中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (4): 854-866.doi: 10.4103/1673-5374.321001

• 原著:退行性病与再生 • 上一篇    下一篇

神经降压素-多聚体纳米颗粒介导人脑多巴胺神经营养因子基因逆转黑质纹状体的神经变性

  

  • 出版日期:2022-04-15 发布日期:2021-10-18

Cerebral dopamine neurotrophic factor transfection in dopamine neurons using neurotensin-polyplex nanoparticles reverses 6-hydroxydopamine-induced nigrostriatal neurodegeneration

Manuel A. Fernandez-Parrilla1, David Reyes-Corona1, Yazmin M. Flores-Martinez2, Rasajna Nadella3, Michael J. Bannon4, Lourdes Escobedo1, Minerva Maldonado-Berny1, Jaime Santoyo-Salazar5, Luis O. Soto-Rojas6, Claudia Luna-Herrera7, Jose Ayala-Davila1, Juan A. Gonzalez-Barrios8, Gonzalo Flores9, Maria E. Gutierrez-Castillo10, Armando J. Espadas-Alvarez10, Irma A. Martínez-Dávila1, Porfirio Nava1, Daniel Martinez-Fong1, 11, *   

  1. 1Departamento de Fisiología, Biofísica y Neurociencias, Centro de Investigación y de Estudios Avanzados, Ciudad de México, México; 2Programa Institucional de Biomedicina Molecular, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Ciudad de México, México; 3Department of Biosciences, IIIT-Srikakulam, Rajiv Gandhi University of Knowledge Technologies (RGUKT), Andhra Pradesh, India; 4Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI, USA; 5Departamento de Física, Centro de Investigación y de Estudios Avanzados, Ciudad de México, México; 6Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla de Baz, Edo. de México, México; 7Departamento de Fisiología, Escuela Nacional de Ciencias Biológicas, Ciudad de México, México; 8Laboratorio de Medicina Genómica, Hospital Regional “1° de Octubre”, ISSSTE, Ciudad de México, México; 9Laboratorio de Neuropsiquiatría, Instituto de Fisiología, Benemérita Universidad Autónoma de Puebla, Puebla, Puebla, México; 10Departamento de Biociencias e Ingeniería, Centro Interdisciplinario de Investigaciones y Estudios sobre Medio Ambiente y Desarrollo, Instituto Politécnico Nacional, Ciudad de México, México; 11Programa de Nanociencias y nanotecnología, Centro de Investigación y de Estudios Avanzados, Ciudad de México, México
  • Online:2022-04-15 Published:2021-10-18
  • Contact: Daniel Martinez-Fong, MD, PhD, martinez.fong@gmail.com.
  • Supported by:
    This study was supported by the Consejo Nacional de Ciencia Tecnología (Conacyt) de México (Grant # 254686, to DMF). 

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摘要：

有研究指出，黑质多巴胺神经元中神经营养因子的过表达是逆转帕金森病黑质纹状体多巴胺系统神经变性的有前途的方法。实验旨在了解神经降压素-多聚体纳米颗粒系统介导人脑多巴胺神经营养因子的表达是否可以逆转6-羟基多巴胺诱导的多巴胺神经元损伤。（1）3种电子显微镜技术显示，表达人脑多巴胺神经营养因子基因的神经降压素-多聚体纳米颗粒尺寸为20-150 nm，并且此类纳米颗粒能够在体外表达可分泌的人脑多巴胺神经营养因子；（2）动物实验进一步在大鼠6-羟基多巴胺诱导黑质纹状体系统损伤后第21天将该纳米颗粒注射入黑质致密部，发现多巴胺神经元中可检测到人脑多巴胺神经营养因子，在整个实验过程中黑质致密部和纹状体中其水平保持恒定；（3）人脑多巴胺神经营养因子转染后黑质致密部和纹状体中酪氨酸羟化酶阳性（TH+）黑质细胞群和TH+纤维密度增加，并伴随着βIII微管蛋白和与生长相关的蛋白43磷酸化S41（GAP43p），以及多巴胺及其分解代谢产物水平的增加，苯丙胺诱导的大鼠旋转行为，以及自发运动和非运动行为明显改善；（4）说明神经降压素-多聚体纳米颗粒系统介导人脑多巴胺神经营养因子对帕金森病黑质纹状体多巴胺系统起到了神经营养作用，纳米颗粒介导的人脑多巴胺神经营养因子基因递送有潜力用于帕金森病的治疗。

https://orcid.org/0000-0002-2934-8380 (Daniel Martinez-Fong)

Abstract: Overexpression of neurotrophic factors in nigral dopamine neurons is a promising approach to reverse neurodegeneration of the nigrostriatal dopamine system, a hallmark in Parkinson’s disease. The human cerebral dopamine neurotrophic factor (hCDNF) has recently emerged as a strong candidate for Parkinson’s disease therapy. This study shows that hCDNF expression in dopamine neurons using the neurotensin-polyplex nanoparticle system reverses 6-hydroxydopamine-induced morphological, biochemical, and behavioral alterations. Three independent electron microscopy techniques showed that the neurotensin-polyplex nanoparticles containing the hCDNF gene, ranging in size from 20 to 150 nm, enabled the expression of a secretable hCDNF in vitro. Their injection in the substantia nigra compacta on day 21 after the 6-hydroxydopamine lesion resulted in detectable hCDNF in dopamine neurons, whose levels remained constant throughout the study in the substantia nigra compacta and striatum. Compared with the lesioned group, tyrosine hydroxylase-positive (TH+) nigral cell population and TH+ fiber density rose in the substantia nigra compacta and striatum after hCDNF transfection. An increase in βIII-tubulin and growth-associated protein 43 phospho-S41 (GAP43p) followed TH+ cell recovery, as well as dopamine and its catabolite levels. Partial reversal (80%) of drug-activated circling behavior and full recovery of spontaneous motor and non-motor behavior were achieved. Brain-derived neurotrophic factor recovery in dopamine neurons that also occurred suggests its participation in the neurotrophic effects. These findings support the potential of nanoparticle-mediated hCDNF gene delivery to develop a disease-modifying treatment against Parkinson’s disease. The Institutional Animal Care and Use Committee of Centro de Investigación y de Estudios Avanzados approved our experimental procedures for animal use (authorization No. 162-15) on June 9, 2019.

Key words: axonal growth, brain-derived neurotrophic factor, gene delivery, nanoparticles, neuritogenesis, neuronal cytoskeleton, neuroregeneration, neurorestoration, neurotrophic therapy, Parkinson’s disease, reinnervation, substantia nigra

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