Fig. 7.

Ang_1–7 inhibits ERK phosphorylation induced by AngII in HSC. mRNA was isolated from hepatocytes (Hep), KCs, ECs, and HSCs and expression of ace2 and Mas was evaluated by qPCR (A,B). RT-PCR for Mas was performed using mRNA isolated from whole liver from balb/c mice subjected to BDL for 18 days (C). Mouse HSCs were cultured for 5 days and then serum starved for 24 hours. ROS production was evaluated by DCFD fluorescence of mouse HSCs stimulated with AngII (black solid line) and preincubated with Ang_1–7 (gray solid line) (D). Cells were stimulated with AngII (10⁻⁷ M) in the presence or absence of Ang_1–7 (10⁻⁷ M) for 15 minutes. Activation of signaling pathways was analyzed by immunoblotting for IκBα and using phospho-specific antibodies for p38, Akt, JNK, c-Jun, and ERK. Equal loading was evaluated by Ponceau S staining (not shown). Data are representative of three independent experiments.