title: The efficacy and mechanism of apoptosis induction by hypericin-mediated sonodynamic therapy in THP-1 macrophages.
abstract: OBJECTIVE
  To investigate the sonoactivity of hypericin (HY), together with its sonodynamic effect on THP-1 macrophages and the underlying mechanism.
  
  
  METHODS
  CCK-8 was used to examine cell viability. Confocal laser scanning microscopy was performed to assess the localization of HY in cells, reactive oxygen species (ROS) generation, and opening of the mitochondrial permeability transition pore (mPTP) after different treatments. Apoptosis was analyzed using Hoechst-propidium iodide and transmission electron microscopy. Mitochondrial membrane potential (ΔΨm) collapse was detected via fluorescence microscopy. Lipoprotein oxidation was determined in malondialdehyde (MDA) assays. Western blotting was conducted to determine the translocation of BAX and cytochrome C and the expression of apoptosis-related proteins.
  
  
  RESULTS
  HY was sublocalized among the nuclei and the mitochondria, endoplasmic reticulum, Golgi apparatus, and lysosome in the cytosol of THP-1 macrophages. Under low-intensity ultrasound irradiation, HY significantly decreased cell viability and induced apoptosis. Furthermore, greater ROS generation, higher MDA levels, and greater ΔΨm loss were observed in the sonodynamic therapy (SDT) group. Both ROS generation and MDA levels were significantly reduced by the ROS scavenger N-acetyl cysteine (NAC) and the singlet oxygen scavenger sodium azide. Most of the loss of ΔΨm was inhibited by pretreatment with NAC, sodium azide, and the mPTP inhibitor cyclosporin A (CsA). mPTP opening was induced upon SDT but was reduced by pretreatment with bongkrekic acid, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid disodium, CsA, and NAC. Western blot analyses revealed translocation of BAX and cytochrome C, downregulated expression of Bcl-2, and upregulated expression of cleaved caspase-9, cleaved caspase-3, and cleaved poly(ADP-ribose) polymerase in the SDT group, which were reversed by NAC.
  
  
  CONCLUSIONS
  HY mediated SDT-induced apoptosis in THP-1 macrophages via ROS generation. Then, the proapoptotic factor BAX translocated from the cytosol to the mitochondria, increasing the ratio of BAX/Bcl-2, and the mPTP opened to release cytochrome C. This study demonstrated the great potential of HY-mediated SDT for treating atherosclerosis.
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Xuesong
authors: X
authors: Li
authors: Electron Microscopy Centre, Harbin Medical University, Harbin, People's Republic of China.
authors: Lei
authors: L
authors: Gao
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Longbin
authors: L
authors: Zheng
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Jiayuan
authors: J
authors: Kou
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Xing
authors: X
authors: Zhu
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Yueqing
authors: Y
authors: Jiang
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Zhaoyu
authors: Z
authors: Zhong
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Juhua
authors: J
authors: Dan
authors: Division of Cardiology, The First Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China.
authors: Haobo
authors: H
authors: Xu
authors: Division of Cardiology, The First Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China.
authors: Yang
authors: Y
authors: Yang
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Hong
authors: H
authors: Li
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Sa
authors: S
authors: Shi
authors: Laboratory of Sono- and Photo-theranostic Technologies, Harbin Institute of Technology, Harbin, People's Republic of China ; Materials Research Institute, The Pennsylvania State University, University Park, PA, USA.
authors: Wenwu
authors: W
authors: Cao
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Yajun
authors: Y
authors: Zhao
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China ; Division of Cardiology, The First Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China.
authors: Ye
authors: Y
authors: Tian
authors: Department of Pathophysiology, Harbin Medical University, Harbin, People's Republic of China.
authors: Liming
authors: L
authors: Yang
mesh_terms: D000873:Anthracenes; D017209:Apoptosis / Q000187:drug effects* / Q000528:radiation effects; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000473:pathology* / Q000528:radiation effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology*; D010569:Perylene / Q000031:analogs & derivatives* / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D014464:Ultrasonic Therapy
chemical_list: D000873:Anthracenes; D011838:Radiation-Sensitizing Agents; D017382:Reactive Oxygen Species; D010569:Perylene; C004965:hypericin; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D053453:Caspase 9
keywords: apoptosis; atherosclerosis; hypericin; mitochondria–caspase pathway; sonodynamic therapy
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