外源化合物跨血脑屏障转运动力学模型的建立与应用

发布时间：2018-02-02 22:44

本文关键词： 原位脑灌流技术血脑屏障转运动力学肼一甲基肼噻诺啡　出处：《中国人民解放军军事医学科学院》2005年硕士论文　论文类型：学位论文

【摘要】：原位脑灌流技术是研究物质跨BBB转运机制的一种快速、敏感、定量、可控的方法,在药物或毒物动力学上有广泛的应用。它可简便地描绘出物质入脑的动力学变化,结合血管清除方法可以区分真正的跨膜转运入脑和单纯的内皮细胞积聚,可研究物质的跨BBB转运机制。本研究应用大鼠原位脑灌流技术,结合血管清除方法和多点吸收数据分析,建立了外源化合物跨BBB转运的动力学模型。大鼠原位脑灌流系统主要由液体输送系统及其附属设备组成,各部分均经过严格的挑选与调试;并在硬件构建完成后对该技术的操作和各种条件进行了优化。经实验验证原位脑灌流是表达血脑转运特征的一种可靠技术,其操作比较简便,具有活体的真实性,且方法可靠,条件可控,结果稳定。血管清除实验可以有效区分真正入脑实质的转运和血管组织的内吞与简单结合。多点吸收数据分析方法能够表征外源化合物进入脑组织的动力学特征。重新确证了肼和MMH在脑组织样品中的对-二甲氨基苯甲醛(p-DMBA)比色测定方法;新建立了噻诺啡的以液-液萃取为生物样品前处理的HPLC-UV定量分析方法,并从特异性、灵敏度、线性范围、精密度、准确度以及样品的稳定性进行了方法学的确证。这三种化合物在脑组织样品中的测定方法均符合动力学实验要求,能够满足各自跨BBB转运动力学研究的需要。采用大鼠原位脑灌流方法,结合血管清除方法以及多点吸收分析的动力学模型研究了肼、MMH和噻诺啡三种外源化合物跨BBB转运的动力学特征,结果显示三者均可跨过BBB进入脑实质,入脑浓度随着灌流浓度和灌流时间的增加而呈上升趋势;三者在各自灌流浓度下跨BBB转运的速度常数K_(in)依次为0.01、0.02和
[Abstract]:In situ cerebral perfusion is a rapid, sensitive, quantitative and controllable method to study the transport mechanism of substance across BBB. It is widely used in the pharmacokinetics of drugs or poisons. It can easily describe the dynamic changes of substance into the brain and can distinguish the true transmembrane transport into the brain and the accumulation of endothelial cells in combination with vascular clearance. In this study, we used in situ brain perfusion technique, combined with vascular clearance method and multi-point absorption data analysis. A kinetic model for the transport of exogenous compounds across BBB was established. The in-situ cerebral perfusion system in rats was mainly composed of liquid delivery system and its auxiliary equipment. All parts were selected and adjusted strictly. After the completion of the hardware construction, the operation and various conditions of the technique were optimized. It is proved by experiments that in situ brain perfusion is a reliable technique to express the characteristics of blood brain transport, and its operation is relatively simple. The method is reliable and the condition is controllable. The results are stable. Vascular clearance experiments can effectively distinguish the transport into brain parenchyma and the endocytosis and simple combination of vascular tissue. Multi-point absorption data analysis method can characterize the dynamic characteristics of exogenous compounds entering brain tissue. . The colorimetric method of p-dimethylaminobenzaldehyde (p-DMBA) for determination of hydrazine and MMH in brain tissue samples was reconfirmed. A new HPLC-UV method for quantitative analysis of tienorphine with liquid-liquid extraction as biological sample was established, and the specificity, sensitivity, linear range and precision were analyzed. The accuracy and stability of the samples were confirmed by the methodology. The determination of these three compounds in brain tissue samples met the requirements of kinetic experiments. It can meet the needs of the study on the transport kinetics of BBB. The kinetic characteristics of the transport of three exogenous compounds of hydrazine and tienorphine through BBB were studied by in situ cerebral perfusion in rats combined with vascular clearance and multipoint absorption analysis. The results showed that all three crossed BBB into brain parenchyma, and the brain concentration increased with the increase of perfusion concentration and perfusion time. The velocity constant of BBB transport among the three groups under their respective irrigation concentration is 0.01g / 0.02 and 0.01 / 0. 02, respectively.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R363

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