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Fig. 11 | Journal of Neuroinflammation

Fig. 11

From: Trazodone regulates neurotrophic/growth factors, mitogen-activated protein kinases and lactate release in human primary astrocytes

Fig. 11

TDZ modulation of the AKT pathway. a, b Astrocytes were treated with medium alone (basal), or with the indicated concentrations of TDZ, or FLUOX (10 μM) or 5-HT (10 μM) for 30 min (a) or 24 h (b). Following incubation, the levels of phosphorylated and total AKT were evaluated using an ELISA kit as described in the Methods section. The data are expressed as phosphorylated/total AKT ratio. The data are the mean ± SEM of three independent experiments performed in triplicate. c Cells were pre-incubated with medium alone (basal), or 15 nM (S)-WAY 100135 (5-HT1AR antagonist), or 30 nM (R)-DOI (5-HT2A/CR agonist), or 250 nM clonidine (α-AR agonist), or 100 μM histamine (H1 histamine receptor agonist). After 15 min, cells were incubated with TDZ (10 μM) for an additional 30 min. d Human astrocytes were pre-treated with medium alone (basal), or 15 nM (S)-WAY 100135 (5-HT1AR antagonist), or 10 nM GR 127935 (5-HT1B/DR antagonist), or 5 nM RS 127445 (5-HT2BR antagonist), or 30 nM (R)-DOI (5-HT2A/CR agonist) or 100 nM SR 57227 (5-HT3R agonist). After 15 min, cells were incubated with FLUOX (10 μM) for an additional 30 min. Following incubation, the levels of phosphorylated and total AKT were evaluated using an ELISA kit. e Human astrocytes were pre-treated with 200 ng/ml PTX (Gαi/o inhibitor), or 1 μM H89 (PKA inhibitor), or 1 μM bisindolylmaleimide (PKC inhibitor); then, cells were incubated with TDZ (10 μM) or FLUOX (10 μM) for an additional 30 min. Following incubation, the levels of phosphorylated and total AKT were evaluated using an ELISA kit. The significance of the differences was determined using a one-way ANOVA-Tukey HSD post hoc test: *P < 0.05, **P < 0.01, ***P < 0.001 vs. basal; # P < 0.05, ## P < 0.01 vs cells stimulated with TDZ or FLUOX

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