Fig 2: Long-term MO treatment alleviates neuroinflammation in APP/PS1 mice. (A) The expression level of GFAP and IBA1 from hippocampal lysates of WT control and APP/PS1 mice with or without MO treatment was evaluated by Western blotting, (n = 3). (B,C) The statistical analysis of Western blot bands from panel A. ß-actin serves as the loading control. (D,E) The statistical analysis of TNF-a and IL-1ß ELISA results from hippocampal lysates of WT control and APP/PS1 mice with or without MO treatment was evaluated, (n = 4). The data are presented as Mean ± SEM. ** p < 0.01 and *** p < 0.001 vs. WT or vs. APP/PS1 mice groups.

Fig 3: The effect of tDCS on the main components of the NVU. (A,B) Representative images and statistical analysis of NeuN, Iba1, GFAP, LRP1 and PDGRFß expression in the hippocampus (A) and frontal cortex (B) in each group. n = 3. The data are expressed as the mean ± SEM. #, compared with the WT group. *, compared with the ADT group. *P < 0.05, **P < 0.005, ###/***P < 0.001.

Fig 4: Microglia trended to the pro-inflammation stage after SNI. (A) Western blot showed that the expression of TNF-a and CD86 was significantly increased in the spinal dorsal cord at 7 and 14 days after SNI. (B) The expression of Arg-1 was decreased in the spinal dorsal cord at 3, 7, and 14 days, and the expression of CD163 was significantly decreased at 3 and 7 days after SNI. (*p < 0.05, **p < 0.01, ***p < 0.001 compared with sham group, n = 6 in each group). (C) Double immunostaining of CD86 (green) and Iba1 (red) in the spinal dorsal horn. The arrows show the representative merge of CD86 and Iba1. (D) Double immunostaining of CD163 (green) and Iba1 (red) in the spinal dorsal horn. The merge of CD163 and Iba1 is represented by arrows. Scale bars, 100 µm for lower magnification and 50 µm for higher magnification. (E) Histograms are quantifications of the percentage of CD86+-Iba1+ cells and CD163+-Iba1+ cells. Results of each group were obtained from three rats, and three micrographs from each sample. The average values of each sample were taken for statistics. (*p < 0.05, **p < 0.01 compared with sham group). (F) The expression of TNF-a mRNA, CD86, and iNOS mRNA was significantly increased and Arg-1, CD163, and IL-10 mRNA was decreased in the spinal dorsal cords of SNI 7d rats (*p < 0.05, **p < 0.01, ***p < 0.001 compared with sham group, n = 4 in each group).

Fig 5: SNI induced hyperalgesia changed the expression and morphology of microglia in the spinal dorsal horn. (A,B) SNI significantly reduced the MWT (A) and improved the acetone score (B) in the ipsilateral paws. (****p < 0.0001 compared with the sham group, n = 12 in each group). (C) Double immunofluorescence of CB2R and Iba1 in the spinal cord of sham and SNI-7d rats. The fluorescence images in the boxes were magnified to better display the double staining. Scale bar, 100 µm. (D) The expression of Iba1 increased on days 3, 7, and 14 after SNI. (*p < 0.05 compared with the sham group, n = 6 in each group). (E) microglia micrographs from the ipsilateral spinal dorsal horn sham or SNI-7d rats. The images with higher magnification were skeletonized and expressed as black-and-white images (right side) for measuring the three parameters. Scale bars, 100 µm for lower magnification and 50 µm for higher magnification. (F) Quantification of the area of Iba1+ cells and morphology of microglia. Solidity, the area of cell body / the area of processes for each cell. Results for each group were obtained from three rats and three micrographs from each sample. The average values of each sample were taken for statistics. (**p < 0.01, ***p < 0.001 compared with the sham group).