帕金森病6-OHDA模型大鼠左旋多巴血药浓度与纹状体细胞外液氨基酸类递质结合研究的方法  

The method of simultaneous measurement of the plasma concentration of L-DOPA and extracellular amino acids in striatum of the Parkinson's disease rats induced by 6-OHDA

作　　者：吴兆恩[1] 王丹巧[1] 李鹏[1] 孙晓芳[1] 王巍[2] 

机构地区：[1]中国中医科学院医学实验中心,北京100700 [2]中国中医科学院西苑医院,北京100091

出　　处：《中国药理学通报》2010年第10期1395-1399,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30772780);中国中医科学院基本科研业务费自主选题资助项目(NoZZ2006037)

摘　　要：目的建立对清醒自由活动的帕金森病模型大鼠进行左旋多巴(L-DOPA)血药浓度与纹状体细胞外液氨基酸类神经递质水平同步动态监测、结合研究的方法。方法 SD大鼠,脑内注射6-羟基多巴胺(6-OHDA)造模,血、脑双位点微透析采样、高效液相-荧光方法(HPLC-FLD)检测相关物质的浓度。结果大鼠腹腔给药吸收迅速,L-DOPA药时曲线符合一室模型;美多巴高剂量组的AUC、Cmax明显高于美多巴低剂量组(P<0.05或P<0.01)。与对照组相比,模型组大鼠纹状体细胞外液谷氨酸(Glu)水平明显升高,γ-氨基丁酸(GABA)水平明显降低;美多巴投药改善了6-OHDA导致的脑内氨基酸递质紊乱(P<0.05或P<0.01)。相同时间点血中L-DOPA浓度与脑内Glu/GABA值进行回归分析,r2值为0.7950。结论采用血、脑双位点微透析采样、HPLC-FLD检测技术,实现了对清醒自由活动大鼠L-DOPA血药浓度与纹状体细胞外液药效指标同步、动态的观察,提供了反映L-DOPA血药浓度-纹状体效应-时间三维关系的直接依据。Aim To establish the method of pharmacokineticpharmacodynamic（PK-PD） for levodopa（L-DOPA） by simultaneously monitoring the plasma concentration of L-DOPA and the extracellular amino acids level in striatum in the freely moving rats of Parkinson＇s disease.Methods The model of Parkinson＇s disease were induced by injecting 6-hydrodopamine（6-OHDA） into striatum of SD rats.Blood-brain microdialysis technique were used for simultaneously sampling,and high performance liquid chromatography-fluorescence detector（HPLC-FLD） was used to detect the concentration of related substances.Results Drugswere absorbed rapidly after administration through intraperitoneal injection and the plasma concentration-time profiles of L-DOPA were described by a one compartment model.AUC,C max,was higher in high madopar groups than that in low madopar ones（P 0.05 or P0.01） .Compared with control groups,the levels of Glu increased whileγ-aminobutyrate（GABA） decreased significantly after injecting 6-hydrodopamine（6-OHDA） into striatum. Madopar improved the disorder of amino acids induced by 6-OHDA（P0.05 or P0.01） .The regression analysis of the concentration of L-DOPA in plasma and striatum Glu/GABA in the same time point was made and the relative coefficient（r2） was 0.7950.Conclusion In the study,the blood-brain microdialysis technique combined with HPLC-FLD was established to detect the extracellular pharmacodynamic in striatum and the plasma concentration of L-DOPA in freely moving rats simultaneously,by which a three-dimensional relationship of L-DOPA among blood concentration,striatum effects and time was realized.

关 键 词：脑内微透析 血液微透析 左旋多巴 药动-药效学 谷氨酸 γ-氨基丁酸 6-羟基多巴胺 

分 类 号：R332[医药卫生—人体生理学] R322.81[医药卫生—基础医学]