Figure 5

The DNA of Apoptotic hepatocyte up-regulates α-SMA expression in rat HSCs and FZHY inhibited hepatocyte apoptosis and the expressions of α-SMA and type I collagen in HSCs. (A) Fragmented apoptotic DNA in cultured hepatocytes was induced by 200 ng/ml Act D and 20 ng/ml TNF-α. Lane 1, Marker, Lane 2; phosphate-buffered saline (PBS)-treated hepatocytes; Lane 3, Act D/TNF-α treated hepatocytes. (B) Primary hepatocytes were incubated with or without 200 ng/ml ActD and 20 ng/ml TNF-α for 6 h. 100 μg/ml DNA from hepatocytes incubated with PBS acted as a control. 25 ~ 100 μg/ml DNA from apoptotic hepatocyte stimulated α-SMA expression in rat HSCs in a dose dependent manner (Immunofluorescence staining, × 400). (C) Western blot analysis of expressions of α-SMA and type I collagen in rat HSCs. (D) Graphic presentation of the relative expressions of α-SMA and type I collagen. The values were represented as the density of α-SMA and collagen I vs. housekeeping gene GAPDH (%) from 3 samples. (E) Primary hepatocytes were incubated with or without 200 ng/ml ActD and 20 ng/ml TNF-α for 6 h. Then, 100 μg/ml DNA from apoptotic hepatocytes was added into HSCs after culture for 4 days. 100 μg/ml DNA from hepatocytes incubated with PBS acted as a control. The apoptotic DNA from hepatocytes treated by FZHY or Z-VAD-FMK (FMK) had less stimulating effects on α-SMA expression in HSCs (Immunofluorescence staining, × 400). ^** P < 0.01 vs. Control; ^## P < 0.01 vscpc Act D/TNF-α group.