FIGURE 3.

Morphine-ADVEs induce the upregulation of miR-23a in pericytes via ADEV-mediated transporting miR-23a from astrocytes to pericytes. (A) ADEVs isolated from astrocytes treated with or without morphine for indicated time points, followed by assessing miR-23a expression using real-time PCR. (B,C) Pericytes and astrocytes were treated with or without morphine for 24 h, followed by an assessment of pre- (B) or mature-miR-23a (C) expression using real-time PCR. (D,E) miR-23a was up-regulated in the brains of morphine-dependent macaques as assessed by (D) real-time PCR and, (E) in situ hybridization. (F) Exposure of astrocytes to morphine for 24 h, followed by coculturing with pericyte for an additional 24 h. Then extracted RNA from pericytes, followed by an assessment of pre- or mature-miR-23a in the pericytes. (G) mouse primary astrocytes were transfected with mCherry TSG101 plasmid for 24 h, followed by coculturing with pericytes for an additional 24 h. Paraformaldehyde fixed pericytes were permeabilized and stained for pericyte marker PDGFR-β (Green) and visualized by fluorescence microscopy. Scale bars, 20 μm. All data are presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group using one-way ANOVA analysis.