The mTOR/GCLc/GSH Pathway Mediates the Dose-Dependent Bidirectional Regulation of ROS Induced by TiO₂ NPs in Neurogenic Cells

Figure 2

(a) mTOR and p-mTOR (Ser2448) protein expressions were measured by western blotting experiment after cells were treated with 1 μg/ml and 100 μg/ml of TiO₂ NPs for 24 h. (b) ROS levels after cells were treated with 1 μg/ml TiO₂ NPs and 1 μg/ml TiO₂ NPs+RAPA. Mean fluorescence intensity was measured by FCM. (c) GCLc protein expression levels after cells were treated with 1 μg/ml and 100 μg/ml of TiO₂ NPs for 24 h. (d) GCLc protein expressions after cells were treated with 1 μg/ml TiO₂ NPs and 1 μg/ml TiO₂ NPs+RAPA. (e) GSH, GSSG levels, and the ratio of GSH/GSSG were measured by a commercial assay kit after cells were treated with 1 μg/ml and 100 μg/ml TiO₂ NPs for 24 h, and the results were from three panel samples of each experiment. Each assay was repeated at least three times. (f) The H₂O₂ concentrations after cells were treated with TiO₂ NPs for 24 h. Total protein was used as an internal control. (g, h) Intracellular superoxide anion was measured by DHE after cells were treated with TiO₂ NPs for 24 h. The independent experiment was repeated three times and five times, respectively. GAPDH was used as protein loading control. Blank means blank control; “0” means control group; RAPA means rapamycin. All the values are presented as , , , .

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