Neuroprotective and neuroregenerative effects of trophic factors

营养因子的神经保护和神经再生作用

• 批准号: 8553232
• 负责人: Yun Wang
• 金额: $ 54.65万
• 依托单位: NATIONAL INSTITUTE ON DRUG ABUSE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8553232
• 关键词: Adolescence; Adolescent; Adult; Adverse effects; Aftercare; Animals; Astrocytes; Attenuated; Autopsy; Behavior; Behavioral; Bilateral; Brain; Brain Injuries; Brain Ischemia; Cell Proliferation; Cells; Cerebral Infarction; Cerebral cortex; Cerebrum; Complementary DNA; Corpus striatum structure; DNA Fragmentation; Data; Development; Dopamine; Dose; Fiber; Gene Delivery; Genes; Genetic Polymorphism; Genetic Predisposition to Disease; Human; In Situ Nick-End Labeling; Individual; Infarction; Inflammation; Injection of therapeutic agent; Injury; Intoxication; Ischemic Brain Injury; Label; Lead; Lesion; Life; Long-Term Effects; Maintenance; Measures; Mediating; Messenger RNA; Metabolism; Methamphetamine; Midbrain structure; Modeling; Motor; Motor Activity; Mus; Mutation; Nerve Regeneration; Neuroglia; Neurologic; Neuronal Injury; Neurons; Neurotoxins; Oxidopamine; Parkinson Disease; Phenotype; Predisposition; Property; Proteins; Rattus; Recovery; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Stress; Stroke; Substantia nigra structure; Symptoms; System; Time; Toxic effect; Toxin; Tyrosine 3-Monooxygenase; adeno-associated viral vector; alitretinoin; alpha synuclein; base; bone morphogenetic protein 7; brain tissue; central nervous system injury; density; dopamine system; dopamine transporter; dopaminergic neuron; endoplasmic reticulum stress; functional outcomes; immunoreactivity; improved; in vivo; mRNA Expression; methamphetamine exposure; motor deficit; nervous system development; neuronal survival; neurorestoration; neurotrophic factor; nigrostriatal pathway; pars compacta; prevent; protective effect; receptor; regenerative; response; treatment strategy

1. BMP7: In primary dopaminergic neuronal culture, BMP7 reduced MA mediated toxicity (i.e. decreased TH immunoreactivity and increasing TUNEL labeling). BMP7 also reduced MA mediated toxicity in vivo. Intra-cerebroventricular administration of BMP7 antagonized MA-induced changes in TH immunoreactivity in striatum and locomotor activity in mice. We found that intracerebroventricular administration of 9-cis-retinoic acid (9cRA) enhanced BMP7 mRNA expression, detected by RTPCR. Pretreatment with 9cRA attenuated the loss of TH immunoreactivity in striatum after high dose of MA administration. In stroke rats, pretreatment with 9cRA increased locomotor activity and attenuated neurological deficits 2 days after stroke. 9cRA also reduced cerebral infarction and TUNEL labeling. These protective responses were antagonized by BMP antagonist noggin given at one day after 9cRA injection. Taken together, our data suggest that 9cRA has protective effects and these effects involve BMPs. 2. MANF: We first examined the interaction of MANF and methamphetamine (MA) in primary ventral mesencephalic cultures containing dopaminergic neurons. Our preliminary data indicate that MANF attenuated MA toxicity in dopaminergic neurons in culture. We also found that MANF reduced ischemic brain injury and promoted behavioral recovery. Pre-stroke delivery of MANF protein, at a dose of 6 g, reduced cerebral infarction, suppressed DNA fragmentation, and facilitated motor recovery in stroke rats. Local administration of AAV-MANF enhanced MANF expression in neurons and glia in cerebral cortex. Pretreatment with AAV-MANF reduced the volume of cerebral infarction and facilitated behavioral recovery in stroke rats. Our data suggest that administration of either MANF protein or AAV-MANF reduces ischemic brain injury. 3. CDNF: We examined whether CDNF injections into striatum of C57/Bl6 mice have neuroprotective and neurorestorative properties for the nigrostriatal dopamine system after MPTP injections. We found that bilateral striatal CDNF injections, given 20-h before MPTP exposure, improved horizontal and vertical motor behavior when measured 2 weeks afterwards. In addition, CDNF pre-treatment increased tyrosine hydroxylase (TH)-immunoreactivity in the striatum and in the substantia nigra pars reticulata (SNpr), as well as number of TH-positive cells in substantia nigra pars compacta (SNpc). Post-treatment with CDNF, given 1 week after MPTP injections, increased horizontal and vertical behavior of mice. Furthermore, dopamine fiber densities in striatum and the number of TH positive cells in SNpc were increased after CDNF injections. We conclude that intrastriatal CDNF administration is both neuroprotective and neurorestorative for the nigrostriatal dopamine system in the MPTP model, which supports the development of CDNF-based treatment strategies for Parkinsons disease. 4. Nurr1: We found that METH binge exposure in adolescence led to greater damage in the nigrostrial dopaminergic system when mice were exposed to METH binge later in life, suggesting a long-term adverse effect on the dopaminergic system. Compared to naive mice that received METH binge treatment for the first time, mice pretreated with METH in adolescence showed a greater loss of tyrosine hydroxylase (TH) immunoreactivity in striatum, loss of THir fibers in the substantia nigra reticulata (SNr) as well as decreased dopamine transporter (DAT) level and compromised DA clearance in striatum. These effects were further exacerbated in Nurr1 heterozygous mice. Our data suggest that a prolonged adverse effect exists following adolescent METH binge exposure which may lead to greater damage to the dopaminergic system when exposed to repeated METH later in life. Furthermore, our data support that Nurr1 mutations or deficiency could be a potential genetic predisposition which may lead to higher vulnerability in some individuals.

