高强度聚焦超声治疗对裸鼠皮下胶质瘤VEGF、PCNA表达以及细胞凋亡的影响  

The effect of high intensity focused ultrasound on VEGF and PCNA expression and apoptosis of subcutaneous neurogliocytoma in nude mice

作　　者：肖文峰[1] 李俊[1] 霍钢[2] 翟安林[1] 郑履平[2] 

机构地区：[1]川北医学院附属第二医院神经外科,绵阳621000 [2]重庆医科大学附属第一医院神经外科

出　　处：《中国神经精神疾病杂志》2015年第7期416-421,共6页Chinese Journal of Nervous and Mental Diseases

基　　金：重庆市卫生局资助项目(编号:04-2-196)

摘　　要：目的通过观察高强度聚焦超声(high intensity focused ultrasound,HIFU)对裸鼠皮下人胶质瘤模型作用后血管内皮生长因子(vascular endothelial growth factor,VEGF)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)蛋白的表达和细胞的凋亡,探讨HIFU治疗后对交界区及照射区细胞增殖和细胞凋亡的影响。方法建立18只裸鼠左右背部皮下胶质瘤模型,用频率9.7MHz,焦距4.5mm,声强2500W/cm2的HIFU连续照射20s。按照射后动物处死时间随机分为7d组,14d组,30d组,每组各6只裸鼠即12个皮下胶质瘤组织;应用免疫组化法检测HIFU作用后第7d、14d、30d照射区、交界区、正常区VEGF、PCNA蛋白的表达;应用原位末端标记法(Td T-mediated d UTP nick end labeling,TUNEL)检测照射区、交界区、正常区的凋亡细胞。结果免疫组化观察到,各时刻点交界区及正常区均有VEGF及PCNA表达并检测到凋亡染色阳性的细胞,照射区均无PCNA蛋白的表达及未检测到凋亡细胞。第7d时交界区VEGF阳性细胞百分数(23.79%±3.11%)低于正常区(46.16%±2.43%)(F=110.03,P<0.05);照射区VEGF阳性细胞百分数(10.94%±3.95%)低于正常区(46.16%±2.43%)(F=272.80,P<0.05)。第14d时交界区VEGF阳性细胞百分数(17.17%±2.89%)低于正常区(43.47%±3.77%)(F=152.05,P<0.05);第30d时交界区VEGF阳性细胞百分数(9.27%±2.08%)低于正常区(44.58%±3.34%)(F=274.1,P<0.05 2)。交界区的PCNA标记指数(PCNA labeling index,PCNA LI)(33.04%±4.31%)在第7d时低于正常区(65.15%±3.85%),(F=242.46,P<0.05);在14d时为(21.05%±1.96%)低于正常区(62.99%±3.34%)(F=413.52,P<0.05);在30d时为(6.36%±0.51%)低于正常区(62.07%±18.07%)(F=729.59,P<0.05)。第7d时交界区的凋亡指数(apoptotic index,AI)(26.10%±4.54%)高于正常区(1.43%±0.35%)(F=216.22,P<0.05)。14d时交界区的凋亡指数AI(65.70%±1.14%)高于正常区(1.82%±0.31%)(F=1448.64,P<0.05);30d时交界区的凋亡指数AI(82.02%±3.98%)高于正常区(2.52%±0.29%)(F=2244.33,P<0.05)。�Objective To explore the effects of high intensity focused ultrasound （HIFU） on cell multiplication and apoptosis at exposure coverage and marginal zone and the expression of vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen and apoptosis of subcutaneous neurogliocytoma in nude mice. Methods Eighteen nude mice bearing subcutaneous human neurogliocytoma were consecutively ablated in 20s by an extracorporeal HIFU with 9.7MHz transducer （the focal length of 4.5mm and focal intensity 2500W/cm^2）. The 18 nude mice were randomly divided into 7 d group, 14 d group and 30 d group according to sacrifice date. Immunohistochemical method, TdT-mediated dUTP nick end labeling method were used to examine the expression of vascular endothelial growth factor and proliferating cell nuclear antigen and apoptosis at exposure coverage, marginal zone and normal zone, respectively. Results The expression of VEGF and proliferating cell nuclear antigen were evident at exposure coverage, marginal zone and normal zone in 7, 14 and 30 days after ablation. The expression of proliferating cell nuclear antigen and apoptosis were absent at exposure coverage in 7,14 and 30 days after ablation. The percentage of VEGF expression was lower in marginal zone than in normal zone （23.79%±3.11% vs. 46.16%±2.43%） in 7 d after ablation （F=110.03,P〈0.05）. The percentage of VEGF expression was also lower （10.94%±3.95%） in exposure coverage than in normal zone （46.16%±2.43%） in 7 d after ablation （F=272.80,P〈0.05）. The percentage of VEGF expression was lower in marginal zone than in normal zone （17.17%±2.89% vs. 43.47%±3.77%） in 14 d after ablation （F=152.05,P〈0.05）. The percentage of VEGF expression was lower in marginal zone than in normal zone （9.27%±2.08% vs. 44.58%±3.34%） in 30 d after ablation （F=274.1,P〈0.05 2）. The proliferating cell nuclear antigen labeling index（PCNA LI） was lower in marginal zone than in normal zone （（33.04%±4.31% vs. 65.15%±3

关 键 词：高强度聚焦超声 胶质瘤 细胞增殖 细胞凋亡 

分 类 号：R739.41[医药卫生—肿瘤]