荧光及磁示踪法观测脑组织液的引流分区特征

发布时间：2018-06-01 15:45

  本文选题：脑 + 分子探针技术　； 参考：《北京大学学报(医学版)》2017年02期

【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±2.88)mm~2、(19.00±5.21)mm~2vs.(15.75±2.29)mm~2、(25.32±5.49)mm~2vs.(22.33±3.25)mm~2、(17.34±5.31)mm~2vs.(15.92±2.75)mm~2、(7.67±6.19)mm~2vs.(5.00±1.89)mm~2,对其在每层两组间扩散面积进行t检验,差异均无统计学意义(t=0.753,P=0.493;t=1.875,P=0.134;t=0.990,P=0.378;t=0.810,P=0.464;t=0.413,P=0.701;t=0.716,P=0.514);Sn亚组LY与Gd-DTPA导入1 h后,比较1~6层LY与Gd-DTPA扩散面积为:(6.78±4.56)mm~2vs.(4.75±2.00)mm~2、(12.65±5.04)mm~2vs.(10.44±1.13)mm~2、(19.51±6.54)mm~2vs.(17.55±0.30)mm~2、(28.72±5.45)mm~2vs.(24.48±1.32)mm~2、(21.34±4.42)mm~2vs.(17.72±0.25)mm~2、(13.00±5.46)mm~2vs.(12.00±2.88)mm~2,对其在每层间扩散面积进行t检验,扩散面面积的差异均无统计学意义(t=0.705,P=0.519;t=0.743,P=0.499;t=0.517,P=0.656;t=1.310,P=0.260;t=1.416,P=0.292;t=0.281,P=0.793),但LY扩散面积略大于Gd-DTPA。结论:荧光法证实了磁示踪法发现的ISF在ISS内的分区引流特征,并且可为磁示踪法提供离体验证的技术与方法。荧光ISS成像法具有更高的对比度和分辨率,得到了更加精细的ISF引流分区区域。
[Abstract]:Aim: to compare the results of molecular diffusion and mass flow motion in interstitial space ISSs (interstitial space ISSs) with two magnetic probes, and to study the regional drainage characteristics of interstitial fluid flow (ISF) in ISS. Methods Thirty six Sprague-Dawley rats were randomly divided into three subgroups: fluorescent inspection group (n = 18) and magnetic tracer group (n = 18). Each group was further divided into three subgroups: caudate nucleusus caudate, thalamus and substantia nigragragragrausin, with 6 rats in each subgroup. Fluorescence inspection group: referring to the stereotaxic map of the brain, the coronal globus pallidus was selected as the central layer, the CNT and Sn were used as the target for puncture localization, and 2 渭 L 10 mmol/L fluorescent lucifer low y) was introduced into the central position of the corresponding brain region, respectively. The rats were fixed with 4% (volume fraction) of paraformaldehyde for 2 h and Sn 1 h respectively. The brain was placed in the brain slice mould and sliced backward along the optic chiasma. The coronal section of the needle was taken as the central layer. There were 1 slice, 3 pieces before and 2 pieces after, each slice was about 1 mm thick, 6 pieces were to be detected. Laser scanning confocal microscope (LSCM) was used to collect the in vitro slice and the area of LY diffusion was measured. In the magnetic tracer group, CNT and Sn were also used as the target point for the puncture localization of 2 渭 L 10 mmol/L Gadolinium-diethylene triamine pentaacetic acidmite Gd-DTPAA. The diffusion of the tracer in the rat brain was dynamically monitored by magnetic resonance imaging technique, and the diffusion area of Gd-DTPA was measured by Radiant software. 缁撴灉:LY涓嶨d-DTPA鍦–n,T鍜孲n鐨処SS鍐呮墿鏁ｅ尯鍩熷悇涓嶇浉鍚，

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