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.

（一） 安非他明在神经再生中的作用：AM治疗后显着减少神经功能缺损，测量身体不对称性和Bedersons评分中风大鼠。分别于治疗前和治疗后第2、10、25天，采用T2 WI和弥散张量成像（DTI）检查梗死范围和轴索神经再支配情况。AM处理减少了第10天和第25天的组织损失量。一个显着增加分数各向异性比被发现在同侧皮层重复AM管理后，这表明可能增加轴突生长在受损侧皮层。Western分析显示AM显著增加同侧突触素和双侧神经丝的表达。AM还增强了基质金属蛋白酶（MMP）酶活性，通过MMP酶谱测定在受损侧皮质。采用qRT-PCR检测AM或生理盐水注射第1次和第2次给药后营养因子的表达。AM可显著增强损伤侧皮质BDNF的表达，但不增强BMP 7或CART的表达。总之，中风后AM治疗有利于行为恢复，这与分数各向异性活动的增加，纤维束成像，突触发生，BDNF的上调，和MMP活性增强，主要是在受损皮层。 我们的数据表明，同侧皮质可能是中风脑AM治疗后的主要作用靶点。 (2)腺苷A3 R受体： 我们报道了高剂量的甲基抑制成年雄性A3 R无效突变（-/-）小鼠及其对照（+/+）的运动活动，在-/-小鼠中发现了更大的减少。在Meth或盐水注射后3天收集脑组织。 通过HPLC检测，甲基苯丙胺治疗降低了+/+和-/-小鼠的纹状体多巴胺（DA）水平，仅在甲基苯丙胺治疗后的-/-动物中发现DOPAC/DA比率增加。与+/+小鼠相比，甲基也显著增加了纹状体中离子化钙结合接头分子1（Iba-1）和切割的caspase-3水平以及黑质中Iba-1和TNF mRNA的表达。先前的研究表明，利血平对VMAT 2的药理学抑制通过增加胞质DA和炎症增强了Meths毒性。与+/+小鼠相比，在-/-小鼠中发现纹状体VMAT 2表达显著降低，表明-/-小鼠中对甲基损伤的敏感性增加可能与这些小鼠中VMAT 2表达的降低有关。 我们的数据表明，A3 R-/-小鼠对高剂量的甲基更敏感。 （三） 匹非亭-α（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。在原代多巴胺能神经元培养中，高剂量的甲基减少TH免疫反应性和纤维密度。与PFTa联合治疗显著减弱了这些退行性变化。我们的数据表明，用p53抑制剂或抑制p53表达的治疗限制了暴露于多巴胺能神经毒素诱导的神经元功能障碍和细胞死亡。(B)全身给予PFT-1可提高体内多巴胺能神经元移植物的存活率。PFT治疗抑制细胞死亡，增加TH（+）细胞的存活，增强腹中脑移植物的TH神经突生长，并增强帕金森病大鼠的行为恢复。总之，我们的数据表明，激活p53是一个重要的介质损伤后或神经修复过程中的细胞死亡。 p53的瞬时抑制可增加脑损伤后神经元或NPC的存活。 （四） 视黄酸（RA）：9 cis RA（9 cRA）预处理增加自发活动，减轻神经功能缺损，减少脑梗死和TUNEL标记在中风大鼠。9 cRA可增加脑组织骨形态发生蛋白7（BMP 7）的表达。BMP拮抗剂noggin可拮抗9 cRA的保护作用。我们的数据表明，RA可以诱导保护性反应，对甲基毒性在培养的细胞和体内，可能通过BMP 7机制。 （五） （-）纳洛酮：神经变性可通过炎症过程的激活而发生。从过去一年进行的实验中，我们观察到在大鼠大脑中动脉闭塞后一天开始每天给予（+）纳洛酮持续1周时，卒中后行为恢复（身体不对称和神经评分）得到改善。 我们目前正在跟踪这些研究，比较（-）纳洛酮以及检查炎症标志物的变化，这些标志物在中风缺血后上调。