Succinate-treated macrophages attenuate dextran sodium sulfate colitis in mice

Abstract

The safety and effectiveness of adalimumab was demonstrated in a phase 3 trial in Japanese patients with intestinal Behçet's disease. The aim of this study was to evaluate the long-term safety and effectiveness of adalimumab in Japanese patients with intestinal Behçet's disease.

Fig. 1.

Peritoneal cavity cell transfer in a dextran sulfate sodium (DSS)-colitis model. (A) Experimental design of the peritoneal cavity cells (PCCs) transfer. Donor mice were intraperitoneally injected with 40 mM succinate or not for 3 days. Recipient mice were transferred with the PCCs on day 0, were then administrated 2% DSS in drinking water until day 6 followed by normal water for 2 days and were sacrificed on day 8. (B, C) Gating strategy of peritoneal macrophage sorting. F4/80⁺ PCCs isolation using BD FACS Aria II. (B) Unstained PCCs. (C) Stained pre-sorting PCCs and post-sorting PCCs. HBSS, Hanks’ balanced salt solution; Veh, vehicle; Suc, succinate; SSC-A, side-scatter area; FSC-A, forward-scatter area; FSC-W, forward-scatter width; FSC-H, forward-scatter height; SSC-W, side-scatter width; SSC-H, sidescatter height; WT, water.

Fig. 2.

Transferred succinate-treated peritoneal cavity cells (Suc-PCCs) ameliorate dextran sulfate sodium (DSS)-induced colitis. The recipients of Suc-PCCs show the attenuated body weight change (A), disease activity index (DAI) (B), and colon length (C) compared to vehicle-treated PCC (Veh-PCC). The results were examined using Prism 5.0 software (GraphPad Inc.). Statistical significance was evaluated using one-way analysis of variance and P<0.05 were regarded to be significant. Bars in graphs represent mean±standard error of the mean. ^aP<0.05, ^bP<0.01, and ^cP<0.001 compared with the water group. ^dP<0.05, ^eP<0.01, and ^fP<0.001 compared with the Suc-PCC group. HBSS, Hanks’ balanced salt solution.

Fig. 3.

Periodic acid Schiff staining and quantification of colitis. (A) Section from colon tissues stained with periodic acid Schiff. Histopathologic scores (B) and quantification with integrated density (IntDen) of goblet cells (C) of colon tissues from mice were assessed by Image J v.1.496 software (National Institutes of Health). The results were examined using Prism 5.0 software (GraphPad Inc.). Statistical significance was evaluated using one-way analysis of variance and t-test, respectively, P<0.05 were considered significant. Bars in graphs represent mean±standard error of the mean. ^aP<0.05, ^bP<0.001 compared with the water group in (B). DSS, dextran sulfate sodium; HBSS, Hanks’ balanced salt solution; PCC, peritoneal cavity cell; Veh, vehicle; Suc, succinate.