高压脉冲电场联合低温等离子体对小鼠肝癌细胞杀伤效果的研究  

Combination effects of pulsed electric field and low temperature plasma on H22 mouse liver cancer cell

作　　者：胡笑钏 张晓伟 刘样溪 周古翔 吕毅[2] Hu Xiaochuan;Zhang Xiaowei;Liu Yangxi;Zhou Guxiang;Lyu Yi(Department of Electronic Control and Engineering,Chang’an University,Xi’an 710064,China;National and local Joint Engineering Research Center for Precision Surgery&Regenerative Medicine,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]长安大学电子与控制工程学院,西安710064 [2]西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心,西安710061

出　　处：《中华肝胆外科杂志》2022年第5期373-378,共6页Chinese Journal of Hepatobiliary Surgery

基　　金：中国博士后科学基金(2021M702629);国家自然科学基金(81727802)。

摘　　要：目的探究高压脉冲电场(PEF)联合低温等离子体(LTP)对小鼠肝癌细胞杀伤效果。方法小鼠肝癌细胞H22分为肝癌细胞组(无任何处理)、PEF处理组、LTP处理组、联合A组(先行PEF处理后立即行LTP处理)、联合B组(先行LTP处理后立即行PEF处理)、联合C组(同联合A组,但间隔20 min)、联合D组(同联合B组,但间隔20 min)。细胞计数试剂盒检测细胞存活率,流式细胞仪检测凋亡,荧光标记细胞内活性氧(ROS)并计数。20只4~6周龄健康雌性无特定病原体的昆明小鼠建立皮下移植瘤模型,随机分为模型组、PBS对照组、PEF实验组、LTP实验组及联合组(LTP+PEF,无间隔),每组4只。测量并计算肿瘤相对体积和抑瘤率。结果肝癌细胞组细胞存活率(98.3±0.9)%,PEF处理组(66.8±4.4)%,LTP处理组(62.1±3.9)%,联合A组(43.7±3.7)%,联合B组(31.0±1.4)%,联合C组(46.8±2.9)%,联合D组(39.0±2.3)%。与肝癌细胞组比较,各处理组细胞存活率均降低,且四个联合处理组细胞存活率低于PEF处理组和LTP处理组,差异均有统计学意义(均P<0.05)。其中联合B组细胞存活率最低。凋亡检测结果与细胞存活率结果一致。荧光显微镜下,联合B组细胞ROS荧光明显增多,且LTP处理组细胞ROS荧光比PEF处理组多。联合B组的ROS阳性细胞百分比高于LTP处理组和PEF处理组,差异均有统计学意义(均P<0.05)。联合组抑瘤率和肿瘤相对体积优于PEF实验组和LTP实验组,差异均有统计学意义(均P<0.05)。结论LTP联合PEF对小鼠肝癌细胞杀伤效果优于PEF和LTP单一处理,有望成为新的肿瘤治疗方法。Objective To investigate the effect of pulsed electric field(PEF)combined with low temperature plasma(LTP)on mouse liver cancer cell.Methods H22 mouse liver cancer cells were divided into liver cancer group,PEF treatment group,LTP treatment group,combined group A(first PEF treatment immediately after LTP treatment),combined group B(first LTP treatment immediately after PEF treatment),combined group C(same as combined group A,but 20 minutes interval)and combined group D(same as combined group B,but 20 minutes interval).Cell viability was detected by cell counting,apoptosis was detected by flow cytometry,intracellular reative oxygen species(ROS)was marked by fluorescence and counted.Twenty healthy female Kunming mouse aged 4-6 weeks without specific pathogens were subcutaneous injected with liver cancer cells,and then were randomly divided into model group,PBS control group,PEF experimental group,LTP experimental group and combined group(LTP+PEF,no interval)(n=4).Tumor relative volume and tumor inhibition rate were measured.Results The survival rates were liver cancer cell group(98.3±0.9)%,PEF treatment group(66.8±4.4)%,LTP treatment group(62.1±3.9)%,combined group A(43.7±3.7)%,combined group B(31.0±1.4)%,combined group C(46.8±2.9)%,combined group D(39.0±2.3)%.Compared with liver cancer cell group,the cell survival rate of all treatment groups was decreased,and the cell survival rate of the four combined treatment group was lower than that of PEF treatment group and LTP treatment group,the differences were statistically significant(all P<0.05).The survival rate of combined B group was the lowest.The results of apoptosis detection were consistent with those of cell survival rate.Under fluorescence microscope,the ROS fluorescence of cells in the combined group B was significantly increased,and the ROS fluorescence of cells in the LTP treatment group was more than that in the PEF treatment group,and the percentage of ROS positive cells in the combined group B was higher than that in the LTP treatment group and

关 键 词：活性氧 高压脉冲电场 低温等离子体 肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]