侧脑室注射腺病毒介导的GDNF基因对帕金森病的保护作用

doi:10.6040/j.issn.1671-7554.0.2015.1321

摘要/Abstract

摘要： 目的探讨腺病毒介导的神经胶质细胞源性神经营养因子(GDNF)基因脑内转移对帕金森病(PD)的保护作用。方法采用C57Bl/6小鼠1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-henyl-l,2,3,6-tetrahydropyridine, MPTP)法建立PD模型,随机分为重组GDNF腺病毒(Ad-GDNF)实验组和对照组。Ad-GDNF实验组将Ad-GDNF定向注射至侧脑室,对照组将无GDNF基因的腺病毒定向注射至侧脑室,并测量体质量,进行行为学实验评分,行中脑黑质酪氨酸羟化酶(TH)免疫组化染色,高压液相色谱-电化学仪(HPLC-ECD)检测纹状体多巴胺(DA)、5-羟色胺(5-HT)及5-羟吲哚乙酸(5-HIAA)含量,采用ELISA、RT-PCR法检测Ad-GDNF在中脑的表达。结果侧脑室注射病毒后2周,Ad-GDNF实验组体质量、行为学实验得分、黑质酪氨酸羟化酶(TH)阳性细胞、纹状体DA含量均显著高于对照组(P<0.05);Ad-GDNF实验组在中脑内有过表达,中脑GDNF含量约是对照组2倍。结论侧脑室注射腺病毒介导的GDNF基因可减轻1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-henyl-l,2,3,6-tetrahydropyridine, MPTP)诱发的小鼠DA能神经元进行性变性,保护DA能神经元,提示这一手段在PD保护性治疗方面具有一定的应用价值。

关键词: 6-四氢吡啶, 神经胶质细胞源性神经营养因子, 腺病毒, 帕金森病, 基因治疗, 3, 1-甲基-4-苯基-1, 2

Abstract: Objective To investigate the neuroprotective effect of adenovirus-mediated glial cell line-derived neurotrophic factor(GDNF)gene intracerebrally transferred on the treatment of Parkinsons disease(PD). Methods PD models were established by intraperitoneal injection of 1-methyl-4-henyl-l,2,3,6-tetrahydropyridine(MPTP)［30 mg/(kg·d)in C57Bl/6 mice. The models were then divided into Ad-GDNF experimental group and control group. Ad-GDNF was injected into the lateral ventricle of mice in the experimental group. Negative virus was injected into the lateral ventricle of mice in the control group. The body weight of all mice was measured. The neuroprotective effect of Ad-GDNF was evaluated with behavioral tests, immune histochemical assay of the tyrosine hydroxylase(TH)-positive neurons in the substance nigra and measurement of monoamine level in the striatum. The expression of the exogenous GDNF gene in the brain was detected with ELISA and RT-PCR. Results Compared with control group, the experimental group showed improved motor functions(P<0.01)and less loss of body weight(P<0.05), better survival of TH-positive cells(P<0.05), higher DA levels(P<0.05)after two weeks of intracerebroventricular injection. The exogenous 山东大学学报 (医学版)54卷4期 -王思,等.侧脑室注射腺病毒介导的GDNF基因对帕金森病的保护作用 \=-GDNF gene was efficiently over expressed in the experimental group. GDNF protein level in midbrain of the experimental group was twice of the control group. Conclusion Intracerebroventricular injection of adenovirus mediated GDNF gene significantly can protect the dopaminergic neurons of nigrostriatal system from MPTP-induced injury and is conducive in the treatment of Parkinsons disease.

Key words: Glial cell line-derived neurotrophic factor, Adenovirus, Parkinsons disease, Gene therapy, 1-methyl-4-henyl-l,2,3,6-tetrahydropyridine

中图分类号:

R741.02

王思,李秀华,杜鹃,岳龙涛,王瑶,刘菲,郭配. 侧脑室注射腺病毒介导的GDNF基因对帕金森病的保护作用[J]. 山东大学学报（医学版）, 2016, 54(4): 32-36.

WANG Si, LI Xiuhua, DU Juan, YUE Longtao, WANG Yao, LIU Fei, GUO Pei. Neuroprotective effect of adenovirus-mediated GDNF gene in mice models of Parkinsons disease[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(4): 32-36.

