Table 1.
Distribution and proportion of mNSS scores of mice post BE.
Fig 1.
Electrophysiological and behavioral assessments of visual dysfunction following blast wave exposure.
(A) Schematic illustration of the preparation procedure and observation time points for the bTBI animal model. (B) Schematic diagram of F-VEP recording device (C) Representative F-VEP waveforms at BE 24h, BE 3d, BE 7d, and BE 28d, the horizontal grid is 50 ms and the vertical grid is 10 μV. Quantitative analysis of P2 latency (D) and N2-P2 amplitude (E) (n = 30). (F) Continuous F-VEP recording (n = 12), the shaded area indicates mean ± SD. (G) Schematic of the bright chamber and dark chamber in the Light-dark box test. (H) Heatmap (left figure) and trajectory (right figure) visualization of mouse positions in different groups. (I) Quantitative analysis of stay time in dark areas in different groups (n = 14). Error bars indicate mean ± SD. ** p < 0.01, *** p < 0.001, ns p > 0.05.
Fig 2.
Temporal Progression of Blast-Induced Optic Nerve Demyelination.
(A) Schematic diagram of the visual pathway and key observation areas in its extracranial segment. (B) Representative images of immunofluorescence staining for MBP (green) and βIII-Tubulin (red) in mouse optic nerve slices. Group information is shown in images, scale bar = 20 μm. (C) Optic nerve relative fluorescence intensity of MBP +/β III Tubulin + between the sham group and BE 24h, BE 3d, BE 7d, BE 28d groups, n = 4. (D-E) Western blot analysis of MBP in optic nerve lysates. The molecular mass is indicated in kilodaltons, n = 3. (F) TEM analysis of the optic nerve between the sham group and BE 24h and BE 28d. Red stars indicate axons with obvious demyelination. Error bars indicate mean ± SD. ***p < 0.001.
Fig 3.
Temporal Progression of Blast-Induced Pyroptosis in Visual Cortex Neurons.
(A) Schematic diagram of the visual pathway and key observation areas in its intracranial segment. (B) Representative images of immunofluorescence staining for NeuN (green) and NLRP3 (red), Iba-1 (grey) in mouse visual cortex slices, Group information is shown in images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20μm. Quantitative analysis of visual cortex immunofluorescence density in Iba-1 (C) and NLRP3 (G). (H) Fluorescence intensity of NeuN+NLRP3+/ NeuN+, n = 4. (D, E, F, I, J) Western blot analysis of Iba-1, IL-1β, cleaved Gasdermin D and NLRP3 in visual cortex lysates, the molecular mass is indicated in kilodaltons, n = 3. (K-L) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (M) Quantitative analysis of visual cortex NeuN+ cells, n = 4. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.
Fig 4.
MCC950 Attenuates Blast-Induced Visual Dysfunction.
(A) Representative F-VEP waveforms between the BE 24h, BE 24h + DMSO, and BE 24h + MCC950 groups, the horizontal grid is 50 ms and the vertical grid is 10 μV. (B-C) Quantitative analysis of P2 latency and N2-P2 amplitude (n = 20). (D) Representative F-VEP waveforms between the BE 28d, BE 28d + DMSO, and BE 28d + MCC950 groups. (E-F) Quantitative analysis of P2 latency and N2-P2 amplitude (n = 20). (G) Representative heatmap (left figure) and trajectory (right figure) visualization of mouse positions in different groups. (H-I) Quantitative analysis of stay time in dark areas in different groups (n = 12). All group information is shown in images. Error bars indicate mean ± SD. ***p < 0.001, ns p > 0.05.
Fig 5.
MCC950 Attenuates Neuroinflammation and Significantly Suppresses Pyroptosis, But Doesn’t Attenuate Optic Nerve Demyelination.
(A) Representative images of immunofluorescence staining for NeuN (green), NLRP3 (red), and Iba-1 (grey) in mouse visual cortex slices, as well as group information, are shown in the images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20 μm, n = 4. (B) Quantitative analysis of visual cortex immunofluorescence density in NLRP3, n = 4. (C) Fluorescence intensity of NeuN+NLRP3+/NeuN+, n = 4. (D) Quantitative analysis of visual cortex immunofluorescence density in Iba-1, n = 4. (E, F, G, H) Western blot analysis of Iba-1, cleaved Gasdermin D and NLRP3 in visual cortex lysates. The molecular mass is indicated in kilodaltons, n = 3. (I, J) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (K) Quantitative analysis of visual cortex NeuN+ cells. (L) Representative images of immunofluorescence staining for MBP (green) and βIII-Tubulin (red) in mouse optic nerve slices. Group information is shown in images. Scale bar = 20 μm, n = 4. (M) optic nerve MBP density in different groups, n = 4. (N) optic nerve relative fluorescence intensity of MBP +/β III Tubulin + in different groups, n = 4. (O) TEM analysis of the optic nerve between the BE 24 h and BE 28 d groups, red stars indicate axons with obvious demyelination. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.