Xihuang Pill Induces Apoptosis of Human Glioblastoma U-87 MG Cells via Targeting ROS-Mediated Akt/mTOR/FOXO1 Pathway

Figure 5

ROS generation and mitochondrial dysfunction were required for XHP-induced U-87 MG cells apoptosis. (a) 2 × 10⁵ U-87 MG cells were treated with PTX or XHP for 24h and incubated with 10 mM JC-1 in darkness at 37°C for 30min, then ΔΨm was measured with FACS. Detection wavelengths were 530 nm for the green fluorescence and 585 nm for the red fluorescence. (b) Quantified data in JC-1 absorbed cells (n = 3, mean ± SD). (c) Cells were pretreated with or without ROS inhibitors (5 mM NAC) for 30 min, then were exposed to 30 µg/mL XHP for 0.5h, 1h or 3h, and then were collected and stained with DCFH-DA for measurement of H₂O₂ by FACS. H₂O₂ (100 mM) was used as positive control. (d) Cells were pretreated with ROS inhibitors (5 mM NAC) for 30min and then treated with the indicated procedure. FACS was employed to evaluate apoptosis. Each experiment was performed in triplicate.

(a)

(b)

(c)

(d)