Effect of salvianolic acid B on collagen production and mitogen-activated protein kinase activity in rat hepatic stellate cells.
OBJECTIVE
To investigate the mechanism of salvianolic acid-B (SA-B) action against liver fibrosis relating to mediating hepatic stellate cell (HSC) activation and transforming growth factor-beta1(TGF-beta1) intracellular signal transduction.


METHODS
HSC was isolated from normal rat through in situ perfusion of liver with pronase E and density-gradient centrifugation with 11 % nycondenz, then cells were subcultured. Cell proliferation was observed by [3H]TdR uptake. Cellular collagen deposition was measured with Ponceau S stain and semi-quantified with image analytic system. Type I collagen secretion in the supernatant was detected with ELISA. The gene expression of type I pro-collagen was analyzed by RT-PCR. The supernatant was acidified and active TGF-beta1 contents were assayed with ELISA. Mitogen-activated protein kinase (MAPK) activity was analyzed with immunoprecipitation and Western blot.


RESULTS
SA-B 0.1, 1, 10, and 100 micromol/L suppressed HSC proliferation concentration-dependently as determined by [3H]TdR uptake by 94.1 %, 82.4 %, 62.7 %, and 4 % of the control respectively (P<0.05 or P<0.01). SA-B 1, 10, and 100 micromol/L inhibited soluble type I collagen secretion by 75.3 %, 69.8 %, and 63.5 % of the control and decreased the matrix collagen deposition to 86.2 %, 75.4 %, and 73.4 % (P<0.05 or P<0.01). SA-B 1 and 10 micromol/L decreased the cell active TGF-beta1 secretion by 63.3 % and 15.6 % of the control, down-regulated pro-collgen alpha1(I) mRNA expression to 77.0 % and 51.8 % respectively (P<0.05). SA-B 1 and 10 micromol/L also inhibited MAPK activity by 1 to 2 fold respectively.


CONCLUSIONS
SA-B inhibited HSC proliferation and collagen production as well as decreased the cells' TGF-beta1 autocrine and MAPK activity, which might contribute to the mechanism of SA-B action against hepatic fibrosis.
Liu
Ping
P
Institute of Liver Disease, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Liuliver@online.sh.cn
Liu
Cheng-Hai
CH
Wang
Hai-Nan
HN
Hu
Yi-Yang
YY
Liu
Cheng
C
D000818:Animals; D001572:Benzofurans / Q000494:pharmacology*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053630:Depsides
D001572:Benzofurans; D024042:Collagen Type I; D020929:Mitogen-Activated Protein Kinase Kinases; D016212:Transforming Growth Factor beta; C076944:salvianolic acid B; D053630:Depsides