Fig 2: In vivo immunomodulatory effectiveness of BD@Zn-MSN-MN. (A) Illustrative H&E-stained spleen sections across varied groups. (B) Representative histological analysis of skin tissue sections using toluidine blue. (C) Examples of immunofluorescent staining for CD206 and iNOS in skin tissues on day 7. (D) Exemplary immunofluorescent staining depicting CGRP in skin tissue sections on day 7. (E) Mast cell count analysis (n = 6 per group, ns: not significant, *p < 0.05, ****p < 0.0001, analyzed using one-way ANOVA with Tukey’s post-hoc test). (F) Ratio of MFI between CD206 and iNOS in mice skin (n = 6 per group, **p < 0.01, ****p < 0.0001, analyzed using one-way ANOVA with Tukey’s post-hoc test). (G) MFI assessment of CGRP in mice skin (n = 4 per group, ns: not significant, ****p < 0.0001, analyzed using one-way ANOVA with Tukey’s post-hoc test). (H-J) Relative mRNA expression levels of Il-17f (H), Il-23 (I), and Tnf-α (J) in mouse skin (n = 6 per group, ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, analyzed using one-way ANOVA with Tukey’s post-hoc test)

Fig 3: Effect of HIV-1 Tat treatment on NF-kappaB activation in the nuclear extracts of frontal cortex and astrocytes activation. (a) The panel shows representative NF-kappaB activation complex bands in the different groups of rats. (b) The quantitative analysis revealed that intracolonic HIV-1 Tat administration yields to a significant increase of NF-kappaB, as compared to vehicle, lidocaine, or bisacodyl groups (OD = optical density in mm2). (c) HIV-1 Tat caused a marked increase of GFAP, S100B, TLR-4 and iNOS protein expression in the frontal cortex homogenates of treated rats. (d) Quantitative analysis reveled that HIV-1 Tat induced a significantly higher expression of GFAP, S100B, TLR-4 and iNOS, than lidocaine, or bisacodyl groups. (e,f) In the medium of frontal cortex homogenates deriving from HIV-1 Tat group a significant increase of NO2 − and S100B was also observed as compared to the other groups. (Results are expressed as mean ± SEM; ***p < 0.001 vs all other groups; °°°p < 0.001 vs HIV-1 Tat group; n = 6 for each group).

Fig 4: Intracolonic administration of HIV-1 Tat induced glial activation in the (a) thoracic and (b) cervical spinal cord and (c) frontal cortex at day 12, 14 and 21 after diarrhea induction, respectively. (a–c) Immunofluorescence analysis showed that iNOS (green) and S100B (red) co-expression was increased in the spinal cord and frontal of HIV-1 Tat treated rats. (d–f) Quantitative analysis showed that HIV-1 Tat-induced upregulation of iNOS (filled bars) and S100B (open bars) was significantly inhibited by lidocaine treatment. Results are expressed as mean ± SEM; ***p < 0.001 vs all other groups; °°°p < 0.001 vs HIV-1 Tat group. Scale bars: 100 μm; n = 6 for each group.

Fig 5: MSR1 knockout inhibited pro-inflammatory polarization of macrophages after treatment with myelin debris in vitro. a IF staining of F4/80 (green), iNOS (pro-inflammatory macrophage marker, purple), and CD206 (anti-inflammatory macrophage marker, red) in the lesion sites of the MSR1 WT and KO mice. Bar = 200 μm. b Determination of the pro-inflammatory and anti-inflammatory macrophages by analysis the number of iNOS+ and F4/80+ cells or CD206+ and F4/80+ cells in the lesion sites of the MSR1 WT and KO mice (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, two-tailed Student’s t tests). c Quantitative PCR of pro-inflammatory macrophage marker genes (iNOS, TNF-α, and IL-β) and anti-inflammatory macrophage marker genes (CD206, YM1/2, and Arg1) in the lesion sites of the MSR1 WT and KO mice at day 7 post injury (n = 3 per group, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA). d IF staining of iNOS and CD206 in MSR1 WT and KO macrophages after treatment with myelin debris for 24 h. Scale bar = 10 μm. e Quantitative PCR of pro-inflammatory macrophage marker genes (iNOS, TNF-α, and IL-1β) and anti-inflammatory macrophage marker genes (CD206, YM1/2, and Arg1) in MSR1 WT and KO macrophages before or after treatment with myelin debris (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA). f–g Flow cytometric analysis of myelin debris-treated macrophages from MSR1 WT or KO mice. The percentages of F4/80+ iNOS+ and F4/80+ CD206+ macrophages in the MSR1 WT and KO groups were calculated (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA)