Objective: Preparing rabbit serum containing Clastrus orbiculatus extracts (COE)

to investigate preliminarily the effects on proliferation and vascular endothelial growth factor-c (VEGF-c) expression in hepatoma cells of mice (Hepa1-6). Method: The New Zealand white rabbits received COE with dose of 20 mg ·kg-1 ·d-1 by gavage.The medicated rabbit serum was obtained using serum phamacological approach.Hepa1-6 cells were cultured in media with different dosages of COE-containing serum (the low

medium

high dose groups containing 10%

20%

30% medicated serum respectively).At the same time

the negative control group and 5-fluorouracil (5-Fu) positive control group were set up.The proliferation inhibition effect of medieated serum containing COE on Hepa1-6 cells was detected by microculture tetrazolium (MTT) assay and the expression level of VEGF-c of Hepa1-6 cells was tested by enzyme linked immunosorbent (ELISA). Result: The typical morphological changes of apoptosis were observed after incubation with COE-containing serum of each dose.The medium and high dose COE-containing serum could significantly inhibit the proliferation of Hepa1-6 cells time-dependently.The VEGF-c expression level of Hepa1-6 cells treating with medium and high dose COE-containing serum for 24 hours were significantly decreased compared with that of the negative control group (P<0.05). Conclusion: COE-containing serum can significantly inhibit proliferation and VEGF-c expression of Hepa1-6 cells in vitro.