3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), a food-born carcinogenic heterocyclic amine, promotes nitric oxide production in murine macrophages

A heterocyclic amine, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) is one of the potent food-born dietary carcinogens derived mainly from burnt meat products. In the present study, we investigated the inductive effect of Trp-P-1 on nitric oxide (NO) production in murine macrophages since NO and its oxidized derivatives are directly involved in triggering mutagenesis and carcinogenesis. Our results show that Trp-P-1 induced mRNA expression of inducible NO synthase (iNOS) and NO production without co-stimulation in murine peritoneal macrophages and RAW 264.7 cells. Trp-P-1 further enhanced both iNOS mRNA expression and NO production, which were primarily induced by lipopolysaccharide (LPS). Electrophoretic mobility shift assay demonstrated that Trp-P-1, alone or in the presence of LPS, facilitated the DNA binding activity of the transcription factor NF-κB, and the trans-acting activity of the NF-κB was confirmative as determined by in vitro transfection and a luciferase reporter gene assay. Moreover, Trp-P-1 induced increasing intracellular reactive oxygen species (ROS), which play an important role in NF-κB activation. These results suggest that Trp-P-1 induces NO production mediated by an increased intracellular ROS, NF-κB activation, and subsequent iNOS gene expression.