可控释左旋多巴和苄丝肼微球减少帕金森病大鼠异动症的发生  

An experiment on controlled release levodopa/benserazide-loaded microspheres in treatment of levodopa-induced dyskinesia of rats with Parkinson＇ s disease

作　　者：杨新新[1] 任甜甜[2] 吴娜[1] 宋璐[1] 袁伟恩[2] 刘振国[1] 

机构地区：[1]上海交通大学医学院附属新华医院神经内科,200092 [2]上海交通大学药学院

出　　处：《中华神经科杂志》2011年第12期820-825,共6页Chinese Journal of Neurology

基　　金：国家自然科学基金资助项目（81071025）;上海市科委基础研究重点资助项目（09JC1411000）;上海市教委科研创新重点资助项目（10ZZ72）;上海市卫生局青年项目

摘　　要：目的观察聚乳酸-羟基乙酸共聚物（PLGA）包裹的可释放左旋多巴和苄丝肼的微球对帕金森病（PD）大鼠运动症状及异动症发生的影响并探讨其机制。方法PLGA包裹左旋多巴及苄丝肼制作微球，高效液相法测定微球在体内释放出的左旋多巴和苄丝肼的浓度，6-羟基多巴胺（6-OHDA）注射制作PD大鼠模型，制模成功的PD大鼠随机分成PD组、左旋多巴组、微球组（每组12只），另设溶剂注射假手术组（n=12）。左旋多巴组和微球组大鼠分别接受左旋多巴和苄丝肼（左旋多巴12mg／kg，苄丝肼15mg／kg）或等剂量微球皮下注射，在治疗的第1、4、7、10、14天行大鼠前肢功能测定，治疗2周后行大鼠异常不自主运动（AIM）评分，免疫组织化学及Westernblot法检测纹状体区磷酸化的多巴胺和环磷腺苷调节的磷酸化蛋白-32（DARPP-32）（Thr34）和AFosB水平。结果体内释放实验表明第7天时左旋多巴和苄丝肼释放量分别达76．2％和83．6％。微球处理组大鼠在治疗的第10天和第14天时前肢跨步数分别为5．8±1．6和5．2±1．5，比左旋多巴组（2．4±1．1、1．2±0．5）明显增加（t=4．12，5．43，均P〈0．01）。微球处理组大鼠第14天AIM评分[（16．0±2．1）分]较左旋多巴处理组[（26．0±3．2）分]显著下降，差异有统计学意义（t=6．59，P〈0．01）。免疫组织化学显示微球处理组大鼠纹状体磷酸化DARPP-32水平[（3．7±1．3）×10^4]较左旋多巴处理组[（7．9±2．2）×10^4]明显降低（t：2．95，P〈0．05）。Westernblot结果显示微球处理组大鼠磷酸化DARPP-32和AFosB水平分别为119．4％±11．3％和149．3％±12．3％，较左旋多巴组（184．8％±13．7％和300．4％±14．2％）显著下降（t=4．12、2．91，均P〈0．05）。结论微球皮下注射可以改善PD大鼠的运动症状，同时可以减少PD大鼠异动症的发生，这与微球释放的左旋�Objective To investigate the effects of levodopa/benserazide-loaded poly-lactide-coglyeolide （PLGA） mierospheres on motor deficits and levodopa-induced dyskinesia in a rat model of Parkinson＇ s disease （PD） and to explore the mechanisms underlying this effects. Methods The content of levodopa/benserazide released from the microspheres was determined by high-performance liquid chromatography. The rat model of PD was induced by 6-OHDA injections. Then the valid PD rats were treated with levodopa （ 12 mg/kg, s.c. ）/benserazide （ 15 mg/kg, s.c. ） or microspheres. Forepaw adjusting steps were measured on 1, 4, 7, 10 and 14 days after treatment. After 2 weeks of treatment, the abnormal involuntary movements （AIM） were measured. Phosphorylated dopamine, cAMP-regulated phosphoprotein of 32 kDa （DARPP-32） at threonine 34 levels were determined by immunohistochemistry and Western blot respectively. In addition, the levels of AFosB were measured by Western blot. Results In vivo release test showed that 76. 2% of levodopa and 83.6% benserazide were released from the microspheres on day 7. Forepaw adjusting steps showed that the scores of forepaw adjusting steps in microspheres-treated PD rats were （5.8 ± 1.6 ） and （5.2 ± 1.5 ） respectively on 10 and 14 days of treatment, which were increased compared to levodopa-treaded PD rats （2. 4 ± 1.1 and 1.2 ± 0. 5; t = 4. 12, 5.43, all P 〈 0. 01 ）. The AIM scores of microspheres-treated rats （ 16.0 ± 2. 1 ） were decreased significantly compared to levodopa-treated rats （26. 0 ± 3.2） on day 14 （t = 6. 59, P 〈 0. 01 ）. Immunohistochemistry indicated that the phosphorylated levels of DARPP-32 in microspheres-treated rats （ （ 3.7 ± 1.3 ） ×10^4 ） were decreased significantly compared to levodopa-treated rats （ （7.9 ± 2. 2） ×10^4 ; t = 2. 95,P 〈 0. 05 ）. In addition, Western blot showed that the levels of phosphorylated DARPP-32 and AFosB were 119.4% ± 11.3% and 149. 3% ± 12. 3% respectively, which were decr

关 键 词：帕金森病 运动失调 左旋多巴 苄丝肼 微球体 

分 类 号：R742.5[医药卫生—神经病学与精神病学]