中国神经精神疾病杂志
中國神經精神疾病雜誌
중국신경정신질병잡지
CHINESE JOURNAL OF NERVOUS AND MENTAL DISEASES
2015年
7期
416-421
,共6页
肖文峰%李俊%霍钢%翟安林%郑履平
肖文峰%李俊%霍鋼%翟安林%鄭履平
초문봉%리준%곽강%적안림%정리평
高强度聚焦超声%胶质瘤%细胞增殖%细胞凋亡
高彊度聚焦超聲%膠質瘤%細胞增殖%細胞凋亡
고강도취초초성%효질류%세포증식%세포조망
Neurogliocytoma%High Intensity Focused Ultrasound%Cell Multiplication%Apoptosis
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 anti?gen，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(TdT-mediated dUTP 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<0.052)。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)。结论在本实验设定参数及声学剂量情况下, HIFU除了对照射区组织产生直接的杀灭作用外，还能抑制交界区的细胞增殖以及诱发细胞凋亡的发生。
目的：通過觀察高彊度聚焦超聲(high intensity focused ultrasound ，HIFU)對裸鼠皮下人膠質瘤模型作用後血管內皮生長因子(vascular endothelial growth factor，VEGF)、增殖細胞覈抗原(proliferating cell nuclear anti?gen，PCNA)蛋白的錶達和細胞的凋亡，探討HIFU治療後對交界區及照射區細胞增殖和細胞凋亡的影響。方法建立18隻裸鼠左右揹部皮下膠質瘤模型，用頻率9.7MHz，焦距4.5mm，聲彊2500W/cm2的HIFU連續照射20s。按照射後動物處死時間隨機分為7d組，14d組，30d組，每組各6隻裸鼠即12箇皮下膠質瘤組織；應用免疫組化法檢測HIFU作用後第7d、14d、30d照射區、交界區、正常區VEGF、PCNA蛋白的錶達；應用原位末耑標記法(TdT-mediated dUTP nick end labeling，TUNEL)檢測照射區、交界區、正常區的凋亡細胞。結果免疫組化觀察到，各時刻點交界區及正常區均有VEGF及PCNA錶達併檢測到凋亡染色暘性的細胞，照射區均無PCNA蛋白的錶達及未檢測到凋亡細胞。第7d時交界區VEGF暘性細胞百分數(23.79%±3.11%)低于正常區(46.16%±2.43%)(F=110.03,P<0.05)；照射區VEGF暘性細胞百分數(10.94%±3.95%)低于正常區(46.16%±2.43%)(F=272.80， P<0.05)。第14d時交界區VEGF暘性細胞百分數(17.17%±2.89%)低于正常區(43.47%±3.77%)(F=152.05,P<0.05)；第30d時交界區VEGF暘性細胞百分數(9.27%±2.08%)低于正常區(44.58%±3.34%)(F=274.1,P<0.052)。交界區的PCNA標記指數(PCNA labeling index，PCNA LI)(33.04%±4.31%)在第7d時低于正常區(65.15%±3.85%)，(F=242.46,P<0.05)；在14d時為(21.05%±1.96%)低于正常區(62.99%±3.34%)(F=413.52,P<0.05)；在30d時為(6.36%±0.51%)低于正常區(62.07%±18.07%)(F=729.59,P<0.05)。第7d時交界區的凋亡指數(apoptotic index，AI)(26.10%±4.54%)高于正常區(1.43%±0.35%)(F=216.22,P<0.05)。14d時交界區的凋亡指數AI(65.70%±1.14%)高于正常區(1.82%±0.31%)(F=1448.64,P<0.05)；30d時交界區的凋亡指數AI(82.02%±3.98%)高于正常區(2.52%±0.29%)(F=2244.33,P<0.05)。結論在本實驗設定參數及聲學劑量情況下, HIFU除瞭對照射區組織產生直接的殺滅作用外，還能抑製交界區的細胞增殖以及誘髮細胞凋亡的髮生。
목적：통과관찰고강도취초초성(high intensity focused ultrasound ，HIFU)대라서피하인효질류모형작용후혈관내피생장인자(vascular endothelial growth factor，VEGF)、증식세포핵항원(proliferating cell nuclear anti?gen，PCNA)단백적표체화세포적조망，탐토HIFU치료후대교계구급조사구세포증식화세포조망적영향。방법건립18지라서좌우배부피하효질류모형，용빈솔9.7MHz，초거4.5mm，성강2500W/cm2적HIFU련속조사20s。안조사후동물처사시간수궤분위7d조，14d조，30d조，매조각6지라서즉12개피하효질류조직；응용면역조화법검측HIFU작용후제7d、14d、30d조사구、교계구、정상구VEGF、PCNA단백적표체；응용원위말단표기법(TdT-mediated dUTP nick end labeling，TUNEL)검측조사구、교계구、정상구적조망세포。