Three main short-chain fatty acids attenuate 5-FU-induced THP-1 cells inflammation via glycerolphospholipid and sphingolipid metabolism

Abstract

Abstract 5-Fluorouracil (5-FU) is a common antitumor drug, but there is no effective treatment for its side effect, intestinal mucositis. The inflammatiory reaction of macrophages in intestinal mucosa induced by 5-FU is an important cause of intestinal mucositis. In this study, we investigated the anti-inflammatory effects of the three main short chain fatty acids (SCFAs), sodium acetate (NaAc), sodium propionate (NaPc), and sodium butyrate (NaB), on human mononuclear macrophage-derived THP-1 cells induced by 5-FU. The inhibition mechanism of macrophage inflammation was studied by untargeted metabolomics. The THP-1 cells were pre-incubated with 100 μmol/L of NaAc, NaPc, and NaB for 24 h, then treated with 2.5 mmol/L 5-FU for 24 h. The expressions of ROS, NF-κB p65, NLRP3 inflammasome, pro-inflammatory/anti-inflammatory cytokines were determined, and the cell metabolites was analyed by untargeted metabolomics techniques. It is found that the three main SCFAs could inhibite the pro-inflammation factors expressions including NLRP3, Caspase-1, IL-1β, and IL-6 when treated with 5-FU. The ROS expression and NF-κB pathway activity of THP-1 cell were inhibited by the three main SCFAs pre-incubated. Our results indicated that the three main SCFAs can effectively suppress the THP-1 cell inflammation via ROS/NF-κB/NLRP3 pathway, and affect 20 kinds of THP-1 cell metabolites which belong to amino acids and phosphatidylcholine. These significantly altered metabolites were involved in amino acid metabolism, glycerolphospholipid metabolism and sphingolipid metabolism., This is the first time that the three main SCFA had been found to inhibit 5-FU-induced macrophage inflammation through the above metabolic pathways through untargeted metabolomics.