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. 2021 Aug 19;11:16806. doi: 10.1038/s41598-021-94708-1

Figure 1.

Figure 1

Aβ oligomers exposition induces AD pathological signatures and microglia activation which are totally rescued by AZP2006 treatment. (A, B) After 72 h of Aβ1-42 exposure, the number of rat cortical neurons, culture with microglia, is reduced (MAP-2 staining; CTR = 49 positive soma, ± 2, n = 6) and dendritic morphology changed to a less complex network (CTRL = 15,365 µm/well ± 200, n = 6). The presence of AZP2006 in the same medium rescue neurons survival and morphology. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by Dunnett’s test, n = 4–6. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition. (C) Neurons were stained with postsynaptic marker PSD95 and presynaptic marker synaptophysin; their overlap was automatically quantified (overlapping PSD95/SYN, μm2 of overlapping) to obtain the total synapses area. CTR = 242 µm2/well, ± 1.5, n = 5. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by PLSD Fisher’s test, n = 4–5. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition. (D) The immunostaining quantification for OX-41, an antibody clone which specifically recognizes macrophages and granulocytes, show microglia activation, reduced after AZP2006 treatment. CTR = 3134 µm2/well, ± 197.3 n = 5. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by PLSD Fisher’s test, n = 5–6. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition. (E, F) Released IL-1β and IL-6 protein quantification (supernatant of cell culture) from rat cortical neurons and microglia exposed to Aβ1-42 oligomers for 72 h with or without AZP2006. CTR IL-1β = 0.065 pg/ml/cell, ± 0.005, n = 5 ; CTR IL-6 = 0.10 4 pg/ml/cell, ± 0.008, n = 4. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by PLSD Fisher’s test, n = 4–6. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition. (G) AT100 area in neurites of cortical neurons cultured with microglia, injured with Aß1-42 (5 µM, 72 h) and treated with AZP2006. CTR = 1.751 µm2 (mean per neurite/well), ± 0.13, n = 5. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by PLSD Fisher’s test, n = 4–6. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition. (H) After 72 h of treatment, neuron lysates were removed and analyzed for PGRN protein level. CTR = 1.917 ng/ml, ± 0.109, n = 6. All values are expressed as mean ± SEM (standard error of the mean). One-way ANOVA followed by PLSD Fisher’s test, n = 5–6. #p < 0.05 versus CTR. *p < 0.05 versus Aβ1-42 condition.