参考文献

[1] Stacy M. Medical treatment of Parkinson disease[J]. Neurol Clin, 2009, 27(3): 605-631.
[2] Lin LF, Doherty DH, Lile JD, et al. GDNF: a glial cell line derived neurotrophic factor for midbrain dopaminergic neurons[J]. Science, 1993, 260(5111): 1130-1132.
[3] Lu-Nguyen NB, Broadstock M, Schliesser MG, et al. Transgenic expression of human glial cell line-derived neurotrophic factor from integration-deficient lentiviral vectors is neuroprotective in a rodent model of Parkinsons disease[J]. Hum Gene Ther, 2014, 25(7): 631-641.
[4] Gash DM, Zhang Z, Ovadia A, et al. Functional recovery in parkinsonian monkeys treated with GDNF[J]. Nature, 1996, 380(6571): 252-255.
[5] Cohen AD, Zigmond MJ, Smith AD, et al. Effects of intrastriatal GDNF on the response of dopamine neurons to 6-hydroxydopamine: Time course of protection and neurorestoration[J]. Brain Res, 2011, 1370(2011): 80-88.
[6] 刘洪梅, 丁艳霞, 王炎强, 等. 表皮生长因子加强胶质细胞系源性神经营养因子对PD 模型大鼠黑质多巴胺能神经元的保护作用[J]. 解剖学杂志, 2011, 34(5): 537-575. LIU Hongmei, DING Yanxia, WANG Yanqiang, et al. EGF enhances the effect of GDNF on protecting dopamine neurons in substantia nigra of Parkinsons disease model in rats[J]. Chinese Journal of Anatomy, 2011, 34(5): 537-575.
[7] Gonzalez-Aparicio R, Flores JA, Fernandez-Espejo E. Antiparkinsonian trophic action of glial cell line-derived neurotrophic factor and transforming growth factor β1 is enhanced after co-infusion in rats[J]. ExpNeurol, 2010, 226(1): 136-147.
[8] Dunnett SB, Carter RJ, Watts C, et al. Striatal transplantation in a transgenic mouse model of Huntington's disease[J]. ExpNeurol, 1998, 154(1): 31-40.
[9] Ogawa N, Hirose Y, Ohara S, et al. A simple quantitative bradykinesia test in MPTP-treated mice[J]. Res Commun Chem Pathol Pharmacol, 1985, 50(3): 435-441.
[10] Olsson M, Nikkhah G, Bentlage C, et al. Forelimb akinesia in the rat Parkinson model: differential effects of dopamine agonists and nigral transplants as assessed by a new stepping test[J]. J Neurosci, 1995, 15(2): 3863-3875.
[11] Wesson DW, Wilson DA. Age and gene overexpression interact to abolish nesting behavior in Tg2576 amyloid precursor protein(APP)mice[J]. Behav Brain Res, 2011, 216(1): 408-413.
[12] Tomac A, Lindqvist E, Lin LF, et al.Protection and repair of the nigrostriatal dopaminergic system by GDNF in vivo[J]. Nature, 1995, 373(6512): 335-339.
[13] Bernheimer H, Birkmayer W, Hornykiewicz O, et al. Brain dopamine and the syndromes of Parkinson and Huntington. Clinical, morphological and neurochemical correlations[J]. J Neurol Sci, 1973, 20(4): 415-455.
[14] Littrell OM, Granholm AC, Gerhardt GA, et al. Glial cell-line derived neurotrophic factor(GDNF)replacement attenuates motor impairments and nigrostriatal dopamine deficits in 12-month-old mice with a partial deletion of GDNF[J]. Pharmacol Biochem Behav, 2013, 104(3): 10-19.
[15] Tereshchenko J, Maddalena A, Bähr M, et al. Pharmacologically controlled, discontinuous GDNF gene therapy restores motor function in a rat model of Parkinsons disease[J]. Neurobiol Dis, 2014, 65(5): 35-42.
[16] Cash R, Raisman R, Ploska A, et al. High and low affinity[³H] imipramine binding sites in control and parkinsonian brains[J]. Eur J Pharmacol, 1985, 117(1): 71-80.
[17] Lelieveld IM, Müller ML, Bohnen NI, et al. The role of serotonin in sleep disordered breathing associated with Parkinson disease: a correlative[¹¹C] DASB PET imaging study[J]. PLoS One, 2012, 7(7): e40166.
[18] Biju K, Zhou Q, Li G, et al. Macrophage-mediated GDNF delivery protects against dopaminergic neurodegeneration: a therapeutic strategy for Parkinsons disease[J]. Molecular Therapy, 2010, 18(8): 1536-1544.
[19] Pardo J, Morel GR, Astiz M, et al. Gene Therapy and Cell Reprogramming for the Aging Brain: Achievements and Promise[J]. Curr Gene Ther, 2014, 14(1): 24-34.
[20] Lu-Nguyen NB, Broadstock M, Schliesser MG, et al. Transgenic expression of human glial cell line-derived neurotrophic factor from integration-deficient lentiviral vectors is neuroprotective in a rodent model of Parkinson's disease[J]. Hum Gene Ther, 2014, 25(7): 631-641.

