Fig. 8. EUE drives Nrf2-dependent induction of HO-1 in BV-2 microglial cells. (A and C) EUE induces HO-1 expression in a time-dependent manner. Cells were treated with 100 μg/ml EUE for 0.5, 1, 2, 4, 6, 12, and 24 h. The protein and mRNA levels of HO-1 were measured by Western blot and RT-PCR analysis, respectively. (B and D) EUE induces HO-1 expression in a concentration-dependent manner. Cells were treated with the indicated concentrations of EUE for 6 or 12 h, and the protein and mRNA levels of HO-1 were determined by Western blot and RT-PCR analysis, respectively. (B) Western blot analysis of the expression levels of Nrf2, HO-1, β-actin, and GAPDH. Densitometric results are presented as means ± S.E.M. (n=3). (D) RT-PCR analysis of the mRNA levels of HO-1 and β-actin. Expression results are presented as means ± S.E.M. (n=3). *p<0.05; **p<0.01; and ***p<0.001 compared with the control group. (E and F) Treatment with ZnPP prevents EUE-mediated inhibition of LPS-stimulated production of NO (E) and PGE2 (F). Cells were pretreated with the indicated concentrations of ZnPP for 1 h, incubated with 100 μg/ml EUE for 30 min, and then stimulated with 100 ng/ml LPS for 24 h. The concentrations of nitrite in the culture medium were then determined using the Griess reagent. Data are presented as means ± S.E.M. (n=6). ***p<0.001 compared with the control group. ###p<0.001 compared with the LPS-treated group. #p<0.01 and ###p<0.001 compared with the EUE+LPS-treated group.
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