荧光及磁示踪法观测脑组织液的引流分区特征  

Drainage characteristic of the brain interstitial fluid detected by using fluorescence and magnetic tracer method

作　　者：赵越[1,2,3] 李昀倩[2,3] 李怀业[1,2,3] 李玉亮[4] 刘兰祥[5] 袁兰[2] 张殊佳[1] 韩鸿宾[3,6] 

机构地区：[1]大连大学环境与化学工程学院,辽宁大连116622 [2]北京大学医药卫生分析中心,北京100191 [3]北京市磁共振成像设备与技术重点实验室,北京100191 [4]山东大学第二医院介入科,济南250033 [5]秦皇岛市第一医院磁共振室,河北秦皇岛066000 [6]北京大学第三医院放射科,北京100191

出　　处：《北京大学学报（医学版）》2017年第2期303-309,共7页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(91330103;81471633);中国博士后科学基金(2015M570901);博士点优先发展领域基金(20130001130013);北京大学第三医院院重点项目(BYSY201301)资助~~

摘　　要：目的:对比光、磁两种探针示踪显示脑组织间隙(interstitial space,ISS)内分子扩散与团流运动的结果,研究脑组织液(interstitial fluid,ISF)在ISS内的分区引流特征。方法:36只SD大鼠随机分为荧光检查组(18只)和磁示踪组(18只),每组再随机分为尾状核(caudate nucleus,Cn)、丘脑(thalamus,T)和黑质(substantia nigra,Sn)3个亚组,每亚组6只。荧光检查组:参考脑立体定位图谱,选取冠状位苍白球为中心层面,以Cn、T和Sn为靶点进行穿刺定位,分别导入2μL 10 mmol/L荧光黄(lucifer yellow,LY)于相应脑区的中心位置,于Cn 3 h、T 2 h和Sn 1 h对大鼠用4%(体积分数)多聚甲醛进行心脏灌注固定后,将取出的鼠脑置于脑切片模具中,沿视交叉向后切片,以进针位点所在的冠状切片为中心层面,计为1片,前取3片,后取2片,每片厚约1 mm,共6片待检测,应用激光扫描共聚焦显微镜(laser scanning confocal microscope,LSCM)对离体切片进行采集,测量离体切片LY扩散的面积;磁示踪组大鼠也以Cn、T和Sn为靶点进行穿刺定位导入2μL 10 mmol/L钆喷酸葡胺(gadolinium-diethylene triamine pentaacetic acidm,Gd-DTPA),应用磁共振(magnetic resonance imaging,MRI)动态监测示踪剂在大鼠脑内的扩散,并利用Radiant软件测量Gd-DTPA的扩散面积。结果:LY与Gd-DTPA在Cn、T和Sn的ISS内扩散区域各不相同,Cn亚组LY与Gd-DTPA导入3 h后,比较1~6层LY与Gd-DTPA扩散面积为:(10.95±4.27)mm^2vs.(8.33±2.25)mm^2、(18.16±4.74)mm^2vs.(16.42±2.88)mm^2、(24.57±3.65)mm^2vs.(20.75±2.29)mm^2、(34.81±3.32)mm^2vs.(28.88±1.51)mm^2、(30.53±3.12)mm^2vs.(20.92±2.75)mm^2、(12.15±4.92)mm^2vs.(10.00±1.89)mm^2,对其在每层两组间扩散面积进行t检验,扩散面积差异均无统计学意义(t=0.940,P=0.400;t=0.546,P=0.614;t=1.534,P=0.200;t=2.809,P=0.480;t=2.693,P=0.055;t=0.707,P=0.518);T亚组中LY与Gd-DTPA导入2 h后,比较1~6层LY与Gd-DTPA扩散面积为:(5.56±4.61)mm^2vs.(3.33±2.25)mm^2、(16.21±3.36)mm^2vs.(11.42±2.8Objective：Compare the results of molecular diffusion and mass flow in the interstitial space（ISS） displayed by using optical and magnetic probes and study partitioned drainage of the brain interstitial fluid （ISF）.Methods： In the study,36 male SD rats were randomly divided into fluorescent inspection group （18）,magnetic tracer group （18）.Then they were divided equally into caudate nucleus （Cn）,thalamus （T） and substantia nigra （Sn） subgroup,6 rats in each subgroup.Referencing the brain stereotaxic atlas,the coronal globus pallidus as center level,Cn,T or Sn were acted as puncture positioning target.A 10 μL microsyringe was stereotaxically positioned and the lucifer yellow （LY） solution of 2 μL 10 mmol/L was infused into centric position.The coronary slices undergo cardiac perfusion and fix respectively in time point Cn 3 h,T 2 h and Sn 1 h.The rat brain was placed in rat stainless steel brain matrices and cut backward along visual intersection.The injection point of coronal slice as the center level,take 3 slices in front of the center level and 2 slices behind of it.1 mm for each slice and 6 slices in total.Then slices were detected by laser scanning confocal microscope （LSCM）.Simultaneous,in the same coordinate brain regions of another three groups,a gadolinium-diethylene triamine pentaacetic acidm （Gd-DTPA） solution of 2 μL 10 mmol/L was infused into different injection and detected by MRI tracer-based method.Then the Radiant can be used to measure distribution area of Gd-DTPA.Results： LY and Gd-DTPA have different distribution regions in Cn,T and Sn.After LY and Gd-DTPA were introduced into the Cn subgroup 3 h,compare the 1 to 6 levels distribution area of LY and Gd-DTPA as follows： （10.95±4.27） mm2 vs.（8.33±2.25） mm2,（18.16±4.74） mm2 vs.（16.42±2.88） mm2,（24.57±3.65） mm2 vs.（20.75±2.29） mm2,（34.81±3.32） mm2 vs.（28.88±1.51） mm2,（30.53±3.12） mm2 vs.（20.92±2.75） mm2,（12.15±4.92） mm2 vs.（10.00±1.89） mm2.

关 键 词：脑 分子探针技术 显微镜检查 共焦 磁共振成像 

分 类 号：R445.2[医药卫生—影像医学与核医学]