Figure 1. Effects of low-dose chronic treatment with I2-IR LSL60101, Donepezil and co-administration on cognitive status in 5XFAD and WT mice. (a) Scheme of experimental design. Results of NORT: Discrimination index calculated by using exploration time for novel and familiar object (b) in the short-term term memory test session (2h) (WT, n=10-12 per group; 5XFAD n=11-12 per group)(c) in the long-term memory test session (24h) (WT, n=10-12 per group; 5XFAD n=10-12 per group). Results of MWM : (d)Distance to target (platform) (cm) during the training session (WT n=10-11 per group; 5XFAD n=10-12 per group) (e) Quadrant preference in the test session as time (%) spent in each quadrant WT n=9-12 per group; 5XFAD n=10-12 per group) (f) Time (%) spent in platform quadrant in the test session (WT n=9-12 per group; 5XFAD n= 10-12 per group) (g) Distance to target (platform) (cm) in the test session (WT n=9-12 per group; 5XFAD n=10-12 per group) (h)Representative images of trajectory during memory test. Bars represent mean±SEM. one-way and two-way ANOVA with Tukey post hoc analysis, *P<0.05.
Figure 2. Effects of low-dose chronic treatments I2-IR LSL60101, Donepezil and co-administration on behavioral and social status in 5XFAD mice and WT controls. Results of OF: (a) Locomotor activity measured as distance travelled (cm) WT n=10-12 per group; 5XFAD n=11-12 per group(b) Time spent in the center (sec) (WT n=10-12 per group; 5XFAD n=9-11 per group) (c) number of rearings (WT n=11-12 per group; 5XFAD n=10-12 per group). Results of EPM: (d) Time (%) spent in Open Arms (WT n=11-12 per group; 5XFAD n=10-12 per group)(e) Time (%) spent in closed arms (WT n=11-12 per group; 5XFAD n=10-12 per group) (f) number of rearings (WT n= 11-12 per group; 5XFAD n=11-12 per group). Results of TCT: (g) Entries in chambers (A1, A2) in the habituation phase (n) (WT n=11-12 per group; 5XFAD n=11-12 per group) (h) Time in chambers (Empty, Stranger) during the test session (WT n=10-11 per group; 5XFAD n=9-11 per group)(i) time of interaction with intruder (sec) in the test session (WT n=11-12 per group; 5XFAD n=10-11 per group). Bars represent mean±SEM. Student’s t-test or two-way ANOVA with Tukey post hoc analysis, *P<0.05;
Figure 3. Effects of low dose chronic treatments I2-IR LSL60101, donepezil and co-administration on AD hallmarks in 5XFAD mice. (a)Histological images of amyloid plaques stained with Thioflavin-S and quantification of (b) number of the plaques in cortex and hippocampus (c ) and area (%) covered by plaques in the DG area of the hippocampus in the 5XFAD mice. Representative Western blot and quantifications (d-f) for β-Amyloid p-Tau(Ser404) and p-Tau(ser396) in the hippocampus of 5XFAD mice. Values in bar graphs are adjusted to 100% for protein levels of the control WT control or the control 5XFAD. Bars represent mean±SEM. Student’s t-test or one-way ANOVA with Tukey post hoc analysis, *P<0.05; n=5-6 per group.
Figure 4. Effects of low dose chronic treatments I2-IR LSL60101, donepezil and co-administration on Aβ pathology and APP processing in 5XFAD mice. Representative Western Blots and Quantifications for(a-g) CTFs, p-APP, sAPPβ, sAPPα, BACE 1, ADAM10 in the hippocampus of 5XFAD mice. Values in bar graphs are adjusted to 100% for protein levels of the control WT control or the control 5XFAD. Representative gene expression in the hippocampus of the 5XFAD mice for(h) APP and (i) IDE, Nep. Gene expression levels were determined by real-time PCR. Bars represent mean±SEM. Student’s t-test or one-way ANOVA with Tukey post hoc analysis, *P<0.05; n=5-6 per group.
Figure 5. Effects of low dose chronic treatments I2-IR LSL60101, donepezil and co-administration on microgliosis and inflammatory markers in 5XFAD mice. (a) Representative images of GFAP immunostaining and quantification (b-d) in DG, CA1, CA3 areas of the hippocampus of the 5XFAd mice. Representative gene expression in the hippocampus of the 5XFAD mice for (e) Il-1β, Il-6, Ccl12, (f)for Trem2 . Gene expression levels were determined by real-time PCR. Bars represent mean±SEM. Student’s t-test or one-way ANOVA with Tukey post hoc analysis, *P<0.05; n=5-6 per group.
Figure 6. Effects of low dose chronic treatments I2-IR LSL60101, donepezil and co-administration on astrogliosis and synaptic plasticity in 5XFAD mice. (a) Representative images of GFAP immunostaining and quantification (b-d) in DG, CA1, CA3 areas of the hippocampus of the 5XFAd mice. Representative western blot and quantifications(f-h) for PSD95 and SYN in the hippocampus of 5XFAD mice. Values in bar graphs are adjusted to 100% for protein levels of the control WT control or the control 5XFAD. Bars represent mean±SEM. Student’s t-test or one-way ANOVA with Tukey post hoc analysis, *P<0.05; n=5-6 per group.