Figure 6
Role of σ-1R in the activation of astrocytes mediated by methamphetamine. (A) Exposure of primary rat astrocytes to different concentrations (15 μM, 150 μM, and 1.5 mM) of methamphetamine increased the GFAP expression with maximal response at the concentration of 150 μM. (B) Effect of methamphetamine (150 μM) increased the expression of GFAP in rat primary astrocytes. (C) Pretreatment of primary rat astrocytes with σ-1R antagonist-BD1047 significantly inhibited methamphetamine-mediated increase in GFAP expression. (D) Immunofluorescent staining for GFAP (green) in primary rat astrocytes treated with methamphetamine and σ-1R antagonist-BD1047 (10 μM). Scale bars - 50 μm. (E) Quantification of GFAP immunofluorescent intensity from five areas on the slide using the Image J software. IOD-integrated optical density. (F) Pretreatment of primary rat astrocytes with Src inhibitor-PP2 (10 μM), MEK1/2 (U0126, 10 μM), or PKA inhibitor-H89 (10 μM), resulted in inhibition of methamphetamine-mediated expression of GFAP. Representative immunoblots and the densitometric analysis of σ-1R/GAPDH from the three separate experiments are presented. (G) Compared with primary mouse astrocytes from WT mice, methamphetamine failed to increase the expression of GFAP in astrocyte isolated from σ-1R KO mice. All the data are mean ± SD of three individual experiments. *P < 0.05 versus control group; #P < 0.05 versus methamphetamine-treated group in astrocytes from WT mice.