5-氨基乙酰丙酸介导鼠脑C6胶质瘤荧光特性的研究  

The Study of Fluorescence Character in C6 Cell and Rat C6 Glioma Model after Administration of 5-aminolevulinic Acid

作　　者：滕雷[1] 鞠东辉[1] 单强[1] 李一[1] 于洪伟[1] 韩大勇[1] 汪立刚[1] 赵世光[1] 

机构地区：[1]哈尔滨医科大学附属第一临床医学院神经外科,黑龙江哈尔滨150001

出　　处：《中国神经肿瘤杂志》2008年第1期25-29,共5页Chinese Journal of Neuro-Oncology

基　　金：哈尔滨市科技攻关计划项目(No.2007AA3CS083-2)

摘　　要：背景与目的:5-氨基乙酰丙酸(5-AlA)荧光引导手术切除神经胶质瘤的基础研究一直较为薄弱,本文旨在探讨5-AlA诱导C6胶质瘤内生性卟啉荧光随时间的动力学变化及卟啉在C6胶质瘤和正常鼠脑中的分布。方法:将C6细胞与5-ALA共培养后,使用流式细胞仪检测细胞内原卟啉Ⅸ(Protoporphyrin Ⅸ,Pp Ⅸ)含量。经股静脉向C6/Wistar大鼠脑胶质瘤模型体内注入5-ALA后,使用荧光显微镜和激光扫描共聚焦显微镜观察C6 胶质瘤和正常鼠脑组织中卟啉荧光的分布。用鼠的冻伤皮质切片分析血脑屏障的破坏在卟啉产生中的影响。结果:体外C6细胞在加入5-ALA后1h检测到卟啉生成,主要集中在5-15h,11h达到高峰。C6胶质瘤切片荧光主要集中在注入5-ALA后2～10h,6h表现出最强荧光。肿瘤整体荧光呈斑片状分布,边界欠清楚。给药后对侧脑组织未检测到低水平荧光。给药后5h在皮质冻伤切片中检测到微量荧光。无瘤鼠脑组织在注入5-ALA后没有检测到荧光。结论:5-AlA能诱导卟啉在C6细胞中积聚,其荧光表现是一个动态过程。5-ALA介导胶质瘤内生性卟啉蓄积与血脑屏障具有密切关系。肿瘤与正常脑组织具有不同荧光表现强度,可以为临床选择性应用5-ALA荧光引导胶质瘤切除术提供有益的指导作用。BACKGROUND ＆ OBJECTIVE：In this work,the photodynamic effects dependent time of 5-ALA in C6 cell and rat C6 glioma model were comparatively studied. METHODS ：C6 tumour cell cultures were incubated with 5-ALA in vitro.Fluorescence was measured with a flow cytometry in cell cultures.Rat C6 glioma model was established by implanting C6 glioblastoma cells into rat brain by means of stereotactic injection.The fuorescence distribution of endogenous porphyrins was studied in brain tissue sections with fuorescence microscope and confocal laser scanning microscope.The role of blood-brain barrier disruption on porphyrin production was studied with a cryo-lesion of the cortex. RESULTS：Porphyrins were detected in vitro from 1 hour and in vivo from 2 to 10 hours,by 20 hours porphyrin fuorescence had almost disappeared.Fluorescence in the C6 tumours was patchy,high fuorescence was detected levels between 2 and 10 hours after 5-ALA administration. Low fuorescence was measured 6 hours after 5-ALA administration in cryo-lesions model.The normal brain was not stained in contral rat. CONCLUSIONS ：5-ALA can induce endogenous porphyrin generation in C6 eel] in vivo and in vitro,its fluomseenee ehange was dynamie process dependent time.Endogenous porphyrin generation is linked to blood-brain barrier disruption.The extent of fluomseenee in site, sueh as tumour and normal brain tissue,was defferent,easily identied.Henee, fluoreseenee image guided bain tumor resection is thought to assist neurosurgeons to re sect malignant glioma.

关 键 词：5-氨基乙酰丙酸 内生性卟啉 血脑屏障 荧光检测 

分 类 号：R739.41[医药卫生—肿瘤]