Figure 10.

Obovatol enhanced the peroxidase activity of Prx2 and suppressed intracellular ROS production. Ferrous oxidation assay was carried out in 100 µL of reaction mixture containing DTT (2 mM), H₂O₂ (50 µM), obovatol (10 µM) and recombinant Prx2 protein. After incubation of the mixture for 10 min, the remaining H₂O₂ was measured based on the color changes after adding Fe(NH₄)SO₄ and potassium thiocyanate. DMSO and BSA were used as a vehicle control for obovatol and a negative control for Prx2 respectively. The data were expressed as the mean ± SD (n= 3). #P < 0.05, ##P < 0.01 versus H₂O₂ only; *P < 0.01 versus BSA+DMSO or Prx2+DMSO (A). The direct H₂O₂-scavenging effect of obovatol was also evaluated by ferrous oxidation assay in the absence of recombinant Prx2 protein (B). BV-2 microglia cells were treated with 100 ng·mL⁻¹ LPS for 30 min in the absence or presence of obovatol (10 µM), followed by 30 min incubation with 10 µM H₂DCF-DA. Fluorescence was measured by flow cytometer. The results were expressed as the mean fluorescence intensity (C). DMSO, dimethyl sulfoxide; Prx2, peroxiredoxin 2; ROS, reactive oxygen species; DTT, dithiothreitol; BSA, bovine serum albumin.