磁共振弥散成像评价血管内皮生长因子质粒治疗犬脑梗死的有效性
자공진미산성상평개혈관내피생장인자질립치료견뇌경사적유효성
Evaluating the therapeutic efficacy of vascular endothelium growth factor plasmid in treating dog cerebral infarction with diffusion-weighted magnetic resonance imaging
  • 万方数据
    中国临床康复 중국림상강복
    CHINESE JOURNAL OF CLINICAL REHABILITATION
    2005年 25期 210-213 ,共4页

    刘怀军%武柏林%汪国石%王藏海%王立新%玉鉴%屈长强%池琛%黄渤源

    류부군%무백림%왕국석%왕장해%왕립신%옥감%굴장강%지침%황발원

    脑梗塞%磁共振成像%疾病模型%动物%内皮生长因子 腦梗塞%磁共振成像%疾病模型%動物%內皮生長因子
    뇌경새%자공진성상%질병모형%동물%내피생장인자
    背景:在脑梗死后恢复过程中血管内皮细胞生长因子是一种强效的内皮细胞有丝分裂原和促血管因子,应用磁共振弥散成像是否可以记录到这种治疗作用.目的:用磁共振弥散加权成像追踪观察血管内皮细胞生长因子质粒治疗局灶性脑梗死家犬动物模型的效果,并将形态学检查和免疫组化染色结果相对照.设计:完全随机对照,盲法评估,方差设计,Person相关分析,2周跟踪观察.单位:河北医科大学第二医院医学影像科.材料:实验于2001-04/2002-03在河北医科大学第二医院影像科实验室完成,选择18只体质量10～15 kg的健康成年家犬.随机分为2组,对照组9只和实验组9只.方法:所有动物股动脉插管,颈内动脉推注栓子到达大脑中动脉制成脑梗死动物模型.对照组分别于术后24h,1周,2周处死,每个时间点3只,实验组于术后1周处死4只,术后2周处死5只.实验组采用微量注射器注入含pcD2/hVEGF121质粒的溶液0.5 mL(500～600 μg).对照组给予等容积生理盐水.手术后磁共振扫描,1次/h,共4次.扫描序列为T1WI,T2WI,3D-TOFMRA,DWI,CET1WI.术后24 h,3 d,1周、2周重复上述序列扫描.动物处死后以MR图像为依据选择病变部位的细胞形态观察采用苏木精-伊红染色、微血管记数采用CD34免疫组化染色,血管内皮细胞生长因子免疫阳性细胞数采用血管内皮细胞生长因子免疫组化染色.计算表观分布系数,方差分析法分析不同时间不同处理组之间的差异.计数每高倍视野下的毛细血管数,计数血管内皮细胞生长因子免疫阳性细胞数.将MR扫描结果与免疫组化结果对照,分析MR信号改变同免疫组化结果之间是否存在相关关系.主要观察指标:①各组动物术后24h,1周,2周时脑梗死区及对侧相应区微血管计数及血管内皮细胞生长因子免疫阳性细胞计数.②各组动物脑磁共振成像检查结果.结果:18只家犬均进入结果分析.①术后1 h磁共振弥散加权成像扫描可见到梗死区明显高信号,且随时间的延长强度逐渐增高.②表观分布系数值术后三四个小时下降到(5.611.39)mm2/s,相对于对侧半球(9.85±2.04)mm2/s下降了约43%,2周时达(9.83±1.11)mm2/s,仍然略低于正常.③后续磁共振扫描显示表观分布系数比率由超急性期的下降开始呈上升趋势,对照组升高更显著,2周时组间出现显著差异(P=0.032,0.006).④2周时实验组患侧微血管计数免疫阳性细胞显著高于对照组[(28.80±3.29)个/视野,(20.70±4.47)个/视野,(P＜0.01)].⑤实验组患侧1,2周时血管内皮生长因子免疫阳性细胞数显著高于对照组[(64.20±9.40)个/视野,(51.90±5.74)个/视野;(72.70±6.98)个/视野,(58.40±6.35)个/视野,(P＜0.01)].⑥将磁共振检查结果同免疫组化结果对照进行相关分析,发现表观分布系数比率变化与微血管计数之间存在显著的相关关系,Pearson相关系数为0.679,P＜0.01.微血管计数和血管内皮生长因子阳性细胞数之间有显著相关关系(r=0.668,P＜0.01).结论:形态学观察和免疫组织化学证实采用血管内皮细胞生长因子质粒基因治疗后,随时间进展局部微血管明显增多,血管内皮细胞生长因子蛋白含量上升.同时表观分布系数的变化与血管内皮细胞生长因子免疫阳性细胞计数及微血管计数的变化呈显著的正相关关系.
