Journal: Signal Transduction and Targeted Therapy
Article Title: Porphyromonas gingivalis aggravates atherosclerotic plaque instability by promoting lipid-laden macrophage necroptosis
doi: 10.1038/s41392-025-02251-6
Figure Lengend Snippet: Oxidative stress serves as a principal driver of Pg –induced necroptosis. a Dihydroethidium (DHE) staining and CD45-ACTA2 co-staining of serial sections of the aortic roots isolated from Apoe –/– mice infected with or without Pg for 8 weeks. Nuclei were labeled using DAPI. Scale bar = 100 μm. b The co-localization of NOX2 and CD45-labled macrophages in the aortic root plaques of Apoe –/– mice infected with or without Pg for 8 weeks. Nuclei were labeled using DAPI. Scale bar = 20 μm. c Flow cytometry analyses of the reactive oxygen species (ROS) level in macrophages infected by Pg in different MOIs, and treated with or without ox-LDL (60 μg/mL). n = 4 per group. d Relative mRNA expression levels of Nox2 , Cox2 , Nos2 , Gpx1 , Sod1 , and Sod2 in ox-LDL (60 μg/mL)-loaded macrophages treated with or without Pg (MOI = 100) for 24 h. n = 4 per group. Flow cytometry analyses of ROS production ( e ) and ratio of PI + cells ( f ) in ox-LDL (60 μg/mL)-loaded macrophages pre-administrated with EUK134 (10 μM) and infected with Pg (MOI = 100). n = 4 per group. g Western blot analyses of RIPK3, p-MLKL, MLKL, and GAPDH expression in ox-LDL (60 μg/mL)-loaded macrophages pre-administrated with EUK134 (10 μM) and infected with Pg (MOI = 100) for 24 h. GAPDH was used as the loading control. h H&E, Masson, Oil Red O staining, and CD45 and F4/80 co-immunohistochemical staining of atherosclerotic plaques in aortic roots of Apoe −/− mice untreated or challenged by Pg under the administration of EUK134 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, Oil Red O + area, and CD45 and F4/80-positive areas of the plaques in h . n = 6 per group. Results were represented as mean ± SD ( c , d , e , f ) or mean ± SEM ( i ). Data were analyzed by two-way ANOVA ( c , d ), or one-way ANOVA ( e , f , i ). **** P < 0.0001; *** P < 0.001; ** P < 0.01; * P < 0.05
Article Snippet: To investigate the effect of oxidative stress on Pg -promoted atherosclerotic plaque destabilization, EUK134 (10 mg/kg, S4261, Selleck) or DMSO (as control) was delivered via intraperitoneal injection to Apoe −/− mice once per week, combined with or without intravenous injection with 10 7 CFU Pg once per week for 8 weeks.
Techniques: Staining, Isolation, Infection, Labeling, Flow Cytometry, Expressing, Western Blot, Control, Immunohistochemical staining
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