Journal: Journal of Neuroinflammation
Article Title: Targeting glial fibrillary acidic protein in glaucoma: a monoclonal antibody approach to modulate glial reactivity and neuroinflammation for neuroprotection
doi: 10.1186/s12974-025-03482-8
Figure Lengend Snippet: Microarray analysis reveals the effects of GFAP mAb treatment on key inflammatory mediators in glaucomatous retina. ( A - C ) The expression of TLR4 and S100A8 was significantly downregulated by 25–50 µg GFAP mAb treatment compared to the vehicle. The marker for microglial activation, CD68, was significantly decreased in the 25 µg GFAP mAb treatment group. ( D - F ) Proteins of the inflammasome pathway, NLRP3, GSDMD and Caspase-1 were significantly downregulated in the 25 µg GFAP mAb treatment group. With the 50 µg dose, these protein expressions showed a similar downward trend, but without statistical significance. ( G ) Expression profiling of inflammation-associated mediators showed that 25 µg GFAP mAb significantly decreased pro-inflammatory factors (TNF-α, IL-1β, IL-8, MMP9, and IFN-γ), and increased the anti-inflammatory cytokine IL-10 compared to the vehicle group. In contrast, the 50 µg dose significantly reduced IFN-γ only. ( H ) Representative images of spots showing IL-1β levels in the subarrays for each group. Image analysis was conducted with Imagene software, using the median intensity from each spot to calculate the mean of the triplicate spots for each marker. For CD68 and TLR4, due to significant heterogeneity in SDs, statistical analysis was performed using Welch’s ANOVA followed by the Tamhane T2 post hoc test. All other data were analyzed using one-way ANOVA with Tukey’s post hoc test. Data are presented as the mean ± SD; n = 4 per group; * p < 0.05, ** p < 0.01, *** p < 0.001; ns = not significant. TLR4 – Toll-Like Receptor 4. S100A8 – S100 Calcium Binding Protein A8. NLRP3 – NLR Family Pyrin Domain Containing 3. GSDMD – Gasdermin D
Article Snippet: Briefly, the microarray was prepared using a noncontact microarray printer (SciFLEXARRAYER S3; Scienion, Berlin, Germany).
Techniques: Microarray, Expressing, Marker, Activation Assay, Software, Binding Assay
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