Figure 6. β-hydroxybutyrate (HB) regulates insulin-induced inflammation. (A) Levels of p-p65 and total p65 noticeably diminished after treatment with 0.1–2 mM HB for 3 h, followed by incubation with or without 100 nM insulin for 2 h. (B) HEK293T cells were transiently transfected with a NF-κΒ and NF-κΒ-containing plasmid linked to the luciferase gene, pre-incubated with HB (0.25–1 mM) for 4 h, and then treated with insulin for 24 h. Results are presented in relative luminescence units (RLU). Results were obtained using one-factor ANOVA: $$$p<0.001 vs. pcDNA transduced cells; ##p<0.01 vs. NF-κB-luciferase transduced cells; *p<0.05 vs. insulin with NF-κB-luciferase transduced cells. (C) Confocal laser microscopy analysis of intracellular ROS levels using 2’,7’-dichlorodihydrofluorescein diacetate. Cells were incubated with 100 nM insulin for 2 h after pretreatment with HB for 3 h. Panel 1, normal; panel 2, 100 nM insulin; panel 3, 100 nM insulin + 0.5 mM HB.