Protoporphyrin IX-mediated sonodynamic therapy promotes autophagy in vascular smooth muscle cells

Sonodynamic therapy (SDT) is effective in treating intimal hyperplasia and promoting plaque stability in animal models. The present study aimed to evaluate the effects of SDT with the sonosensitizer protoporphyrin IX (PpIX) on vascular smooth muscle cell (VSMC) viability and autophagy. Cultured VSMCs cells were divided into the following groups: i) Control, ii) ultrasound, iii) PpIX and iv) SDT. Flow cytometry and laser confocal detection were used to measure Annexin V stained VSMCs following different treatments. Alterations in mitochondrial membrane potential (MMP) were evaluated via JC-1 staining. Autophagosome formation was observed using electron and fluorescence microscopy. Western blotting was used to analyze the expression levels of the autophagy markers light chain 3 (LC3-I) and LC3-II. The results demonstrated that SDT did not trigger apoptosis nor induce a significant decline in MMP of VSMCs. However, SDT significantly increased autophagasome formation and increased the LC3-II/LC3-I ratio. The findings demonstrated that PpIX-SDT increased autophagy without inducing mitochondrial-dependent apoptosis in VSMCs.