1. BMP 7：在原代多巴胺能神经元培养物中，BMP 7降低MA介导的毒性（即降低TH免疫反应性和增加TUNEL标记）。BMP 7还降低了MA介导的体内毒性。脑室注射BMP 7可拮抗MA诱导的小鼠纹状体TH免疫反应性和自发活动的变化。我们发现，侧脑室注射9-顺式维甲酸（9 cRA）增强BMP 7 mRNA的表达，检测到的RTPCR。9 cRA预处理可减轻大剂量MA后纹状体TH免疫反应性的丧失。在中风大鼠中，用9 cRA预处理增加中风后2天的自发活动和减弱神经功能缺损。9 cRA还可减少脑梗死和TUNEL标记。在注射9 cRA后1天给予BMP拮抗剂noggin可拮抗这些保护作用。总之，我们的数据表明，9 cRA具有保护作用，这些作用涉及BMP。 2. MANF：我们首先研究了MANF和甲基苯丙胺（MA）在含有多巴胺能神经元的初级腹侧中脑培养物中的相互作用。 我们的初步数据表明，MANF衰减MA毒性多巴胺能神经元的文化。 我们还发现，MANF减少缺血性脑损伤，促进行为恢复。 在中风大鼠中，中风前给予MANF蛋白，剂量为6 g，减少脑梗死，抑制DNA片段化，并促进运动恢复。局部应用AAV-MANF可增强大脑皮层神经元和胶质细胞中MANF的表达。AAV-MANF预处理可减少脑梗死体积，促进脑梗死大鼠行为学恢复。我们的数据表明，MANF蛋白或AAV-MANF的管理减少缺血性脑损伤。 3. CDNF：我们研究了C57/B16小鼠纹状体注射CDNF是否对MPTP注射后的黑质纹状体多巴胺系统具有神经保护和神经恢复特性。我们发现，双侧纹状体CDNF注射，给予MPTP曝光前20小时，改善水平和垂直运动行为时，测量2周后。此外，CDNF预处理增加了纹状体和黑质网状部（SNpr）中的酪氨酸羟化酶（TH）免疫反应性，以及黑质网状部（SNpc）中TH阳性细胞的数量。在MPTP注射后1周给予CDNF后处理，增加了小鼠的水平和垂直行为。此外，注射CDNF后，纹状体多巴胺纤维密度和SNpc中TH阳性细胞数量增加。我们的结论是，纹状体内CDNF管理是神经保护和神经恢复的黑质纹状体多巴胺系统的MPTP模型，这支持了帕金森病的CDNF为基础的治疗策略的发展。 4.护士1：我们发现，当小鼠在生命后期暴露于METH狂欢时，青春期的METH狂欢暴露会导致黑质多巴胺能系统的更大损伤，这表明对多巴胺能系统的长期不良影响。与首次接受METH狂欢治疗的幼稚小鼠相比，在青春期用METH预处理的小鼠显示出纹状体中酪氨酸羟化酶（TH）免疫反应性的更大损失、黑质网状（SNr）中THir纤维的损失以及纹状体中多巴胺转运蛋白（DAT）水平的降低和DA清除率的受损。这些影响在Nurr 1杂合子小鼠中进一步加剧。我们的数据表明，一个长期的不良反应存在以下青少年甲基狂欢暴露，这可能会导致更大的损害多巴胺能系统时，暴露于重复甲基以后的生活。此外，我们的数据支持Nurr 1突变或缺陷可能是一种潜在的遗传易感性，可能导致某些个体更高的脆弱性。