    배경:재뇌경사후회복과정중혈관내피세포생장인자시일충강효적내피세포유사분렬원화촉혈관인자,응용자공진미산성상시부가이기록도저충치료작용.목적:용자공진미산가권성상추종관찰혈관내피세포생장인자질립치료국조성뇌경사가견동물모형적효과,병장형태학검사화면역조화염색결과상대조.설계:완전수궤대조,맹법평고,방차설계,Person상관분석,2주근종관찰.단위:하북의과대학제이의원의학영상과.재료:실험우2001-04/2002-03재하북의과대학제이의원영상과실험실완성,선택18지체질량10～15 kg적건강성년가견.수궤분위2조,대조조9지화실험조9지.방법:소유동물고동맥삽관,경내동맥추주전자도체대뇌중동맥제성뇌경사동물모형.대조조분별우술후24h,1주,2주처사,매개시간점3지,실험조우술후1주처사4지,술후2주처사5지.실험조채용미량주사기주입함pcD2/hVEGF121질립적용액0.5 mL(500～600 μg).대조조급여등용적생리염수.수술후자공진소묘,1차/h,공4차.소묘서렬위T1WI,T2WI,3D-TOFMRA,DWI,CET1WI.술후24 h,3 d,1주、2주중복상술서렬소묘.동물처사후이MR도상위의거선택병변부위적세포형태관찰채용소목정-이홍염색、미혈관기수채용CD34면역조화염색,혈관내피세포생장인자면역양성세포수채용혈관내피세포생장인자면역조화염색.계산표관분포계수,방차분석법분석불동시간불동처리조지간적차이.계수매고배시야하적모세혈관수,계수혈관내피세포생장인자면역양성세포수.장MR소묘결과여면역조화결과대조,분석MR신호개변동면역조화결과지간시부존재상관관계.주요관찰지표:①각조동물술후24h,1주,2주시뇌경사구급대측상응구미혈관계수급혈관내피세포생장인자면역양성세포계수.②각조동물뇌자공진성상검사결과.결과:18지가견균진입결과분석.①술후1 h자공진미산가권성상소묘가견도경사구명현고신호,차수시간적연장강도축점증고.②표관분포계수치술후삼사개소시하강도(5.611.39)mm2/s,상대우대측반구(9.85±2.04)mm2/s하강료약43%,2주시체(9.83±1.11)mm2/s,잉연략저우정상.③후속자공진소묘현시표관분포계수비솔유초급성기적하강개시정상승추세,대조조승고경현저,2주시조간출현현저차이(P=0.032,0.006).④2주시실험조환측미혈관계수면역양성세포현저고우대조조[(28.80±3.29)개/시야,(20.70±4.47)개/시야,(P＜0.01)].⑤실험조환측1,2주시혈관내피생장인자면역양성세포수현저고우대조조[(64.20±9.40)개/시야,(51.90±5.74)개/시야;(72.70±6.98)개/시야,(58.40±6.35)개/시야,(P＜0.01)].⑥장자공진검사결과동면역조화결과대조진행상관분석,발현표관분포계수비솔변화여미혈관계수지간존재현저적상관관계,Pearson상관계수위0.679,P＜0.01.미혈관계수화혈관내피생장인자양성세포수지간유현저상관관계(r=0.668,P＜0.01).결론:형태학관찰화면역조직화학증실채용혈관내피세포생장인자질립기인치료후,수시간진전국부미혈관명현증다,혈관내피세포생장인자단백함량상승.동시표관분포계수적변화여혈관내피세포생장인자면역양성세포계수급미혈관계수적변화정현저적정상관관계.
