Fig. 4
Astrogliosis in SDH of remote segments mediated widespread pain. A, B Immunofluorescence staining showed increased IBA1 expression in CDH, TDH, and LDH of CCI-ION rats. Scale bar: 50 μm. t test, *p < 0.05, n = 6/group. C, D Immunofluorescence staining showed increased GFAP expression in CDH, TDH, and LDH of CCI-ION rats. Scale bar: 50 μm. t test, *p < 0.05, **p < 0.01, n = 6/group. E The workflow of intraspinal application LAA, minocycline or vehicle. F The time course of mechanical escape threshold of vibrissal pad for CCI-ION rats receiving LAA, minocycline or vehicle. Two-way ANOVA, n = 8/group. G, H The paw thermal withdrawal latency and mechanical withdrawal threshold in CCI-ION rats receiving LAA, minocycline or vehicle. Two-way ANOVA, **p < 0.01, ***p < 0.001, ****p < 0.0001, n = 8/group. I Heatmap showed Ca2+ response of primary cultured astrocytes to CSF from CCI-ION rats and sham rats. J Quantitative analysis showed the percentage of responsive astrocytes after addition of CSF from CCI-ION rats and sham rats. Chi-square test, ****p < 0.0001. K, L Immunofluorescence staining of C3 in primary cultured astrocytes after incubation of sham’s CSF and CCI-ION’s CSF. t test, ****p < 0.0001, n = 9/group. Scale bar: 75 μm. M, N Immunofluorescence staining of Ki67 in primary cultured astrocytes after incubation of sham’s CSF and CCI-ION’s CSF. t test, ****p < 0.0001, n = 9/group. Scale bar: 75 μm. O, P Immunofluorescence staining showed GFAP expression in CDH, TDH, and LDH of rats receiving sham’s CSF or CCI-ION’s CSF. t test, ****p < 0.0001. n = 10–14/group. Scale bar: 50 μm