FIG. 6.
Lenti-ACE2 suppressed AngII-induced fibroblast migration and α-collagen I synthesis by inhibiting the NOX4/ROS/RhoA/Rock pathway. Primary lung fibroblasts were infected with lenti-ACE2 for 96 h and then exposed to AngII (10−7 M) for 24 h after pretreatment with A779 for 1 h. (A) Overexpression of ACE2 was confirmed with Western blot. (B, C) Active RhoA was determined using the GST pull-down assay; NOX4 and p-moesin protein were detected with Western blot. (D) TGFβ and Rock2 mRNA levels were measured with real-time RT-PCR. (E) The H2O2 concentration was measured. (F, G) Lung fibroblast migration assays. (H, I) Western blot was performed to determine the CTGF, α-SMA, and α-collagen I protein levels. Data are the means±SD of three independent experiments. Scale bar=200 μm. *p<0.05 versus control; †p<0.05 versus lentiNC; ‡p<0.05 versus AngII or lentiNC+AngII; §p<0.05 versus lentiACE2+AngII. ACE2, angiotensin-converting enzyme 2. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars