Cerebrovascular Mechanism of Grape Polyphenols-Alcohol

描述(由申请人提供)： 葡萄酒已被证明具有内在的药用特性。多酚类和适量葡萄酒 消费被认为是一种预防药物，但其细胞机制尚不清楚。 鉴于血红素加氧酶(HO)在氧化应激和脑缺血中发挥多种作用，我们验证了HO可能参与多酚神经保护功能的假说。HO酶分解血红素(促氧化剂)形成胆绿素/胆红素、一氧化碳和铁。使用原代神经元培养，我们的结果显示，白藜芦醇是神经元内HO1最有效的诱导剂之一，预处理足以提供神经保护。这项工作，结合乙醇与白藜芦醇或槲皮素具有协同作用的发现，使我们能够提出红酒的一些归属神经保护作用可能是通过诱导HO1以及相关的血红素降解及其生物活性代谢物的有益作用来介导的。我们将确定白藜芦醇或槲皮素单独预处理是否会影响小鼠的血流，以及各自的多酚与酒精联合是否具有协同作用。如果是这样的话，那么将对Hoi-t-小鼠进行测试。其次，我们将确定这些治疗是否可以减少立体定向注射NMDA后纹状体的兴奋性毒性。我们将测试多酚和酒精是否诱导HO1表达的变化，从而导致小鼠神经元培养的细胞存活的变化，以对抗NMDA诱导的毒性。一旦确定了有益的效应，为了解决可能的细胞作用机制，来自HOI-/-小鼠的原代培养的神经元将与WT培养平行进行测试。此外，还将增加HO抑制剂的使用，以进一步检验这一效应的特异性。这些结果将首次表明，预适应是否足以提供神经保护，以及这种作用是否受HOI的调节。我们将检验这一新的假设，即红酒的一些有益作用，如增加脑血流量和减少缺血损伤，可能归因于HO1诱导本身及其生物学作用。这些数据将提出新的途径来解释这种神经保护效应，以及它如何在急性和慢性神经退行性疾病中提供大脑抵抗。

DESCRIPTION (provided by applicant): Wine has been shown to have intrinsic medicinal properties. Polyphenols and moderate wine consumption have been suggested as preventive medicine, but the cellular mechanisms are unclear. Knowing that heme oxygenase (HO) plays various roles in oxidative stress and ischemia, we have tested the hypothesis that HO could participate in polyphenol neuroprotective function. HO enzyme cleaves heme (pro-oxidant) to form biliverdin/bilirubin, carbon monoxide, and iron. Using primary neuronal cultures, our results reveal that resveratrol is one of the most potent inducers of HO1 within neurons and that pre-treatment is sufficient to provide neuroprotection. This work, combined with the finding that ethanol in combination with resveratrol or quercetin would have synergistic effects, allows us to propose that some of the attributed neuroprotective effects of red wine could be mediated through induction of HO1 and the associated beneficial actions of heme degradation and its bioactive metabolites. We will determine whether pre-treatment with resveratrol or quercetin by themselves can affect blood flow and whether the respective polyphenols in combination with alcohol have synergistic effects in mice. If so, then HOI-t- mice will be tested. Second, we will determine whether these treatments can reduce excitotoxicity in the striatum following stereotaxic injection of NMDA. We will test whether polyphenols and alcohol induce changes in HO1 expression that result in changes in cell survival from mouse neuronal cultures against NMDA-induced toxicity. Once a beneficial effect is established, in order to address possible cellular mechanisms of action, primary cultured neurons derived from HOI-/- mice will be tested in parallel with the WT cultures. In addition, use of HO inhibitor will be added to further test the specificity of this effect. These results will indicate for the first time whether preconditioning is sufficient to afford neuroprotection and whether the effect is modulated by HOI. We will test this new hypothesis that some of the beneficial effects attributed to red wine, such as increased cerebral blood flow and reduced ischemic damage, could be attributed to HO1 induction itself and its biological actions. The data will suggest new pathways to explain this neuroprotective effect and how it could provide brain's resistance in acute and chronic neurodegenerative disorders.