[1]谢霜,杨敏,杜永洪,等.低频低强度超声对巨噬细胞内耻垢分枝杆菌的损伤效应[J].第三军医大学学报,2018,40(16):1476-1482.
 XIE Shuang,YANG Min,DU Yonghong,et al.Low-frequency and low-intensity ultrasound suppresses activity of Mycobacterium smegmatis in macrophages[J].J Third Mil Med Univ,2018,40(16):1476-1482.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第16期
页码:
1476-1482
栏目:
基础医学
出版日期:
2018-08-30

文章信息/Info

Title:
Low-frequency and low-intensity ultrasound suppresses activity of Mycobacterium smegmatis in macrophages
作者:
谢霜杨敏杜永洪李岱容
重庆医科大学:生物医学工程学院,省部共建国家重点实验室培育基地—重庆市超声医学工程重点实验室,重庆市生物医学工程学重点实验室,重庆市微无创医学协同创新中心,附属第一医院呼吸科
Author(s):
XIE Shuang YANG Min DU Yonghong LI Dairong

School of Biomedical Engineering, State Key Laboratory of Ultrasound Engineering in Medicine CoFounded by Chongqing and the Ministry of Science and Technology, College of Biomedical Engineering, Chongqing Key Laboratory of Biomedical Engineering, Chongqing Collaborative Innovation Center for Minimallyinvasive and Noninvasive Medicine, Chongqing Medical University;  Department of Respiratory Diseases, First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
低频低强度超声巨噬细胞耻垢分枝杆菌凋亡活性氧
Keywords:
low-frequency and lowintensity ultrasound macrophages Mycobacterium smegmatis apoptosis reactive oxygen species
分类号:
R378.91; R392.12; R454.3
文献标志码:
A
摘要:

目的   探讨低频低强度超声(low frequency low intensity ultrasound,LFLIU)对巨噬细胞内耻垢分枝杆菌的损伤效应及机制。方法    将巨噬细胞RAW264.7分为超声辐照0、5、10 min和15 min组(n=3),应用MTT比色法分析不同超声强度对RAW264.7细胞存活率的影响,采用流式细胞术检测RAW264.7细胞经超声辐照以及吞噬细菌后的凋亡率和坏死率,使用平板计数法评估RAW264.7细胞内耻垢分枝杆菌活性,使用激光共聚焦显微镜观察RAW264.7细胞经超声辐照后活性氧(reactive oxygen species,ROS)的产生。结果    超声辐照声强为0.14 W/cm2,辐照时间为5、10 min时,RAW264.7细胞活性未见明显影响,存活率分别为(116.37±9.54)%、(97.14±9.26)%;经0.14 W/cm2超声辐照5 min后,RAW264.7细胞的凋亡率和坏死率显著大于超声辐照0 min组(P<0.05);平板菌落计数结果显示,RAW264.7细胞内耻垢分枝杆菌经声强0.14 W/cm2超声辐照5、10 min后活性均低于超声辐照0 min组(P<0.05);激光共聚焦观察到 0.14 W/cm2超声刺激RAW264.7细胞产生了ROS。结论    LFLIU 能诱导RAW264.7细胞凋亡和产生ROS,并导致RAW264.7细胞内耻垢分枝杆菌活性下降。

Abstract:

Objective    To investigate the effect of low-frequency and low-intensity ultrasound (LFLIU) in inducing damages of Mycobacterium smegmatis (M. smegmatis) in macrophages and explore the mechanism. Methods    Murine macrophage RAW264.7 cells were exposed to ultrasound irradiation for 5, 10 or 15 min, and the changes in the cell viability were analyzed with MTT assay. The apoptotic and necrosis rates of RAW264.7 cells after ultrasonic irradiation and after phagocytosis of M. smegmatis were assessed using flow cytometry; the activity of M. smegmatis in RAW264.7 cells following the exposure was evaluated with plate colony counting. Laser scanning confocal microscopy was used to observe the production of reactive oxygen species (ROS) in RAW264.7 cells after ultrasound exposure. Results    Exposure of RAW264.7 cells to a ultrasonic intensity of 0.14 W/cm2 for 5 min and 10 min resulted in cell survival rates of (116.37±9.54)% and (97.14±9.26)%, respectively, suggesting no significant impact of LFLIU exposure on the cell viability. The apoptotic rate of RAW264.7 cells increased significantly after irradiation with 0.14 W/cm2 ultrasound for 5 min compared with the control cells (P<0.05). The results of plate colony counting suggested that the viability of M. smegmatis in RAW264.7 cells was significantly lowered after exposure to a ultrasound intensity of 0.14 W/cm2 for 5 and 10 min. Confocal laser scanning microscopy revealed obvious ROS production in RAW264.7 cells in response to 0.14 W/cm2 ultrasound exposure. Conclusion    Exposure to LFLIU induces apoptosis and ROS production in RAW264.7 cells and causes decreased activity of M. smegmatis within the cells.

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更新日期/Last Update: 2018-09-03