title: Real-time detection of intracellular reactive oxygen species and mitochondrial membrane potential in THP-1 macrophages during ultrasonic irradiation for optimal sonodynamic therapy.
abstract: Reactive oxygen species (ROS) elevation and mitochondrial membrane potential (MMP) loss have been proven recently to be involved in sonodynamic therapy (SDT)-induced macrophage apoptosis and necrosis. This study aims to develop an experimental system to monitor intracellular ROS and MMP in real-time during ultrasonic irradiation in order to achieve optimal effect in SDT. Cultured THP-1 derived macrophages were incubated with 5-aminolevulinic acid (ALA), and then sonicated at different intensities. Intracellular ROS elevation and MMP loss were detected in real-time by fluorospectrophotometer using fluorescence probe DCFH-DA and jc-1, respectively. Ultrasound at low intensities (less than 0.48W/cm(2)) had no influence on ROS and MMP in macrophages, whereas at an intensity of 0.48W/cm(2), ROS elevation and MMP loss were observed during ultrasonic irradiation. These effects were strongly enhanced in the presence of ALA. Quantitative analysis showed that ROS elevation and MMP loss monotonically increased with the rise of ultrasonic intensity between 0.48 and 1.16W/cm(2). SDT at 0.48 and 0.84W/cm(2) induced mainly apoptosis in THP-1 macrophages while SDT at 1.16W/cm(2) mainly cell necrosis. This study supports the validity and potential utility of real-time ROS and MMP detection as a dosimetric tool for the determination of optimal SDT.
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China; Department of Pathophysiology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, Harbin 150081, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Sun
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China.
authors.forename: Haobo
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Oncology, The Third Affiliated Hospital, Harbin Medical University, Harbin 150081, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Shen
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China.
authors.forename: Shuyuan
authors.initials: S
authors.lastname: Guo
authors.affiliation: Department of Pathophysiology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, Harbin 150081, China.
authors.forename: Sa
authors.initials: S
authors.lastname: Shi
authors.affiliation: Department of Pathophysiology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, Harbin 150081, China.
authors.forename: Juhua
authors.initials: J
authors.lastname: Dan
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Cardiovascular Institute, Harbin Medical University, Harbin 150001, China; Department of Pathophysiology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, Harbin 150081, China. Electronic address: yetian@ems.hrbmu.edu.cn.
authors.forename: Ye
authors.initials: Y
authors.lastname: Tian
mesh_terms: D000622:Aminolevulinic Acid / Q000378:metabolism; D017209:Apoptosis / Q000528:radiation effects; D002460:Cell Line; D045744:Cell Line, Tumor; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism* / Q000528:radiation effects; D008264:Macrophages / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects*; D053078:Membrane Potential, Mitochondrial / Q000528:radiation effects*; D009336:Necrosis; D010789:Phototherapy; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014464:Ultrasonic Therapy
chemical_list: D017382:Reactive Oxygen Species; D000622:Aminolevulinic Acid
keywords: 5-Aminolevulinic acid; Macrophage; Mitochondrial membrane potential; Reactive oxygen species; Sonodynamic therapy