Fig. 6

Chronic morphine disturbed host systemic immunity and induced microglial mitophagy in mice. A The quantitative analysis of organ indexes in mice (n =46). B The heat maps of liver (left), spleen (middle) and thymus (right) indicated the mean values of mRNA expression of inflammatory cytokine genes (n = 4–6). C Representative Western blots of Hsp60, Tim23 and LC3 in cortex, striatum, hippocampus, cerebellum and brainstem in control or morphine-treated mice (n = 3–4). D Confocal microscopy analysis of Iba1 (red), NLRX1 (green), LC3B (magenta) and DAPI (blue) in cortex, striatum and cerebellum in control or morphine-treated mice. Bar = 50 μm. E The ratios of NLRX1⁺ cells in microglia, NLRX1⁺ cells in LC3B + microglial cells and microglia in NLRX1 + cells in D were analyzed (n = 11–12 fields). Data represent the mean ± SEM. Two-sided Student’s t tests were used to measure significance between two groups (* p < 0.05, ** p < 0.01, *** p < 0.001 and ns p > 0.05)