ROS-Dependent Activation of Autophagy through the PI3K/Akt/mTOR Pathway Is Induced by Hydroxysafflor Yellow A-Sonodynamic Therapy in THP-1 Macrophages

Figure 2

HSYA-SDT induced autophagy in THP-1 macrophages. (a) Morphological alterations of THP-1 macrophages were observed using TEM. Red arrows indicate the autophagosomes (scale bar: 2 μm). (b) Autophagic vacuoles induced by different treatments were observed using MDC staining of HSYA-SDT-treated cells with or without 3-MA (10 mM) pretreatment. THP-1 macrophages were incubated with MDC (50 μM) for 30 min at 30 min after HSYA-SDT (scale bar: 20 μm). (c) LC3-Ι, LC3-ΙΙ, p62, and beclin 1 protein expression was analyzed by Western blots at different time points after HSYA-SDT, and quantifications of the LC3-ΙΙ/LC3-Ι ratio, p62, and beclin 1 are shown. (d) LC3-Ι, LC3-ΙΙ, p62, and beclin 1 protein expression was analyzed by Western blots at different time points after HSYA-SDT, and quantifications of the LC3-ΙΙ/LC3-Ι ratio, p62, and beclin 1 are shown. (e) Cells were stained with an anti-LC3 antibody and DAPI at 30 min after HSYA-SDT (scale bar: 5 μm). (f) The effect of 3-MA, hydroxychloroquine, and ba A1 with or without HSYA-SDT on the expression levels of the autophagy-related proteins p62 and LC3 and quantifications of the LC3-ΙΙ/LC3-Ι ratio and p62 are shown. All data are mean ± standard error (). , , and versus Control.

(a)

(b)

(c)

(d)

(e)

(f)