摘要
目的研究低强度超声结合5-氨基酮戊酸(5-ALA)对大鼠骨肉瘤UMR-106细胞的杀伤机制。方法将处于对数生长期的大鼠骨肉瘤UMR-106细胞分成对照组、5-ALA组、超声组和超声+5-氨基酮戊酸组(SDT组)。流式细胞仪检测细胞凋亡率,活性氧(ROS)的产生,线粒体膜电位(MMP)的改变;荧光显微镜观察33342染色细胞核的形态改变;透射电镜观察超微结构改变。结果当超声频率为1.0 MHz,声强2.0 W/cm2,5-ALA浓度2 mmol/L,SDT组与对照组、超声组及5-ALA组比较,其凋亡率(32.2±1.4)%,明显增高(P<0.05)。同时伴有ROS(34.4±2.4)%的产生和MMP(42.2±2.6)%的降低。通过33342染色荧光显微镜观察到超声联合5-ALA组的细胞核发生浓缩和碎裂。透射电镜观察到细胞膜、线粒体、高尔基体等细胞器改变及凋亡小体的形成。结论低强度超声联合5-ALA对UMR-106细胞的杀伤作用明显,细胞以凋亡为主,其线粒体途径对UMR-106细胞凋亡起着重要作用。
Objective To investigate the killing effect of low -intensity ultrasound combined with 5-aminolevulinic acid (5-ALA) on rat osteosarcoma cell line UMR -106.Methods After the UMR-106 cells came into logarithm growth period , they were divided into control group and experimental group .Cell apoptotic rate ,the production of reactive oxygen species (ROS) and the change of mitochondrial membrane potential ( MMP) was observed by flow cytometry;ultrastructural changes were observed by transmission electron microscope ( TEM) .Results Using low intensity ultrasound of 1.0 MHz and 2.0 W/cm2 , plus 5-ALA at a concentration of 2 mmol/L.The apoptotic rate of SDT group was (32.2 ±1.4)%which was signifi-cantly higher than that of control group, ultrasound group and 5 -ALA group(P &lt;0.05).The production of ROS was (34.4 ±2.4)%and the decrease of MMP was (42.2 ±2.6)%.Through the 33342 staining was observed in ultrasound com-bined with 5-ALA group of nuclear condensation and fragmentation .The structural changes of cell membrane, mitochondria Golgi apparatus and other organelles could be observed by TEM clearly Formation of apoptotic bodies .Conclu sion The killing effect of low-intensity ultrasound combined with 5-ALA on UMR-106 cells was significantly .Cell apoptosis played a vital role in the killing effect, and the mitochondria pathway contributed mainly in the apoptosis of UMR -106 cells.
出处
《大连医科大学学报》
CAS
2014年第3期216-220,共5页
Journal of Dalian Medical University
基金
中央高校基本科研业务费专项资金资助项目(HIT.NSRIF.2014066)