Berberine Inhibition of Fibrogenesis in a Rat Model of Liver Fibrosis and in Hepatic Stellate Cells

Figure 2

BBR induced apoptosis in constitutively activated hHSC. (a) Constitutively activated hHSC in 96-well plate were treated with BBR for 24 hr and cytotoxicity of BBR was determined by MTT assay. The IC₅₀ of BBR on hHSC was roughly 75 μM. (b) hHSC were treated with BBR for 12 hr or 24 hr. Apoptosis and necrosis were analyzed by Annexin V/PI dual staining and flow cytometry. (c) BBR-treated hHSC were stained with JC-1 (10 μg/mL). Significant increase of green fluorescence with loss of red fluorescence indicated the relapse of mitochondrial membrane integrity. (d) BBR regulated Bcl-2/Bax ratio in hHSC. The cells were treated with BBR for 24 hr and cell lysates were subjected to Western blot hybridization. Increase of cleaved caspase-3 and caspase-9 and PARP indicated BBR induced apoptosis, while cyto C released from mitochondria exhibited loss of membrane integrity. Upregulation of Bax with Bcl-2 inhibition was observed in BBR-treated cells.

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