결과면역조화관찰도，각시각점교계구급정상구균유VEGF급PCNA표체병검측도조망염색양성적세포，조사구균무PCNA단백적표체급미검측도조망세포。제7d시교계구VEGF양성세포백분수(23.79%±3.11%)저우정상구(46.16%±2.43%)(F=110.03,P<0.05)；조사구VEGF양성세포백분수(10.94%±3.95%)저우정상구(46.16%±2.43%)(F=272.80， P<0.05)。제14d시교계구VEGF양성세포백분수(17.17%±2.89%)저우정상구(43.47%±3.77%)(F=152.05,P<0.05)；제30d시교계구VEGF양성세포백분수(9.27%±2.08%)저우정상구(44.58%±3.34%)(F=274.1,P<0.052)。교계구적PCNA표기지수(PCNA labeling index，PCNA LI)(33.04%±4.31%)재제7d시저우정상구(65.15%±3.85%)，(F=242.46,P<0.05)；재14d시위(21.05%±1.96%)저우정상구(62.99%±3.34%)(F=413.52,P<0.05)；재30d시위(6.36%±0.51%)저우정상구(62.07%±18.07%)(F=729.59,P<0.05)。제7d시교계구적조망지수(apoptotic index，AI)(26.10%±4.54%)고우정상구(1.43%±0.35%)(F=216.22,P<0.05)。14d시교계구적조망지수AI(65.70%±1.14%)고우정상구(1.82%±0.31%)(F=1448.64,P<0.05)；30d시교계구적조망지수AI(82.02%±3.98%)고우정상구(2.52%±0.29%)(F=2244.33,P<0.05)。결론재본실험설정삼수급성학제량정황하, HIFU제료대조사구조직산생직접적살멸작용외，환능억제교계구적세포증식이급유발세포조망적발생。
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/cm2). The 18 nude mice were randomly di?vided into 7 d group,14 d group and 30 d group according to sacrifice date. Immunohistochemical method, TdT-mediat?ed dUTP nick end labeling method were used to examine the expression of vascular endothelial growth factor and prolifer?ating 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 af?ter 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%±3.85%) in 7 d after ablation (F=242.46, P<0.05). The PCNA LI was lower in marginal zone than in normal zone (21.05%± 1.96%vs. 62.99%± 3.34%) in 14 d after ablation (F=413.52, P<0.05). The PCNA LI was lower in marginal zone than in normal zone (6.36%± 0.51% vs. 62.07%± 18.07%) in 30 d after ablation, (F=729.59, P<0.05) .The apoptotic index (AI) was higher in marginal zone than in normal zone (26.10%±4.54%vs. 1.43%±0.35%) in 7 d after ablation, (F=216.22, P<0.05). The apoptotic index(AI) was higher in marginal zone than in normal zone (65.70%± 1.14% vs. 1.82%± 0.31%) in 14d after ablation (F=1448.64, P<0.05). The apoptotic index (AI) was higher in marginal zone than in normal zone (82.02%± 3.98% vs. 2.52%± 0.29%) in 30d after ablation (F=2244.33, P<0.05). Conclusion The present study demonstrates that an extracorporeal HIFU with 9.7MHz transducer (the focal length of 4.5mm and fo?cal intensity 2500W/cm2) can completely ablate neurogliocytoma at exposure coverage and inhibit the proliferation of neurogliocytoma at marginal zone. Thus, HIFU may be a new and selective treatment for neurogliocytoma.
