A novel Keap1 inhibitor iKeap1 activates Nrf2 signaling and ameliorates hydrogen peroxide-induced oxidative injury and apoptosis in osteoblasts

摘要

The primary murine osteoblasts (A–G) or the primary human osteoblasts (H–J) were treated with applied concentration of iKeap1 (0.1–10 μM), cells were further cultured for indicated time points, the relative ARE activity, NQO1 activity, and cell viability (CCK-8 OD) were tested (A); Keap1-Nrf2 association was tested by the co-immunoprecipitation (Co-IP) assays (B, H); Expression of listed proteins in cytosol fraction lysates and nuclear fraction lysates were examined by western blotting assays (C, E, G, I), with relative expression of listed mRNAs tested by qRT-PCR assays (F, J). Expressions of the listed proteins were quantified and normalized to the loading control. Quantified values were mean ± standard deviation (SD, n = 5). “C” stands for the untreated control cells. “Veh” stands for the vehicle control (0.1% DMSO). * P < 0.05 vs. “Veh” cells. Experiments were repeated five times, with similar results obtained.