INHIBITORY EFFECTS OF THE HERBAL MEDICINE SHO-SAIKO-TO (TJ-9) ON CELL-PROLIFERATION AND PROCOLLAGEN GENE EXPRESSIONS IN CULTURED RAT HEPATIC STELLATE CELLS

Background/Aims: It is of extreme importance to prevent liver fibrosis and subsequent progression to liver cirrhosis. The aim of our study w as to elucidate in vitro whether Sho-saiko-to exerted inhibitory effec ts on hepatic stellate cells. Methods: Hepatic stellate cells were iso lated from male Wistar rats. Water-soluble ingredients of Shosaiko-to were obtained at concentrations of 10, 100, 250, 500 and 1000 mu g/ml. Morphological transformation was observed under a phase-contrast micr oscope. Flow cytometric analysis was performed on day 4 after culture to evaluate the potential to proliferate of the stellate cells by anal yzing cell cycles. Northern blot analysis was carried out on day 3 aft er culture to determine the expressions of type I and type III procoll agen mRNAs. Results: (i) Sho-saiko-to 500 and 1000 mu g/ml inhibited m orphological transformation of the stellate cells to myofibroblast-lik e cells. (ii) Sho-saiko-to 500 and 1000 mu g/ml significant ly (p<0.00 01) accumulated the cells in the G(0)/G(1) phase (118.8+/-0.7%, 119.2/-0.5%, respectively as compared with control) and significantly (p<0. 0001) decreased cell numbers subsequently in G(2)/M phase (47.5+/-8.1% ? 48.9+/-2.0%, respectively). (iii) Sho-saiko-to 500 and 1000 mu g/ml also significantly (p<0.05 or p<0.0001) suppressed procollagen mRNA ex pression of type I to 51.5+/-6.4%, 34.9+/-3.7%, respectively; and type III to 51.3+/-12.3%, 46.7+/-11.4%, respectively. Conclusions: We have clarified the inhibitory effects of Sho-saiko-to on hepatic stellate cells in vitro. Shosaiko-to could be a potent inhibitor in the pathoge nesis of liver fibrosis.