Objective To investigate the anti-liver fibrosis mechanism of spleen-invigorating and blood-activating traditional Chinese medicine (TCM). Methods LX-2 cells were cultured in vitro, and CCK-8 assay was used to determine the optimal concentration of lyophilized powder of spleen-invigorating and blood-activating TCM in promoting the proliferation of LX-2 cells. The cells were divided into control group, model group ［induced by 5 ng/mL transforming growth factor-β1 (TGF-β1)］, and TCM group (5 ng/mL TGF-β1+lyophilized powder of spleen-invigorating and blood-activating TCM). Western blot and immunofluorescence assay were used to measure the expression levels of type Ⅰ collagen (Col-Ⅰ), type Ⅲ collagen (Col-Ⅲ), and α-smooth actin (α-SMA). Results CCK-8 assay showed that LX-2 cells induced by 1.5 mg/mL lyophilized powder of spleen-invigorating and blood-activating TCM had the highest cell proliferation rate (P<0.05). Western blot and immunofluorescence assay showed that compared with the control group, the model group had significant increases in the protein expression levels of Col-Ⅰ, Col-Ⅲ, and α-SMA (P<0.05), and compared with the model group, the TCM group had significant reductions in the expression levels of Col-Ⅰ, Col-Ⅲ, and α-SMA (P<0.05). Conclusion The spleen-invigorating and blood-activating TCM can improve the fibrosis of LX-2 cells by inhibiting the expression of Col-Ⅰ, Col-Ⅲ, and α-SMA.