【作者】 黄容琴； 蒋晨； 裴元英；

【Author】 Huang Rong-qin, Jiang Chen*, Pei Yuan-ying (Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai 200032, China)

【机构】 复旦大学药学院药剂教研室；

【摘要】 目的以帕金森病(PD)为疾病模型,评价了新型非病毒载体高分子PAMAM-PEG-Lf包载编码人源性胶质细胞源性神经营养因子(GDNF)的治疗基因(hGDNF)的纳米粒在慢性PD模型大鼠上的药效。方法通过酶联免疫吸附法比较不同载体的脑内治疗基因表达以及多次给药的有效性;建立鱼藤酮慢性PD模型,设计多次静脉给药方案,通过行为学评价、免疫组织化学评价和纹状体单胺类神经递质的HPLC测定三个指标,评价Lf修饰的载治疗基因纳米粒在慢性PD模型大鼠上的药效。结果单次给药时,PAMAM-PEG-Lf/hGDNF纳米粒的脑部基因表达最高,且随时间会降低;多次给药可以持续增加纳米粒的脑内基因表达,5次给药的表达量显著高于单次给药或3次给药,证实了多次给药的可行性和有效性。同时,在鱼藤酮慢性PD模型上,多次给予PAMAM-PEG-Lf/hGDNF纳米粒同样显示出优于单次给药的疗效,多次给药能显著改善PD大鼠的行动能力,提高酪氨酸羟化酶(TH)免疫反应性,同时提高纹状体中DA及其代谢产物的水平。结论PAMAM-PEG-Lf/DNA纳米粒是一种高效的非病毒载基因系统,多次静脉注射是一种有效可行的非侵袭性给药方式,可持续提高非病毒载体的脑内基因表达。更多还原

【Abstract】 OBJECTIVE Aim to evaluate the pharmacodynamics of polyamidoamine (PAMAM)-polyethylene glycol (PEG)-lactoferrin (Lf) encapsulating human glial cell line-derived neurotrophic factor gene (hGDNF) in the chronic rat model of Parkinson’s disease. METHODS Enzyme linked-immnuno-sorbent assay was used to compare the brain gene expression of different types of nanoparticles and the feasibility of multiple dosing administration. A multiple dosing regimen was designed for the first time to compare the therapeutic effects of single and multiple dosing administrations in a rotenone-induced chronic PD model. RESULTS The results of single injection showed that PAMAM-PEG-Lf/hGDNF NP had higher brain gene expression compared to other types of nanoparticles, and this gene expression decreased with time. Multiple dosing administrations could enhance and maintain the brain gene products in a higher level. The gene expression of five dosing administrations was significantly higher that that of three dosing administrations. These results demonstrated the feasibility and effectiveness of multiple dosing administrations. In the rotenone-induced chronic PD model, multiple dosing administrations also exhibited better therapeutic effects than single injection. Extending the administration period of PAMAM-PEG-Lf/hGDNF NP could significantly improve locomotor activity, reduce dopaminergic neuronal loss and enhance monoamine neurotransmitter levels on PD rats, which indicating more powerful neuroprotective effects of multiple dosing administrations of Lf-modifined nanoparticles. CONCLUSION All the results clearly demonstrate that PAMAM-PEG-Lf/DNA NP is an efficient non-viral gene delivery system, and multiple dosing intravenous administrations could be used as a practical non-invasive means to enhance and maintain the higher brain gene expression of non-viral gene vectors.更多还原