Fig. 6

Ex vivo treatment of microglial cells from 5XFAD mice with Malva parviflora hydroalcoholic extract increases the microglia phagocytic activity. a Microglia was purified from adult (wild type and 5XFAD) mouse brains as described in the “Methods” section. Microglia enrichment was determined by flow citometry using anti-CD11b antibodies. A representative histogram depicts isotype (red) and CD11b (blue) labeled cells from 5XFAD transgenic brain. b Primary microglial cells were purified from 8- and 10-months-old Wt and 5XFAD mice as described in the “Methods” section and left untreated (Ctrl) or treated with the M. parviflora extract (1 mg/mL) for 24 h and then exposed to fluorescent E. coli (red) for 4 h. Cells were fixed and stained with phalloidin and DAPI for F-actin (green) and nuclei (blue) detection, respectively. Microglia was imaged using the Olympus FluoView 1000 confocal multiphoton microscope (Scale bar, 30 μm). Insets show a × 2 digital magnification from boxed areas. c Graph depicts the percentage of cells with internalized E. coli (m.o.) that were left untreated (−) or treated (+) with the M. parviflora extract (1 mg/mL) for 24 h and then exposed to fluorescent E. coli (red) for 4 h. Data (mean ± SEM) were analyzed by one-way ANOVA followed by Tukey’s post hoc test (^*p < 0.05). d Average of microorganisms (E. coli) internalized by microglia from the indicated mouse strain and age untreated (−) or treated (+) with the M. parviflora extract (intervals were from 1 to 5, from 6 to 10 and greater than 10 microorganisms/cell). Data (mean ± SEM) were analyzed by paired t test (^**p < 0.01; ^***p < 0.001). e Phagocytic index was calculated by multiplying the percentage of microglia with internalized bacteria by the average number of internalized E. coli bacteria per each group. Data were collected from seven random fields per group and analyzed by one-way ANOVA followed by Tukey’s post hoc test (^***p < 0.001)