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背景:帕金森病患者脑内黑质多巴胺能神经元变性导致芳香族氨基酸脱羧酶(aromaticL-aminoaciddecarboxylase,AADC)活性水平的降低及波动是产生帕金森患者临床症状和在L-dopa替代治疗时“症状波动”等不良反应出现的重要原因之一。目的:探讨AADC基因脑内移植对帕金森病的治疗作用及其在左旋多巴治疗过程中的地位,并探讨阳离子脂质体介导的基因治疗方法的有效性。设计:2×2析因设计。地点和材料:中山大学附属第二医院林百欣医学研究中心,材料主要包括人嗜铬细胞瘤组织,TRIzol,真核表达载体质粒pCDNA3,阳离子脂质体,6-OHDA,阿朴吗啡,AADC单克隆抗体,SD大鼠。干预:使用基因克隆、亚克隆技术构建pCDNA3-AADC真核表达载体;将帕金森病模型的SD大鼠随机分为实验组与对照组,分别以pCD-NA3-AADC重组体和pCDNA3空载体进行阳离子脂质体包裹后,注射于帕金森病SD大鼠纹状体内,并在该基础上对两组大鼠分别采用一定浓度的L-dopa进行治疗。采用免疫组化方法确定AADC的表达。主要观察指标:①AADC基因脑内移植后两组大鼠的旋转行为。②采用一定浓度的L-dopa进行治疗对大鼠旋转行为的影响。结果:①阳离子脂质体包裹的pCDNA3-AADC重组体脑内注射后,在3,7,14,21,28d时大鼠的旋转行为获得了一定的改善(P<0.05),分别改善了56.0%,6
BACKGROUND: Degeneration of dopaminergic neurons in the substantia nigra of Parkinson’s disease leads to the reduction and fluctuation of the levels of aromatic amino acid decarboxylase (AADC) activity in patients with Parkinson’s disease and in the treatment of L-dopa replacement therapy Symptoms of fluctuations "and other adverse reactions appear one of the important reasons. Objective: To investigate the therapeutic effect of AADC gene on Parkinson’s disease and its role in the treatment of levodopa, and to explore the effectiveness of cationic liposome-mediated gene therapy. Design: 2 × 2 factorial design. Location and Materials: The Second Affiliated Hospital of Sun Yat-sen, Lin Baxin Medical Research Center, the main materials include human pheochromocytoma tissue, TRIzol, eukaryotic expression vector plasmid pCDNA3, cationic liposomes, 6-OHDA, apomorphine, AADC monoclonal Antibody, SD rat. Intervention: The eukaryotic expression vector pCDNA3-AADC was constructed by gene cloning and subcloning techniques. SD rats were randomly divided into experimental group and control group, and were respectively treated with pCDNA3-AADC recombinant and pCDNA3 empty vector Cationic liposomes were injected into Parkinson’s disease SD rat striatum, and on the basis of the two groups of rats were treated with a certain concentration of L-dopa. Immunohistochemistry was used to determine AADC expression. MAIN OUTCOME MEASURES: ① Rotation behavior of rats in both groups after brain transplantation of AADC gene. ② The effect of treatment with a certain concentration of L-dopa on the rotation behavior of rats. RESULTS: ① After the intraperitoneal injection of cationic liposome-encapsulated pCDNA3-AADC recombinant, the rotational behavior of rats was improved at 3, 7, 14, 21 and 28 days (P <0.05), and the improvement was 56.0 %, 6