Abstract: OBJECTIVE To study the effects of Liangxue Huayu Fang on pathways related to autophagy and apoptosis of hepatocyte injury induced by tumor necrosis factor alpha (TNF-α) combined with D- galactosamine(D-GalN)， and to explore the molecular regulative mechanism.METHODS The normal human liver cells LO2 were cultured in vitro. The hepatocyte injury model was induced by TNF- α (100 ng/mL) and D-GalN(44 μg/mL). Fluorescence microscope was used to observe the morphological changes of cells. The cell viability was detected by MTT; Western blot was adopted to detect the apoptosis and autophagy marker， such as proteins caspase-3， Beclin1， LC3 Ⅰ， LC3 Ⅱ; changes of mTOR protein expression level of apoptosis and autophagy regulating protein Bax， Bcl-2 and mTOR signal pathway were also detected.RESULTS Liangxue Huayu Fang can improve pathological injury of hepatocyte with a dose-dependence; cell apoptosis was decreased and the expression of autophagy markers， such as protein caspase-3， Beclin1， LC3 Ⅱof the treatment group showed decreased and the expression of negative correlative Bcl-2 protein was up-regulated while the positive correlative Bax expression was down-regulated. All the data showed a dose - response relationship to some degree and the expression of mTOR protein showed down-regulated with dose - response.CONCLUSION The protective effects of Liangxue Huayu Fang on hepatocyte injury induced by TNF-α and D-GalN may be related to inhibition of autophagy and apoptosis. Its main mechanism may relate to regulation of the expression of proteins participating in autophagy and apoptosis.