Fig. 5
Suppression of CX3CR1high fibrocytes by pravastatin (PRA) inhibits colitis-associated carcinogenesis. (a) Treatment scheme of the azoxymethane/dextran sodium sulfate (AOM/DSS) model with or without PRA. (b) The number of neoplasms at 20 weeks after AOM injection was compared between control and PRA-treated CX3CR1 wild-type (WT)-bone marrow (BM) and CX3CR1 knockout (KO)-BM chimeric mice (n = 12/group). ***P < 0.001 by a one-way ANOVA. (c) Flow cytometry representative density and dot plots of the colonic lamina propria (LP) of AOM/DSS-injured CX3CR1 WT-BM chimeric mice treated with or without PRA. Lineage (Ly6G, Siglec F, B220, CD3)−CD11b+ macrophages from the LP of CX3CR1 WT-BM chimeric mice treated with or without PRA were divided into GFP-CX3CR1high macrophages (P1), GFP-CX3CR1low monocytes (P2 & P3), macrophages (P4), and dendritic cells (P5).
(d) The absolute cell numbers for the five fractions in the colonic LP of control and PRA-treated mice are presented (n = 6/group). **P < 0.01 by an unpaired Student’s t-test.
(e) The proportion of GFP-CX3CR1+CD45+CD11b+ColI+ fibrocytes among CD45+ cells in the colonic LP of control and PRA-treated mice is presented (n = 5/group). **P < 0.01 by an unpaired Student’s t-test. (f) In vitro apoptosis assay of BM monocytes isolated from CX3CR1 WT and CX3CR1 KO mice. The left panel depicts the experimental scheme. Monocytes, which were incubated in medium containing the control or PRA, were treated with lipopolysaccharide for 24 h. The middle panel presents the proportion of apoptotic cells (7-AAD+ cells) compared between control and PRA-treated CX3CR1 WT and CX3CR1 KO mice (n = 6/group). The right two panels show the expression of mRNA of Bcl2l1 and Bax compared between control and PRA-treated CX3CR1 WT and CX3CR1 KO mice. *P < 0.05, ***P < 0.001, and ****P < 0.0001 by a one-way ANOVA. Data are presented as the mean ± SD.
