Neuroprotective and regenerative effects of small molecules and their receptors

小分子及其受体的神经保护和再生作用

• 批准号: 8336439
• 负责人: Yun Wang
• 金额: $ 43.32万
• 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/8336439
• 关键词: 3,4-Dihydroxyphenylacetic Acid; 3-nitropropionate; Ablation; Adenosine; Adenosine A3 Receptor; Adult; Aftercare; Agonist; Amphetamines; Animals; Anisotropy; Anoxia; Apoptosis; Apoptotic; Attenuated; Behavioral; Binding; Biochemical; Blood - brain barrier anatomy; Brain; Brain Injuries; Brain-Derived Neurotrophic Factor; Calcium Binding; Carbon Dioxide; Cell Death; Cell Nucleus; Cell Survival; Cells; Cerebral Infarction; Cerebrum; Cleaved cell; Contralateral; Corpus striatum structure; Cultured Cells; DNA; Data; Development; Diffusion Magnetic Resonance Imaging; Dopamine; Dose; Embolism; Exposure to; FOS gene; Fiber; Follow-Up Studies; Glucose; Growth; Heart; High Pressure Liquid Chromatography; Hippocampus (Brain); Hour; Hydrogen Peroxide; Hypoxia; Image; Immune response; Immune system; In Situ Nick-End Labeling; Infarction; Inflammation; Inflammatory; Injection of therapeutic agent; Injury; Intoxication; Ipsilateral; Ischemia; Ischemic Brain Injury; Isomerism; JUN gene; Label; Lesion; Lung; MAPK8 gene; Matrix Metalloproteinases; Measures; Mediating; Mediator of activation protein; Methamphetamine; Microglia; Middle Cerebral Artery Occlusion; Mitochondria; Motor Activity; Mus; Mutant Strains Mice; Naloxone; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurologic; Neuronal Dysfunction; Neurons; Neurotoxins; PC12 Cells; Parkinsonian Disorders; Pharmaceutical Preparations; Play; Process; Rattus; Recovery; Reporting; Reserpine; Rodent Model; Role; Saline; Side; Signal Transduction; Site; Slice; Stroke; Synaptophysin; TNF gene; Tissues; Toxic effect; Transplantation; Tretinoin; Up-Regulation; Vitamin A; Weight; alitretinoin; body asymmetry; bone morphogenetic protein 7; brain tissue; caspase-3; density; deprivation; dopaminergic neuron; immunoreactivity; improved; in vivo; inflammatory marker; inhibitor/antagonist; mRNA Expression; male; mesangial cell; middle cerebral artery; nervous system disorder; neurofilament; novel therapeutic intervention; pifithrin; post stroke; preconditioning; prevent; protective effect; receptor; regenerative; reinnervation; relating to nervous system; repaired; research study; response; small molecule; synaptogenesis

(1) Amphetamine in neuroregeneration: AM post-treatment significantly reduced neurological deficits, as measured by body asymmetry and Bedersons score in stroke rats. T2WI and diffusion tensor imaging (DTI) were used to examine the size of infarction and axonal reinnervation, respectively, before and following treatment on days 2, 10 and 25 after stroke. AM treatment reduced the volume of tissue loss on days 10 and 25. A significant increase in fractional anisotropy ratio was found in the ipislateral cortex after repeated AM administration, suggesting a possible increase in axonal outgrowth in the lesioned side cortex. Western analysis indicated that AM significantly increased the expression of synaptophysin ipsilaterally and neurofilament bilaterally. AM also enhanced matrix metalloproteinase (MMP) enzymatic activity, determined by MMP zymography in the lesioned side cortex. qRT-PCR was used to examine the expression of trophic factors after the 1st and 2nd doses of AM or saline injection. The expression of BDNF, but not BMP7 or CART, was significantly enhanced by AM in the lesioned side cortex. In conclusion, post-stroke treatment with AM facilitates behavioral recovery, which is associated with an increase in fractional anisotropy activity, enhanced fiber growth in tractography, synaptogenesis, upregulation of BDNF, and MMP activity mainly in the lesioned cortex. Our data suggest that the ipsilateral cortex may be the major target of action in stroke brain after AM treatment. (2) adenosine A3R receptor: We reported that high doses of Meth suppressed locomotor activity in adult male A3R null mutant (-/-) mice and their controls (+/+), with a greater reduction being found in the -/- mice. Brain tissues were collected at 3 days after the Meth or saline injections. Meth treatment reduced striatal dopamine (DA) levels in both +/+ and -/- mice, examined by HPLC, with an increase in DOPAC/DA ratio being found only in -/- animals after Meth treatment. Meth also significantly increased ionized calcium-binding adaptor molecule 1 (Iba-1) and cleaved caspase-3 level in striatum as well as Iba-1 and TNF mRNA expression in nigra in -/-, compared to +/+, mice. Previous studies have shown that pharmacological suppression of VMAT2 by reserpine enhanced Meth toxicity by increasing cytosolic DA and inflammation. A significant reduction in striatal VMAT2 expression was found in -/- mice, compared to +/+ mice, suggesting that increase in sensitivity to Meth injury in -/- mice may be related to a reduction in VMAT2 expression in these mice. Our data suggest that A3R -/- mice are more sensitive to high doses of Meth. (3) pifithrin-a (PFT-a): (A) We found that PFTa has protective against Meth -mediated injury in dopaminergic neurons. High dose of Meth reduced TH immunoreactivity and fiber density in primary dopaminergic neuronal culture. Co-treatment with PFTa significantly attenuated these degenerative changes. Our data suggest that treatment with a p53 inhibitor or suppression of p53 expression limits neuronal dysfunction and cell death induced by exposure to dopaminergic neurotoxins. (B) Systemic administration of PFT- enhanced the survival of dopaminergic neuronal transplants in vivo. PFT- treatment suppressed cell death, increased survival of TH (+) cells, enhanced TH neurite outgrowth from ventromesencephalic grafts, and augmented behavioral recovery in Parkinsonian rats. In conclusion, our data suggest that activation of p53 is an important mediator for cell death after injury or during neural repair. Transient suppression of p53 may increase the survival of neurons or NPCs after brain injury. (4) Retinoic acid (RA): Pretreatment with 9 cis RA (9cRA) increased locomotor activity, attenuated neurological deficits, and reduced cerebral infarction and TUNEL labeling in stroke rats. 9cRA increased the expression of bone morphogenetic protein 7 (BMP7) in brain. The protective response of 9cRA can be antagonized by BMP antagonist noggin. Our data suggest that RA can induce protective responses against Meth toxicity in cultured cells and in vivo, possibly through BMP7 mechanism. (5) (-)Naloxone: Neurodegeneration can occur through the activation of inflammatory processes. From experiments conducted this past year, we have observed improved behavioral recovery (body asymmetery and neurological score) following stroke when (+)naloxone was administered daily for 1 week starting one day after middle cerebral artery occlusion in rats. We are currently following up these studies comparing (-)naloxone as well as examining changes in markers of inflammation that are upregulated following ischemia of the stroke.

