S-Propargyl-cysteine Exerts a Novel Protective Effect on Methionine and Choline Deficient Diet-Induced Fatty Liver via Akt/Nrf2/HO-1 Pathway

Figure 9

SPRC upregulated HO-1 expression via activation of Akt/Nrf2 pathway in HepG2 cells. The cells were incubated with SPRC (50 μM) for indicated periods (a) or indicated concentrations of SPRC for 24 h (b), and the expression of HO-1 and Nrf2 was detected by Western blot, and GAPDH was used as a loading control; data shown are means ± SEM, compared with untreated cells; data were from at least three independent experiments, each performed in duplicate. (c) The cells were pretreated with LY294002 or PAG for 30 min, then incubated with SPRC (50 μM) for 6 h, and then stimulated with or without OA (1.5 mM) for 18 h, and the expression of HO-1 and Nrf2 was detected by Western blot, and GAPDH was used as loading control; data shown are means ± SEM; compared with untreated cells; compared with OA-stimulated cells; data were from at least three independent experiments, each performed in duplicate. (d) The cells were treated with SPRC (50 μM) for indicated periods (0, 15, 30, 60, and 120 min), and the translocation of Nrf2 from cytosol to nucleus was detected by Western blot, and Lamin B1 was used as loading nuclear control and GAPDH was used as loading cytosol control; data shown are means ± SEM; compared with nuclear Nrf2 in untreated cells; compared with cytosol Nrf2 in untreated cells; data were from at least three independent experiments, each performed in duplicate. (e) The cells were pretreated with LY294002 and PAG for 30 min and then incubated with SPRC (50 μM) for 60 min, and the translocation of Nrf2 from cytosol to nucleus was detected by Western blot, and Lamin B1 was used as loading nuclear control and GAPDH was used as loading cytosol control; data shown are means ± SEM; compared with nuclear Nrf2 in untreated cells; compared with nuclear Nrf2 in SPRC-treated cells; compared with cytosol Nrf2 in untreated cells; compared with cytosol Nrf2 in SPRC-treated cells; data were from at least three independent experiments, each performed in duplicate.

(a)

(b)

(c)

(d)

(e)