Sulforaphane Suppresses Hepatitis C Virus Replication by Up-Regulating Heme Oxygenase-1 Expression through PI3K/Nrf2 Pathway
Fig 3
SFN inhibited HCV replication through HO-1 dependent pathway.
(A, B) Ava5 cells were treated with increasing concentrations of SnPP (0–20 μM) with or without 7.5 μM SFN for 3 days. HCV protein synthesis and RNA replication were analyzed by Western blotting and qRT-PCR, respectively. (C and D) Increasing amounts of HO-1-specific shRNA (0.25–2 μg) or nonspecific shRNA were transfected into Ava5 cells that were then treated with or without SFN (7.5 μM). After 3 days, HCV protein synthesis and RNA replication were analyzed by Western blotting and qRT-PCR, respectively. Western blotting was performed using antibodies against HCV NS5B and HO-1. An antibody against GAPDH was used to show equal loading of lysates. Relative RNA levels were normalized to the internal control, gapdh. The relative HCV RNA levels were presented as percentage changes compared to SFN-untreated/untranfected Ava5 cells, in which level was considered as 100%. Data were represented as the means of normalized data ± standard deviations (error bars) from five independent experiments. *P < 0.05; ** P < 0.01.