Effects of osthole on LPS-induced MCP-1 secretion, ROS generation, and activation of NF-κB and Nrf2 in mesangial cells.

<p>(A) Mesangial cells (3×10⁴ in 1 ml of medium) were incubated for 2 h with or without osthole as indicated, then for 24 h with or without addition of 2 µg/ml LPS. MCP-1 concentration in conditional medium was measured by ELISA. (B) pNF-κB p65 activation was measured by an ELISA-based activity assay. (C) Mesangial cells (1×10⁶ in 1 ml of medium) were incubated for 30 min with or without 50 µM osthole or 10 mM NAC, then for 0–40 min with or without addition of 2 µg/ml LPS. ROS production was measured as the relative fluorescence intensity, as described in the Materials and Methods. (D) Representative Western blots for nuclear Nrf2 with lamin A as internal control. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.005.