NIM811, a nonimmunosuppressive cyclosporine analogue, suppresses collagen production and enhances collagenase activity in hepatic stellate cells.
OBJECTIVE
A recent decrease in patient survival has been reported among hepatitis C virus (HCV)-infected liver transplant recipients and this may be attributable to progression of fibrosis. We reported previously that cyclosporine suppressed the proliferation of, and collagen production in, hepatic stellate cells (HSCs). Here, we investigated the effects of NIM811, a cyclosporine analogue, on cell growth, collagen production and collagenase activity in HSCs.


METHODS
Rat HSCs and human HSC-derived TWNT-4 cells were cultured for the study. The expression of collagen, matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagenase activity was evaluated. Cell proliferation and apoptosis were measured. Phosphorylation of mitogen-activated protein kinases (MAPKs), Smad2 and Smad3 was evaluated. The expression of the tumour growth factor-beta (TGF-beta)-receptor and Smad7 genes was also evaluated.


RESULTS
NIM811, as well as cyclosporine, suppressed the transcription and synthesis of collagen and stimulated the production of MMP-1 with a concomitant enhancement of collagenase activity, although it did not change the expression of TIMP-1. NIM811 inhibited proliferation without induction of apoptosis. In the MAPKs and TGF-beta signalling pathways, NIM811 enhanced the phosphorylation of JNK and p38, but not extracellular signal-regulated kinases 1 and 2, and suppressed the phosphorylation of Smad2 and Smad3, accompanied by increased Smad7 transcription and decreased TGF-beta-receptor transcription.


CONCLUSIONS
These findings demonstrate that NIM811 not only suppresses collagen production and proliferation but also increases collagenase activity. These effects are accompanied by inhibition of TGF-beta signalling pathways.
Kohjima
Motoyuki
M
Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Enjoji
Munechika
M
Higuchi
Nobito
N
Kotoh
Kazuhiro
K
Kato
Masaki
M
Takayanagi
Ryoiichi
R
Nakamuta
Makoto
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D024042:Collagen Type I / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D017364:Collagenases / Q000378:metabolism*; D016572:Cyclosporine / Q000494:pharmacology*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008099:Liver / Q000166:cytology; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism
D024042:Collagen Type I; D051899:Smad2 Protein; C497784:Smad2 protein, rat; D051900:Smad3 Protein; D019715:Tissue Inhibitor of Metalloproteinase-1; D016572:Cyclosporine; C090438:(melle-4)cyclosporin; D017364:Collagenases; D020781:Matrix Metalloproteinase 1