Aim: To investigate the enhancement of enhancers on matrine in vitro and in vivo. Methods: The permeability of matrine across rat intestine was determined by non-everted intestinal sacs model in vitro, and the in vivo activity was assayed in a model of acute liver injury in mice. Sodium deoxycholate (SDC) was selected as the enhancer Candidate. Results: Among the tested absorption enhancers, only polysorbate 80 (Tween 80) and SDC showed to be capable of increasing the transport of matrine through small intestinal mucosa in rats.Enhancement ratios of 1.6 was observed for 0.5% SDC solutions with the best absorption enhancement for matrine in vitro, it was chosen to conduct hepatoprotective study in vivo. In mice with acute liver injury, combination of matrine with 0.5% SDC demonstrated considerably more activity than matrine along after oral administration. Conclusion: The results indicated that the absorption of matrine could be significantly improved by the absorption enhancer.