Research Paper Volume 13, Issue 17 pp 21470—21482

SCFAs promote intestinal double-negative T cells to regulate the inflammatory response mediated by NLRP3 inflammasome

SCFAs promoted the formation of DNTs through OX40. (A–B) SCFAs promoted the formation of DNTs in vitro (x¯ ± s, n = 3): Cells in the SCFAs group were co-cultured with acetic acid (5 mM), propionic acid (1 mM) and butyric acid (1 mM) for 24 h, The proportion of DNTs in the SCFAs group was increased, and SCFAs promoted the formation of DNTs. Significant difference between groups, P C–D) Effect of OX40 inhibition on the formation of DNTs (x¯ ± s, n = 3): after OX40 inhibition, the proportion of DNTs in the SCFAs-anti-OX40 group was significantly down-regulated than that in the SCFAs group. Significant difference between the groups, P E) Detection of Fas and TNF-α secretion in DNTs (x¯ ± s, n = 3): the expression of Fas and TNF-α in DNTs was significantly up-regulated, which was significantly higher than that of the Con group. Comparison between groups, P F–G) Effect of SCFAs on the activation of OX40 in DNTs (x¯ ± s, n = 3): the expression of OX40, Bcl-2, Bcl-xl and Survivin was significantly up-regulated in the SCFAs group, which was higher than that of the Con group. Comparison with the Con group, *P H–I) Effect of OX40 inhibition on the expression of related protein (x¯ ± s, n = 3): anti-OX40 pretreatment could significantly inhibit the expression of Bcl-2, Bcl-xl and Survivin. Comparison with the Con group, *P #P

Figure 5. SCFAs promoted the formation of DNTs through OX40. (AB) SCFAs promoted the formation of DNTs in vitro (x¯ ± s, n = 3): Cells in the SCFAs group were co-cultured with acetic acid (5 mM), propionic acid (1 mM) and butyric acid (1 mM) for 24 h, The proportion of DNTs in the SCFAs group was increased, and SCFAs promoted the formation of DNTs. Significant difference between groups, P < 0.05. (CD) Effect of OX40 inhibition on the formation of DNTs (x¯ ± s, n = 3): after OX40 inhibition, the proportion of DNTs in the SCFAs-anti-OX40 group was significantly down-regulated than that in the SCFAs group. Significant difference between the groups, P < 0.05. (E) Detection of Fas and TNF-α secretion in DNTs (x¯ ± s, n = 3): the expression of Fas and TNF-α in DNTs was significantly up-regulated, which was significantly higher than that of the Con group. Comparison between groups, P < 0.05. (FG) Effect of SCFAs on the activation of OX40 in DNTs (x¯ ± s, n = 3): the expression of OX40, Bcl-2, Bcl-xl and Survivin was significantly up-regulated in the SCFAs group, which was higher than that of the Con group. Comparison with the Con group, *P < 0.05. (HI) Effect of OX40 inhibition on the expression of related protein (x¯ ± s, n = 3): anti-OX40 pretreatment could significantly inhibit the expression of Bcl-2, Bcl-xl and Survivin. Comparison with the Con group, *P < 0.05; comparison with the SCFAs group, #P < 0.05.