相关文章 15

[1]	吴逸南葛志明李方贺红姜虹张运. 自发性高血压大鼠肾脏血管紧张素转换酶2的表达[J]. 山东大学学报(医学版), 2209, 47(6): 5-.
[2]	孙涛张道来谢珊珊王玉卓冯玉新辛华. 酒精对原代培养的神经前体细胞间隙连接蛋白43表达的影响[J]. 山东大学学报(医学版), 2209, 47(6): 20-.
[3]	孙薏丰,高玉,梁永媛,高杨. CPLX2在30例肝癌组织的表达及其对体外细胞增殖与侵袭的影响[J]. 山东大学学报 (医学版), 2020, 1(9): 34-39.
[4]	索东阳,申飞,郭皓,刘力畅,杨惠敏,杨向东. Tim-3在药物性急性肾损伤动物模型中的表达及作用机制[J]. 山东大学学报 (医学版), 2020, 1(7): 1-6.
[5]	张宝文,雷香丽,李瑾娜,罗湘俊,邹容. miR-21-5p靶向调控TIMP3抑制2型糖尿病肾病小鼠肾脏系膜细胞增殖及细胞外基质堆积[J]. 山东大学学报 (医学版), 2020, 1(7): 7-14.
[6]	马青源,蒲沛东,韩飞,王超,朱洲均,王维山,史晨辉. miR-27b-3p调控SMAD1对骨肉瘤细胞增殖、迁移和侵袭作用的影响[J]. 山东大学学报 (医学版), 2020, 1(7): 32-37.
[7]	肖娟,肖强,丛伟,李婷,丁守銮,张媛,邵纯纯,吴梅,刘佳宁,贾红英. 两种甲状腺超声数据报告系统诊断效能的比较[J]. 山东大学学报 (医学版), 2020, 1(7): 53-59.
[8]	徐继禧,陈伟健. 髓内弥漫性中线胶质瘤伴H3 K27M突变1例[J]. 山东大学学报 (医学版), 2020, 1(7): 96-101.
[9]	赵姗姗,李晓梅,梁婷,张超,宋静,侯桂华. 自噬对¹²⁵I-anti-TLR5同种移植排斥靶向显像的影响[J]. 山东大学学报（医学版）, 2017, 55(9): 46-52.
[10]	刘东,朱冬昀,彭长亮,张程,赵杰,高春正. 脊髓损伤修复的复合透明质酸水凝胶支架的构建及其评价[J]. 山东大学学报（医学版）, 2017, 55(9): 53-59.
[11]	黄竹青,吴雪韦,任冬梅. 槲寄生中酚类化学成分的分离鉴定及其对A549细胞的增殖抑制活性[J]. 山东大学学报（医学版）, 2017, 55(8): 35-41.
[12]	姬原原,包景泊,赵旭,曹鲁宁,崔敏,张娜,于灵芝. 白介素17F促进大鼠成骨细胞增殖、矿化和Runx2、Osterix的表达[J]. 山东大学学报（医学版）, 2017, 55(8): 24-29.
[13]	白永娟,庄志超,郝树彬,王莉鸿,李纯,鲁燕,赵丽,王志玉,温红玲. 肠道病毒71型3D重组病毒的拯救[J]. 山东大学学报（医学版）, 2017, 55(8): 7-12.
[14]	勾云, 周波, 魏操,陈运华, 徐利,刘芬, 张春林, 文敏. 硫辛酸对帕金森病大鼠黑质线粒体的保护作用[J]. 山东大学学报（医学版）, 2017, 55(8): 18-23.
[15]	林琦,张颖,袁苑,戴建建,徐瑞彩,杨琦,耿宝成,韩明勇. 肾上腺转移瘤放射性¹²⁵I粒子植入治疗后胃穿孔1例[J]. 山东大学学报（医学版）, 2017, 55(7): 124-129.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed