Glycyrrhizin (Gly) and matrine (Mat) have been used for the treatment of patients with acute and chronic liver diseases including liver cirrhosis. We have previously reported that combined use of Gly and Mat (Gly+Mat) could protect liver function and prevent hepatocellular carcinoma (HCC) from occurring more effectively compared with Gly or Mat alone without causing significant adverse effects. However, whether Gly+Mat could produce a better therapeutic effect on liver fibrosis has not been investigated. In this study, we used a rat hepatic stellate cell line (HSC-T6) as an in vitro assay system to detect cell proliferation, collagen I, hexadecenoic acid (HA) and laminin (LN) secreted by HSCs. Serum HA, LN, procollgan type-III (PC-III) and pathologic changes were determined in a rat model of CCL₄-induced liver fibrosis. The results showed that Gly+Mat had a better effect in inhibiting the proliferation of activated HSC and diminishing collagen I and HA levels secreted by HSC as compared with Gly or Mat alone. Gly+Mat significantly reduced serum HA, LN and PC-III levels in the rat model of CCL₄-induced liver fibrosis compared with Gly or Mat alone. Hepatic histological analysis also confirmed that Gly+Mat administration exhibited the recovery effect remarkably, and could improve liver fibrosis in vitro and in vivo more effectively. The therapeutic strategy of Gly+Mat may prove to be a therapeutic option and preventive measure against hepatic fibrosis.