FIGURE 5.

Effect of gut microbiota and their products on cardiomyocyte apoptosis and colonic macrophage phenotypes in the doxorubicin (DOX)-treated mice. A single dose of DOX (20 mg/kg) was intraperitoneally injected into the C57BL/6 mice to induce acute cardiotoxicity. GLA (30 mg/kg) was intragastrically administered once daily for 12 days, starting 7 days before DOX injection. The mice from GLA (30 mg/kg) group were injected intraperitoneally with LPS (1 mg/kg), and orally administrated with sodium butyrate (1 g/kg) or Desulfovibrio vulgaris (Des) (1× 10⁹ CFU/mouse/every 2 days) 1 h after the injection of DOX. (A) The relative levels of D. vulgaris (n = 10). (B) Schematic representation of the experimental approach. (C) The relative levels of D. vulgaris (n = 10). (D) The levels of Lipopolysaccharide (LPS) and butyrate levels in feces and peripheral blood were determined by Limulus Amebocyte Lysate (LAL) assays or high performance liquid chromatography (HPLC), respectively (n = 10). (E) Relative mRNA levels of M1 marker (iNOS and CXCL-9) and M2 markers (CD206 and arginase-1) in the colonic macrophage were measured by qPCR (n = 10). (F) Immunostaining analysis of M1-like (CD68⁺CD206⁺) or M1-like (CD68⁺iNOS⁺) macrophages in the colon was shown (n = 10). Scale bars, 20 μm. (G) Corresponding quantitative analysis of the ratio of M2-like/M1-like macrophages (n = 10). (H) Serum levels of CK-MB AST, LDH, and CK were assessed (n = 10). (I) Representative images of the TUNEL assay and quantitative result (n = 10). Scale bars, 20 μm. The values are presented as the mean ± SEM. ^#P < 0.05, ^##P < 0.01 vs. control, ^∗P < 0.05, ^∗∗P < 0.01 vs. DOX, ˆP < 0.05, ˆˆP < 0.01 vs. DOX + GLA.