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Abstract Objective: To study the effects of three main short-chain fatty acids (SCFAs), i.e. sodium acetate, sodium propionate, and sodium butyrate, on 5-fluorouracilinduced inflammatory response in human mononuclear macrophage THP-1 cells and its potential mechanism. Methods: The THP-1 cells was divided into 5 groups: control group,THP-1 cells were not treated; 5-FU group, THP-1 cells were treated with 2.5 mmol/L 5-FU alone for 24 h; 5-FU+sodium acetate group, 5-FU+sodium propionate group and 5-FU+sodium butyrate group, all were pretreated with corresponding 100 μmol/L SCFAs for 24 h, and then 2.5 mmol/L 5-FU were added for 24 h. The qRT-PCR was used to detect the mRNA expression of various inflammation-related factors in cells. ELISA was used to detect the concentrations of IL-1β, IL-6, and IL-10 in the cell supernatant. Western blotting was used to detect the expression of TLR9, NLRP3, Caspase-1, LC3-Ⅱ, Beclin-1, and the expression of NF-κB p65 protein in the nucleus and cytoplasm. Flow cytometry was used to detect cellular reactive oxygen species. Results: Compared with the control group, TLR9, NLRP3, Caspase-1, IL-1β, IL-6, LC3-Ⅱ, Beclin-1, nuclear NF-κB p65 protein expression and reactive oxygen species in 5-FU group were increased significantly(P<0.01). Compared with 5-FU group, the expression of TLR9 in 5-FU+sodium acetate group was greatly reduced(P<0.01); the expression of IL-1β and reactive oxygen species in 5-FU+sodium propionate group were markedly reduced (P<0.01). the expression of IL-1β and nuclear NF-κB p65 protein in 5-FU+sodium butyrate group was greatly reduced (P<0.01), and intracytoplasmic expression was remarkably increased(P<0.01); three kinds of short-chain fatty acid pretreatments could significantly reduce NLRP3, Caspase-1, IL-6, LC3-Ⅱ and Beclin-1 expression(P<0.01), and increase the expression of anti-inflammatory factor IL-10 (P<0.01). Conclusion: Three short-chain fatty acids, sodium acetate, sodium propionate and sodium butyrate, could inhibit the inflammatory response of THP-1 cells induced by 5-FU, inhibit the level of autophagy, and reduce the amount of cellular reactive oxygen generation, which is possible by inhibiting the TLR9/NF-κB/ROS pathway.
［Key words］short-chain fatty acids; 5-fluorouracil; inflammatory reaction; autophagy; reactive oxygen species(ROS); NF-κB
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