Fig. 6

Effects of inhibiting (FK866 or 1MT) or sustaining (by NMN) NAD⁺ biosynthesis on bioenergetic profiles, inflammatory activation, and proliferation of HASMCs under high glucose conditions. A NAD⁺ levels detected by LC–MS. B Mitochondrial complex activity. C Mitochondrial stress test where different parameters of mitochondrial respiration are compartmentalized by a sequential application of Oligo, FCCP, and a combination of rotenone and antimycin. D Parameters of mitochondrial respiration. E Glycolysis stress test where different parameters of glycolytic flux are compartmentalized by a sequential application of glucose, Oligo, and 2DG. F Parameters of glycolytic flux. G Heatmap of mRNA expression of inflammatory cytokines determined by RT-qPCR. H Protein secretion of inflammatory cytokines. I Number of THP1 monocytes migrating toward HASMCs. J Proliferation of HASMCs. For all in vitro experiments, the control group is HG-cultured HASMCs. Data are presented as mean ± SD, n = 3 independent experiments. *P < 0.05; **P < 0.01. 1MT 1-methyl-L-tryptophan; NMN nicotinamide mononucleotide; Oligo oligomycin; R rotenone; A antimycin; OCR oxygen consumption rate; 2DG 2-deoxy-glucose; ECAR extracellular acidification rate; FC fold change; Gly glycolytic