Somatostatin at nanomolar concentration reduces collagen I and III synthesis by, but not proliferation of activated rat hepatic stellate cells.
Previous studies have shown antifibrotic effects of somatostatin. Since hepatic stellate cells (HSC) express somatostatin receptors and play a key role in hepatic fibrogenesis, we investigated the in vitro antifibrotic effect of somatostatin on rat HSC. At day 12 after isolation, cells were exposed to different concentrations of somatostatin (10(-6)-10(-9) mol l(-1)). mRNA expression of collagen types I and III, and of smooth muscle alpha-actin (alpha-SMA) was analysed by Northern blotting. At 10(-9) mol l(-1), somatostatin significantly reduced mRNA expression of collagen I (72.3 +/- 10.7%; 95% confidence interval (95% CI): 45.5-99.0), collagen III (79.0 +/- 4.5%; 95% CI: 67.6-90.4) and alpha-SMA (65.7 +/- 5.9%; 95% CI: 51.1-80.2), as compared to control normalized at 100%. These results were confirmed by quantitative RT-PCR. Cycloheximide experiments indicated that somatostatin has no direct transcriptional effect.Using immunoprecipitation, we demonstrated that somatostatin also decreased de novo synthesis of collagen I (73 +/-10%; 95% CI: 48-98%), collagen III (65 +/- 13%; 95% CI: 33-97%) and alpha-SMA (47 +/- 9%; 95% CI: 25-69%). Remarkably, at higher concentrations, somatostatin did not suppress collagen mRNA expression nor de novo protein synthesis. We ascribe this observation to desensitization of the cells for somatostatin. Cell proliferation, as measured by 5-bromo-2'-deoxyuridine labelling, was not altered by somatostatin. No significant effect on the intermediate and actin cytoskeleton were detected by immunohistochemistry and Western blotting. Our findings imply that in vivo antifibrotic effects of somatostatin could result partially from a direct action of somatostatin on HSC, but other, in vivo effects are probably also involved.
Reynaert
Hendrik
H
Laboratory for Liver Cell Biology, Vrije Universiteit Brussel (VUB), Belgium.
Rombouts
Krista
K
Jia
Yutao
Y
Urbain
Daniel
D
Chatterjee
Nirjhar
N
Uyama
Naoki
N
Geerts
Albert
A
D000199:Actins / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013004:Somatostatin / Q000494:pharmacology*
D000199:Actins; D024042:Collagen Type I; D024061:Collagen Type III; D003513:Cycloheximide; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D013004:Somatostatin; C498862:Acta2 protein, rat