Effects of CCl₄ on VEGF expression in hepatocytes and effects of conditioned medium collected from hepatocytes on the activation and proliferation of hepatic stellate cells.

<p>(A)BRL cells were exposed to CCl₄ for 12 hours and then the culture medium was replaced with fresh DMEM. After another 24 hours of culture, the conditioned medium was collected. Real-time PCR and ELISA assays were used to assess VEGF expression in hepatocytes after exposure to CCl₄. (B) BRL cells were exposed to CCl₄ for 12 hours and then the culture medium was replaced with fresh DMEM. After another 24 hours of culture, the conditioned medium was collected. The primary HSCs were plated in 6-well plates (1×10⁶ cells/well) and treated by conditioned medium with or without bevacizumab (100 µg/ml) for 72 hours. Then the cells were harvested and real-time PCR was performed to assess the expression of α-SMA and TGF-β1 in HSCs. (C) CCK-8 assay was used to assess the effects of conditioned medium with or without bevacizumab (100 µg/ml) on the proliferation of the HSC-T6 cell line. (D) MTT assay was employed to examine the effects of conditioned medium with or without bevacizumab (100 µg/ml) on the proliferation of the HSC-T6 cell line. (*<i>P<0.05</i>).