Fig. 9.

NMN ameliorated the inflammatory and oxidative responses in BV-2 cells stimulated with LPS for 24 h in vitro. (A) NAD+ in BV-2. (B) Supernatant TNF-α. (C) Supernatant IL-6. (D) Supernatant. IL-1β. (E, F) The ROS level of BV-2 as detected by flow cytometry. (G) The SOD activity of BV-2. (H) MDA content in BV-2. (I) Protein expression of SIRT1, PGC-1α, and acetylation level of lysine in BV-2. (J–K) mRNA expression of SIRT1, PGC-1α in BV-2. (L) Phosphorylation of P38 and P65. (M) CpG island of SIRT1 gene with location and sequence of methylation-specific primers. (N) Methylation status of cytosine residues in SIRT1 promoter. (O) BSP assay identified the hypermethylation status of SIRT1. Each circle represents a CG site, the black circles represent methylation and the white circles represent unmethylation. Each row represents a sequencing result of a TA clone in a sample. Tukey post-hoc test following one-way ANOVA was used. The values were presented as mean ± SD (*p < 0.05, **p < 0.01,***p < 0.001, n = 3–6 for each group).