as well. Proliferation of tubular and interstitial cells was not altered with the treatment. These findings indicate that CTGF expression in the interstitium plays a crucial role in the progression of interstitial fibrosis but not in the proliferation of tubular and interstitial cells during UUO. CTGF may become a potential therapeutic target against tubulointerstitial fibrosis.
Yokoi
Hideki
H
Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Mukoyama
Masashi
M
Nagae
Tetsuya
T
Mori
Kiyoshi
K
Suganami
Takayoshi
T
Sawai
Kazutomo
K
Yoshioka
Tetsuro
T
Koshikawa
Masao
M
Nishida
Takashi
T
Takigawa
Masaharu
M
Sugawara
Akira
A
Nakao
Kazuwa
K
D000199:Actins / Q000096:biosynthesis; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division; D055513:Connective Tissue Growth Factor; D005353:Fibronectins / Q000096:biosynthesis; D005355:Fibrosis / Q000378:metabolism*; D018014:Gene Transfer Techniques; D017874:Immediate-Early Proteins / Q000096:biosynthesis*; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis*; D007668:Kidney / Q000378:metabolism; D007674:Kidney Diseases / Q000378:metabolism*; D008297:Male; D008856:Microscopy, Fluorescence; D009130:Muscle, Smooth / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D010957:Plasmids / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D014162:Transfection; D015854:Up-Regulation; D014517:Ureteral Obstruction
D000199:Actins; C526316:CCN2 protein, rat; D005353:Fibronectins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D055513:Connective Tissue Growth Factor