Fig 2: Effect of curcumin on M1 and M2 macrophage marker protein expression in the myocardium at 7 days post-MI. (a) Representative immunohistochemistry detection of M1 macrophage marker (iNOS) expression; (b) representative immunohistochemistry detection of M2 macrophage marker (Arg1) expression; (c) quantification of iNOS-positive cells; (d) quantification of Arg1-positive cells. Seven mice in each group. Data were shown as the mean ± SEM; ∗ was P < 0.05 vs. the sham group, and # was P < 0.05 vs. the MI group. And P value was calculated by Student's t-test.

Fig 3: NEU1 deletion promotes the polarization of macrophages. (A,B) Gating strategy for CD45+CD11b+F4/80+ CD86+ M1 macrophages and CD45+CD11b+ F4/80+ CD206+ M2 macrophages in aortas of NEU1 CKO mice and their controls. (C,D) Quantification of the CD86+ and CD206+ macrophages. (E,F) Immunofluorescence images of iNOS+ and Arg-1+ macrophages in aortas of NEU1F/F and NEU1 CKO mice subjected to BAPN induction for 4 weeks (scale bar: 50 μm). (G,H) mRNA levels of M1 and M2 markers in macrophages sorted by flow cytometry of NEU1F/F and NEU1 CKO mice subjected to BAPN induction for 4 weeks (n = 6, *p < 0.05). (I,J) Macrophages sorted from NEU1F/F and NEU1 CKO mice, and subjected to Ang II treatment for 24 h. Immunofluorescence images of iNOS+ and Arg-1+ macrophages by Ang II treatment (scale bar: 50 μm). Ang, angiotensin; Arg-1, arginase-1; BAPN, β-aminopropionitrile monofumarate; DHE, dihydroethidium; iNOS, inducible nitric oxide synthase; KO, knockout; NEU1, neuraminidase 1; NEU1 CKO, macrophage-specific NEU1 knockout.

Fig 4: MiR-520a-3p silencing inhibited macrophage polarization to M1 subset. (A) RT-PCR analysis showed the mRNA levels of macrophage polarization related markers TNF-α; (B) IL-6; (C) MCP-1; (D) IL-1β; (E) INOS; (F) CD206; and (G) Arg1 in plaque and artery tissue in apoE-/- mice. Data are mean ± SEM; one-way ANOVA was used for the statistical analysis. n=5 mice per group; (H) Western blot analysis showed increased CD206 and decreased INOS after miR-520a-3p inhibition in plaque and artery tissue in apoE-/- mice. Data are mean ± SEM; one-way ANOVA was used for the statistical analysis. n=5 mice per group. **P<0.01, ***P<0.001.

Fig 5: ROSI declined COM-stimulated M1Mp polarization and crystal adhesion and promoted M2Mp polarization in vitro. (a) Fluorescence immunohistochemical distribution of Arg1 (green) and iNOS (red) in THP-1 cells with the stimulus of COM or ROSI (1 μM) or GW9662 (10 μM) (arrows). (b) Evaluation of mean fluorescence intensity (MFI) of Arg1 and iNOS. (c) ELISA. The expression levels of IL-4, IL-10, IL-6, and TNF-α. (d) Crystal adhesion assay. COM crystal adhesion to HK-2 cells was observed (arrows). COM: calcium oxalate monohydrate; ROSI: rosiglitazone; iNOS: induced nitric oxide synthase; TNF-α: tumor necrosis factor α. ∗p < 0.05; ∗∗p < 0.01.