Fig. 3

Microglia polarized into the proinflammatory M1 type, accompanied by an increase in TNF‐α in the RVLM of SIH rats. A Representative images of Iba1 immunofluorescence and the Z-stack projection of microglia in the RVLM of control and SIH rats. Scale bar = 20 or 5 μm. B Skeletal microglia analysis of total process length, the number of branching points, and the number of terminal points in control and SIH rats. C, D Sholl analysis of RVLM microglial processes in control and SIH rats. E Representative immunoblot bands and quantitative analysis showed the expression of iNOS, CD86, and Arg-1 in the RVLM between the two groups. F Immunofluorescence co-localization analysis of TNF-α in the RVLM. Iba1, GFAP, and NeuN were used as markers for microglia, astrocytes, and neurons, respectively. Scale bar = 50 μm. G, H The TNF-α expression was determined by RT-qPCR and Western blot assays in the RVLM of control and SIH rats. Data were expressed as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t-test (A, B, E, G, H) and two-way repeated measures ANOVA (C, D). n = 30 microglia from 6 rats (A–D). n = 3 rats per group (E, H). n = 6 rats per group (G). *p < 0.05,  **p < 0.01, ***p < 0.001 vs. Control group