【摘要】：目的:對比熒光分子探針四甲基羅丹明-葡聚糖(dextran-tetramethylrhodamine,DT)和熒光黃(lucifer yellow CH,LY)、磁性分子探針釓-二乙三胺五乙酸(gadolinium-diethylene triamine pentaacetic acid,Gd-DTPA)在多孔介質(zhì)中的動態(tài)擴散分布規(guī)律,篩選合適的熒光分子探針用于大鼠腦組織間隙(interstitial space,ISS)光學成像。方法:瓊脂糖凝膠分為DT組、LY組、Gd-DTPA組,分別導入相應(yīng)分子探針,應(yīng)用激光掃描共聚焦顯微鏡分別動態(tài)觀察DT、LY在瓊脂糖凝膠內(nèi)的擴散分布,與磁共振成像顯示的Gd-DTPA在瓊脂糖凝膠內(nèi)的動態(tài)擴散過程進行比較。LY分別導入18只大鼠尾狀核,對不同時間點離體腦切片進行熒光成像,其成像數(shù)據(jù)與相應(yīng)導入Gd-DTPA的大鼠的磁共振在體檢測成像數(shù)據(jù)進行對比。結(jié)果:DT、LY及Gd-DTPA在瓊脂糖凝膠中的擴散分布均表現(xiàn)為各向同性,平均擴散分布速率分別為:(0.07±0.02)×10-2mm2/s、(1.54±0.47)×10-2mm2/s、(1.45±0.50)×10-2mm2/s,DT與LY、DT與Gd-DTPA間差異均有統(tǒng)計學意義(ANOVA,F=367.15,P0.001;Post-Hoc LSD,P0.001),LY與Gd-DTPA間差異無統(tǒng)計學意義(Post-Hoc LSD,P=0.091)。重復測量方差分析比較分子探針的擴散分布面積隨時間變化的趨勢:DT與LY、DT與Gd-DTPA間的變化規(guī)律差異均有統(tǒng)計學意義(Bonferroni校正,α=0.0125,P0.001),LY與Gd-DTPA間差異無統(tǒng)計學意義(Bonferroni校正,α=0.0125,P=0.203)。LY與Gd-DTPA在大鼠尾狀核ISS內(nèi)分布隨時間變化呈各向異性,均表現(xiàn)為指向同側(cè)皮層區(qū)的單方向楔形擴散,平均擴散分布速率分別為(1.03±0.29)×10-3mm2/s和(0.81±0.27)×10-3mm2/s,t=0.759,P=0.490;信號衰減半衰期分別為(2.58±0.04)h和(2.46±0.10)h,t=2.025,P=0.113。LY與Gd-DTPA間擴散分布面積比率在0.5、1、2、3、7 h差異無統(tǒng)計學意義(t=2.249,P=0.088;t=2.582,P=0.061;t=1.966,P=0.121;t=0.132,P=0.674;t=0.032,P=0.976),在11 h差異有統(tǒng)計學意義(t=2.917,P=0.043)。結(jié)論:LY與Gd-DTPA在多孔介質(zhì)內(nèi)的擴散分布規(guī)律一致,是ISS熒光成像的適用分子探針,可用于腦ISS的微觀、宏觀、離體檢測。
[Abstract]:Objective: to compare the fluorescence molecular probes, tetramethylrhodamine-dextran (dextran-tetramethylrhodamine,DT) and fluorescent yellow (lucifer yellow CH,LY, and magnetic molecular probes, gadolinium-diethyltriamine pentaacetic acid (gadolinium-diethylene triamine pentaacetic acid,). The dynamic diffusion distribution of Gd-DTPA in porous media was studied. A suitable fluorescent molecular probe was selected for optical imaging of rat brain tissue gap (interstitial space,ISS). Methods: agarose gels were divided into three groups: DT group, LY group and Gd-DTPA group. The diffusion distribution of DT,LY in agarose gel was observed dynamically by laser scanning confocal microscope. The dynamic diffusion process of Gd-DTPA in agarose gel was compared with magnetic resonance imaging (MRI). LY was introduced into the caudate nucleus of 18 rats, and fluorescence imaging was performed on brain slices at different time points. The imaging data were compared with the magnetic resonance imaging (MRI) data of the rats introduced into Gd-DTPA. Results: the diffusion distribution of DT,LY and Gd-DTPA in agarose gel was isotropic, and the average diffusion rates were (0.07 鹵0.02) 脳 10 脳 10 ~ (2) mm ~ (2) vs (1.54 鹵0.47) 脳 10 ~ (2) mm ~ (2 / s), respectively. (1.45 鹵0.50) 脳 10 ~ (2) mm ~ (2), there was significant difference between DT and LY,DT and Gd-DTPA (ANOVA,F=367.15,P0.001;). There was no significant difference between Post-Hoc LSD,P0.001), LY and Gd-DTPA (Post-Hoc LSD,P=0.091). The variation trend of diffusion distribution area of molecular probe with time was compared by repeated measurement of variance analysis. The difference between DT and LY,DT and Gd-DTPA was statistically significant (Bonferroni correction, 偽 = 0.0125, P0.001), and the difference was statistically significant between the two groups (Bonferroni correction, 偽 = 0.0125, P0.001). There was no significant difference between LY and Gd-DTPA (Bonferroni correction, 偽 = 0.0125, P = 0.203). The distribution of LY and Gd-DTPA in the caudate nucleus ISS of rats showed anisotropy with time, which showed that the distribution of LY and Gd-DTPA was one-way wedge diffusion pointing to the ipsilateral cortex. The average diffusion rates were (1.03 鹵0.29) 脳 10-3mm2/s and (0.81 鹵0.27) 脳 10 脳 10 ~ (3) mm ~ (2 / s), t = 0.759, P = 0.490, respectively. The half-life of signal attenuation was (2.58 鹵0.04) h and (2.46 鹵0.10) h, t = 2.025, P = 0.113.LY / Gd-DTPA was 0.5,1,2,3 and 7 h, respectively, and there was no significant difference between them (t = 2.249, P = 0.088). T = 2.582, P = 0.061 / t / 1.966, P = 0.121 / t / 0.132, P = 0.674 / t / 0.032, P = 0.976), and there was a significant difference at 11 h (t = 2.917, P = 0.043). Conclusion: the diffusion distribution of LY and Gd-DTPA in porous media is consistent, and it is a suitable molecular probe for ISS fluorescence imaging. It can be used for microcosmic, macroscopical and in vitro detection of brain ISS.
【作者單位】： 大連大學環(huán)境與化學工程學院;北京大學醫(yī)藥衛(wèi)生分析中心;北京市磁共振成像設(shè)備與技術(shù)重點實驗室;北京大學第三醫(yī)院放射科;北京大學第三醫(yī)院神經(jīng)科;北京大學第三醫(yī)院臨床流行病學研究中心;
【基金】：國家自然科學基金(61450004;91330103) 北京市科技專項(z141107004414031) 高等學校博士學科點專項科研基金(20130001130013)資助~~
【分類號】：R445.2

【參考文獻】

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