Blue Laser Irradiation Generates Intracellular Reactive Oxygen Species in Various Types of Cells
Blue Laser Irradiation Generates Intracellular Reactive Oxygen Species in Various Types of Cells
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DOI：
10.1089/pho.2012.3361
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发表时间：
2013-03-01
影响因子：
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通讯作者：
Ishihara, Miya
中科院分区：
文献类型：
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作者：
Kushibiki, Toshihiro;Hirasawa, Takeshi;Ishihara, Miya
目标：本研究的目的是测量激光照射不同类型细胞后细胞内活性氧（ROS）的产生。背景资料：ROS被认为是低强度激光治疗（LLLT）产生的关键第二信使。虽然LLLT的作用机制多种多样，但细胞内活性氧（ROS）是影响LLLT作用的关键因素之一，但直接测定不同波长激光照射后几种细胞内ROS的研究尚未见报道。材料与方法：本研究中使用了各种类型的细胞：小鼠前脂肪细胞（3 T3-L1）、前软骨细胞（ATDC 5）、成肌细胞（C2 C12）、间充质基质细胞（KUSA-A1）、肺癌细胞（LLC）、胰岛素瘤细胞（MIN 6）、成纤维细胞（NIH-3 T3）、人宫颈腺癌细胞（HeLa）、用佛波醇酯处理后从淋巴细胞分化的巨噬细胞（THP-1），和大鼠嗜碱性白血病细胞（RBL 2 H3）。用蓝色激光（波长：405 nm）、红色激光（波长：664 nm）或近红外激光（波长：808 nm）以100 mW/cm（2）照射细胞60秒或120秒。在流式细胞仪中使用细胞内ROS探针CM-H（2）DCFDA通过荧光测定法测量细胞内ROS水平。结果：蓝光照射后，细胞内ROS水平在所有类型的细胞中均增加。相反，用红色激光或近红外激光照射后未观察到细胞内ROS的产生。结论：蓝色激光照射激发了细胞内潜在的ROS源，导致细胞内ROS的产生。虽然低水平的细胞内活性氧应产生后，红色或近红外激光照射，只有高水平的细胞内活性氧检测到的ROS探针在本研究中使用。由于活性氧被认为是关键的第二信使，激光照射对细胞的特定功能调节将在未来的研究中进行研究。
Objective: The purpose of this study was to measure intracellular reactive oxygen species (ROS) production after laser irradiation in various types of cells. Background data: ROS are considered to be the key secondary messengers produced by low-level laser therapy (LLLT). Although various mechanisms for the effects of LLLT have been proposed, and intracellular ROS were indicated as the one of the key factors, direct measurement of intracellular ROS of several types of cells after different wavelength lasers irradiation has not been reported. Materials and methods: Various types of cells were used in this study: mouse preadipocytes (3T3-L1), pre-chondrocytes (ATDC5), myoblasts (C2C12), mesenchymal stromal cells (KUSA-A1), lung cancer cells (LLC), insulinoma cells (MIN6), fibroblasts (NIH-3T3), human cervix adenocarcinoma cells (HeLa), macrophages differentiated from lymphocytes (THP-1) after treatment with phorbol ester, and rat basophilic leukemia cells (RBL2H3). Cells were irradiated with a blue laser (wavelength: 405 nm), a red laser (wavelength: 664 nm) or a near infrared laser (wavelength: 808 nm) at 100 mW/cm(2) for 60 or 120 sec. Intracellular ROS levels were measured by fluorometric assay using the intracellular ROS probe, CM-H(2)DCFDA in a flow cytometer. Results: After a blue laser irradiation, intracellular ROS levels were increased in all types of cells. In contrast, intracellular ROS generation was not observed after irradiation with a red laser or near-infrared laser. Conclusions: Potential sources of intracellular ROS were excited by blue laser irradiation, resulting in ROS production within cells. Although the low-level intracellular ROS should be generated after a red or a near-infrared laser irradiation, the only high level intracellular ROS were detected by the ROS probe used in this study. As ROS are considered to be key secondary messengers, the specific functional regulation of cells by laser irradiation will be studied in a future study.