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) 安非他明在神经再生中的作用：通过中风大鼠的身体不对称性和贝德森评分来测量，AM 治疗后显着减少了神经功能缺损。 T2WI和弥散张量成像（DTI）分别用于在卒中后第2、10和25天治疗前和治疗后检查梗死面积和轴突再神经支配。 AM治疗减少了第10天和第25天的组织损失量。重复AM给药后，同侧皮层的各向异性分数显着增加，表明病变侧皮层的轴突生长可能增加。 Western分析表明AM显着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) 的酶活性，这是通过受损侧皮质中的 MMP 酶谱测定得出的。 qRT-PCR用于检测第1次和第2次AM或盐水注射后营养因子的表达。 AM 显着增强了病变侧皮层中 BDNF 的表达，但 BMP7 或 CART 没有显着增强。总之，中风后使用 AM 治疗有利于行为恢复，这与分数各向异性活性的增加、纤维束成像中纤维生长的增强、突触发生、BDNF 的上调以及主要在病变皮质中的 MMP 活性有关。 我们的数据表明，AM 治疗后，同侧皮质可能是中风大脑的主要作用目标。 (2) 腺苷 A3R 受体：我们报道，高剂量的冰毒抑制成年雄性 A3R 无效突变 (-/-) 小鼠及其对照 (+/+) 小鼠的运动活性，在 -/- 小鼠中发现抑制程度更大。注射冰毒或盐水后 3 天收集脑组织。 通过 HPLC 检查，冰毒治疗降低了 +/+ 和 -/- 小鼠的纹状体多巴胺 (DA) 水平，仅在冰毒治疗后 -/- 动物中发现 DOPAC/DA 比率增加。与+/+小鼠相比，Meth还显着增加了-/-小鼠纹状体中的离子钙结合接头分子1 (Iba-1)和裂解的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 VMAT2 的药理抑制会通过增加细胞质 DA 和炎症来增强冰毒毒性。与+/+小鼠相比，-/-小鼠中纹状体VMAT2表达显着降低，这表明-/-小鼠对冰毒损伤敏感性的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 -/- 小鼠对高剂量的冰毒更敏感。 (3)pifithrin-a (PFT-a)：(A) 我们发现 PFTa 对多巴胺能神经元中的 Meth 介导的损伤具有保护作用。高剂量的冰毒降低了原代多巴胺能神经元培养物中的 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行性变化。我们的数据表明，用 p53 抑制剂治疗或抑制 p53 表达可限制因暴露于多巴胺能神经毒素而引起的神经元功能障碍和细胞死亡。 (B) PFT 的全身施用增强了体内多巴胺能神经元移植物的存活。 PFT 治疗可抑制细胞死亡，增加 TH (+) 细胞的存活率，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 p53 的激活是损伤后或神经修复过程中细胞死亡的重要介质。 短暂抑制 p53 可能会增加脑损伤后神经元或 NPC 的存活率。 (4) 视黄酸(RA)：用9顺式RA(9cRA)预处理可增加中风大鼠的运动活性，减轻神经功能缺损，并减少脑梗塞和TUNEL标记。 9cRA 增加大脑中骨形态发生蛋白 7 (BMP7) 的表达。 9cRA 的保护反应可以被 BMP 拮抗剂 noggin 拮抗。我们的数据表明，RA 可能通过 BMP7 机制在培养细胞和体内诱导针对冰毒毒性的保护性反应。 (5) (-)纳洛酮：神经变性可通过炎症过程的激活而发生。从去年进行的实验中，我们观察到，当大鼠大脑中动脉闭塞后一天开始连续 1 周每天给予 (+) 纳洛酮时，中风后的行为恢复（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，并检查中风缺血后炎症标志物的变化。