超声辐照对HepG_2细胞存活率影响的实验研究

Effect of ultrasound irradiation on viability of HepG_2 cell:an experimental study in vitro

为探讨超声辐照对体外培养的HepG2细胞存活力的影响,采用正交设计方法,选定声强、占空比和辐照时间3因素,每因素5水平的参数条件设计正交表,对贴壁培养的HepG2细胞进行超声辐照,之后继续培养24 h后行MTT(噻唑蓝)实验检测细胞存活率,期间光学显微镜观察培养板内的细胞生长状况及形态。结果表明:当超声声强大于2 W/cm2任何情况及声强1.5 W/cm2、占空比大于60%时,细胞出现明显脱壁、形态变圆、弹坑样聚集以及脱壁细胞内出现颗粒和细胞碎裂等现象,细胞存活率大大降低。超声声强在0.5～1.0 W/cm2,占空比在10%-20%时,细胞存活率可确保达到90%以上。正交设计统计分析提示超声声强和占空比是影响细胞存活率的主要因素,且声强影响大于占空比(P<0.05),而辐照时间并非主要因素(P>0.05)。

To investigate the effect of ultrasound irradiation on viability of HepG2 cells in vitro,the parameters of acoustic intensity,duty factor and duration of therapeutic ultrasound are selected,and the ultrasound irradiations are performed according to the orthogonal design.After 24 h of ultrasound irradiations,MTT assays are performed to assess the viability of HepG2 cells.The morphologic change of cells are observed using optical micrography before and after ultrasound irradiation.The results show that,when the acoustic intensity is greater than 2 W/cm2 with any duty factor or the acoustic intensity is 1.5 W/cm2 with the duty factor greater than 60%,the viability of HepG2 cells decreases significantly,and the detachment,craterlike aggregation,and cell fragmentation of cells could be observed by optical microscope.When the acoustic intensity ranges from 0.5 to 1.0 W/cm2 and the duty factor is 10%-20%,the rate of cell viability can be more than 90% immovably.The statistical analysis of orthogonal design indicates that the acoustic intensity and duty factor are the most important parameters for the HepG2 cell viability irradiated by therapeutic ultrasound in vitro.