    BACKGROUND: Vascular endothelium growth factor (VEGF) is an endothelium mitogen and angiogenic factor with strong potential during recovery from cerebral infarction (CI). Can such therapeutic effect be detected with magnetic resonance diffusion imaging?OBJECTIVE: To study the therapeutic efficacy of VEGF plasmid in treating focal cerebral infarction in a dog experimental model with the aid of diffusion- and hemodynamic-weighted magnetic resonance imaging (MRI),with the morphological results compared with those of immunohistochemical examination.DESIGN: Completely randomized controlled, double blind evaluation,analysis of variance, Pearson correlation analysis, follow-up for 2 weeks.SETTING: Department of Medical Iconography, the Second Affiliated Hospital of Hebei Medical University.MATERIALS: This study was carried out at the Department of Medical Iconography, the Second Affiliated Hospital of Hebei Medical University,between April 2001 and March 2002. Totally 18 healthy adult dogs weighing 10-15 kg were randomly divided into control group and experiment group with half in each.METHODS: All dogs were subjected to femoral intubation and then made into CI model by the occlusion of middle cerebral artery with an embolus injected through the internal carotid artery. Dogs in control group were put to death at postoperative 24 hours, 1 week and 2 weeks with three at each time point, while four dogs in experiment group were put to death at postoperative 1 week and five at 2 weeks. Dogs in experiment group received microinjection of 0.5 mL fluid containing pcD2/hVEGF121 (500-600 μg)instantly after operation, which was replaced with physical saline of the same volume at the same time point in control group. Then they were subjected to MRI scanning once an hour for 4 times, with the sequence of T1WI, T2WI, 3D-TOFMRA, DWI and CET1WI, which was repeated at postoperative 24 hours, 3 days, 1 week and 2 weeks. Based on the MR images, pathological focuses were selected for morphological observation of cells with the aid of HE staining, and CD34 IHC staining was used for counting micrangium, as well as VEGF staining for VEGF positive cells.Then the apparent distribution coefficient (ADC) was calculated, and the differences between different time points and groups were analyzed by analysis of variance. The number of capillaries and VEGF positive cells of each high-power field was counted, with the results compared with those of MR scanning so as to explore the correlation between MR signal changes and IHC results.MAIN OUTCOME MEASURES: ① The number of capillaries and VEGF positive cells in each high-power field was counted at postoperative 24 hours, 1 week and 2 weeks; ② MR images of each group.RESULTS: Data of the 18 dogs entered the final analysis. ① Diffusionweighted imaging (DWI) showed higher signals at infarctional region at postoperative 1 hour, which became strengthened as time went by. ②ADC decreased to (5.61 ±1.39) mm2/s at postoperative 3-4 hours, about 43% lower than that of the opposite hemisphere [(9.85±2.04) mm2/s]. It resumed to (9.83±1.11) mm2/s, but was still lower than the normal level.③ The subsequent MR scanning proved that ADC ratio presented an increasing tendency in contrast with the decreasing tendency at super-acute stage. The increment was even more marked in control group and the difference was significant at postoperative 2 weeks (P=0.032, 0.006). ④ The number of capillary positive cells on the affected side in experiment group was significantly higher than that in control group at postoperative 2 weeks [(28.80±3.29)/field, (20.70±4.47)/field, (P ＜ 0.01)]. ⑤ The number of VEGF positive cells on the affected side in experiment group was significantly higher than that in control group at postoperative 1 and 2weeks [(64.20±9.40)/field, (51.90±5.74)/filed; (72.70±6.98)/filed,(58.40±6.35)/field, (P ＜ 0.01)].⑥ The results of MR scanning and IHC were subjected to correlation analysis and revealed that ADC ratio was closely correlated with the number of capillary positive cells, with Pearson correlation coefficient being 0.679 (P ＜ 0.01). Moreover, the number of capillaries and the number of VEGF positive cells were significantly correlated (r=0.668, P ＜ 0.01).CONCLUSION: Morphological observation and IHC revealed that both the local capillaries and VEGF protein content increased markedly in timedependant manner due to VEGF plasmid gene therapy. Meanwhile, the change of ADC ratio was found to be closely correlated with the number of VEGF positive cells and the number of capillaries.
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