（1）神经变成中的苯丙胺：AM治疗显着降低了神经功能缺陷，如人体不对称和中风大鼠的贝德森评分所测量的那样。 T2WI和扩散张量成像（DTI）分别检查梗塞的大小和轴突加剧的大小，并在中风后第2、10和25天进行治疗前后。 AM治疗减少了第10天和第25天的组织损失的体积。在重复AM给药后，在iPislateral皮层中发现了分数各向异性比例的显着增加，这表明在病变的侧面皮层中可能增加轴突产物的增长。西方的分析表明，双侧和神经丝的表达显着增加了突触素的表达。 AM还增强了通过MMP酶学在病变的侧皮层中确定的基质金属蛋白酶（MMP）酶活性。 QRT-PCR用于检查AM或盐水注射剂或第二剂量后的营养因子的表达。在病变的侧皮层中，AM显着增强了BDNF而不是BMP7或CART的表达。总之，卒中后治疗的AM促进了行为恢复，这与分数各向异性活性的增加，拖拉术，突触发生，BDNF上调和MMP活性的增加有关，主要是病变的皮质中。 我们的数据表明，同侧皮层可能是AM治疗后中风大脑中的主要作用目标。 （2）腺苷A3R受体：我们报道了成年雄性A3R无效突变体（ - / - ）小鼠及其对照（+/+）的高剂量的MET抑制运动活性，并且在 - / - 小鼠中发现了更大的降低。在甲基甲基或盐水注射后3天收集脑组织。 通过HPLC检查的 +/ +和 - / - 小鼠的曲纹多巴胺（DA）水平降低了纹状体多巴胺（DA）水平，仅在甲基甲基治疗后仅在 - / - 动物中发现dopac/da比的增加。与 +/ +小鼠相比，METH还显着增加了纹状体中的离子钙结合衔接子分子1（IBA-1）和纹状体中的caspase-3水平以及nigra中的IBA-1和TNF mRNA表达，与 +/ +小鼠相比。先前的研究表明，通过增加胞质DA和炎症，RESERPINE对RESERPINE对VMAT2的药理抑制促进了METH毒性。与 +/ +小鼠相比，在 - / - 小鼠中发现纹状体VMAT2表达的显着降低，这表明 - / - 小鼠对甲基损伤的敏感性增加可能与这些小鼠中VMAT2表达的降低有关。 我们的数据表明，A3R - / - 小鼠对高剂量的甲基苯酚更敏感。 （3）pifithrin-a（pft-a）：（a）我们发现PFTA对多巴胺能神经元中的甲基介导的损伤具有保护作用。高剂量的METH降低了原发性多巴胺能神经元培养的TH免疫反应性和纤维密度。与PFTA共同治疗显着减弱了这些退化性变化。我们的数据表明，用p53抑制剂或p53表达抑制的治疗限制了暴露于多巴胺能神经毒素引起的神经元功能障碍和细胞死亡。 （b）PFT的系统给药增强了体内多巴胺能神经元移植的存活。 PFT-治疗抑制了细胞死亡，Th（+）细胞的存活增加，腹脑移植的TH神经突生长增长以及帕金森氏大鼠的行为恢复增强。总之，我们的数据表明，p53的激活是受伤后或神经修复期间细胞死亡的重要介体。 p53的短暂抑制可能会增加脑损伤后神经元或NPC的存活。 （4）视黄酸（RA）：带有9个顺式RA（9cra）的预处理增加了运动活性，神经系统缺陷减弱，脑梗死减少和中风大鼠的TUNEL标记。 9cra增加了脑中骨形态发生蛋白7（BMP7）的表达。 BMP拮抗剂Noggin可以拮抗9cra的保护反应。我们的数据表明，RA可以通过BMP7机制诱导针对培养细胞和体内MET的毒性的保护反应。 （5）（ - ）纳洛酮：神经变性可以通过激活炎症过程而发生。从过去一年进行的实验中，我们观察到中风后的行为恢复改善（身体不对称和神经学评分），当时（+）纳洛酮每天在大鼠中大脑中动脉闭塞后一天开始（+）纳洛酮。 我们目前正在跟踪这些研究，以比较（ - ）纳洛酮以及检查炎症标志物的变化，这些变化在中风的缺血之后被上调。