Fig. 2 | Signal Transduction and Targeted Therapy

Fig. 2

From: Porphyromonas gingivalis aggravates atherosclerotic plaque instability by promoting lipid-laden macrophage necroptosis

Fig. 2The alternative text for this image may have been generated using AI.

Pg infection induces lipid-laden macrophage necroptosis enlarging necrotic core. a Non-parametric Spearman’s test between the ratio of necrotic area and the amount of Pg positive clusters within the human coronary atherosclerotic plaques. r = 0.7902, P < 0.0001. n = 33. b–g Quantification of the proportion of dying macrophages probed by TUNEL and CD45-CD68/F4/80 co-staining in human coronary plaques (b, c) (group low, n = 7; group medium, n = 7, group high, n = 6), rabbit aortic arch plaques (d, e) (n = 6 per group) and mouse aortic root plaques (f, g) (n = 6 per group). Nuclei were labeled using DAPI. Scale bar = 100 μm on top, and 20 μm on the bottom. h H&E, Masson, and Oil Red O staining of atherosclerotic plaques in aortic roots of Apoe−/− mice untreated or infected with Pg and/or administrated with vehicle or liposomal clodronate (LC) for 8 weeks. Scale bar = 200 μm in H&E, scale bar = 100 μm in the rest images. i Quantitative analyses of plaque size, necrotic area, collagen content, Oil Red O+ area, thickness of fibrous cap of the plaques. n = 6 per group. j The ratio of PI+ cells in macrophages over a time course of stimulation with or without ox-LDL (60 μg/mL) and/or Pg (MOI = 100). n = 4 per group. k Flow cytometry analyses of the ratio of necrotic (PI+Annexin V+) and apoptotic (PI−Annexin V+) macrophages under ox-LDL (60 μg/mL) and/or Pg (MOI = 100) treatment. n = 4 per group. l The ratio of PI+ cells in ox-LDL loaded macrophages pre-administrated with Z-VAD-FMK (5 μM), Nec-1 (10 μM), ferrostatin-1 (FER-1, 5 μM), or VX765 (50 μM) and infected with or without Pg (MOI = 100). n = 4 per group. The co-localization of p-MLKL and CD68 or F4/80-labled macrophages in the atherosclerotic plaques of human coronary arteries (m), rabbit aortic arches (n), and mouse aortic roots (o). Nuclear DNA (blue) was counterstained with DAPI. Scale bar = 100 μm. Results were represented as mean ± SEM (c, e, g, i) or mean ± SD (j, k, l). Data were analyzed by non-parametric Spearman’s test (a), one-way ANOVA (c, i, k), unpaired Student t test (e) and two-way ANOVA (g, j, l). ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05

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