Herbal Effects on Oxidant Dynamics in Cardiomyocytes

描述（由申请人提供）：急性和慢性心脏病，导致显着的发病率和死亡率，往往是由草药治疗的传统医学系统。由于活性氧（reactive oxygen species，ROS）介导心脏病的发病机制，研究抗氧化中药在预防或减轻心脏病中的作用及其机制具有重要意义。我们建议调查两种常用的草药具有抗氧化性能，即葡萄籽原花青素提取物（GSPE）和黄芩提取物（SbE），这是有效的活性氧清除剂。在目前的提案中，我们计划评估是否长期预处理的提取物可以诱导抗氧化保护。我们在鸡心肌细胞预处理的GSPE或SbE或提取物馏分或一些活性成分为48-72小时的初步结果表明显着的保护对内源性和外源性氧化剂冲击。研究结果表明，草药预处理的间接作用降低了细胞内ROS的活性，这将有利于心脏疾病。这可能是通过细胞内抑制ROS的草药代谢物和通过改变基因表达与调节ROS活性的蛋白质合成来实现的。我们推测，GSPE或SbE预处理心肌细胞的结果在显着衰减ROS诱导的损伤和ROS依赖的过程中通过上调细胞的抗氧化能力介导的改变。将在用测试草药预处理1-5天的新生小鼠心肌细胞模型中评价所提出的实验。具体的目的是调查是否预处理GSPE或SbE减弱氧化损伤的心肌细胞在模拟氧化应激，是否草药预处理调节细胞反应的刺激，介导的行动，通过ROS，以及是否改变的反应可以部分解释上调的抗氧化蛋白的基因表达。我们还将评估抗氧化活性的上调是否可以归因于提取物的特定生物活性组分和成分。随后将使用“敲除提取物”确认结果，其中活性成分从其原始基质中去除。这项研究将突出GSPE和SbE在改变心肌细胞内ROS依赖性活动结果中的作用。我们的研究结果可以证明这些草药在预防和治疗心脏病方面具有潜力。拟议的研究将突出GSPE和SbE在改变ROS依赖性细胞事件的结果中的作用。我们的研究结果可以证明草药在预防和治疗心脏病方面的巨大潜力。

DESCRIPTION (provided by applicant): Acute and chronic heart diseases, which cause significant morbidity and mortality, have often been treated by herbal medications in traditional medicinal systems. Since reactive oxygen species (ROS) mediate pathogenesis of heart disease, studying the role of antioxidant herbs in prevention or attenuation of heart disease and the involved mechanisms is extremely important. We propose to investigate two commonly used herbs with antioxidant properties, namely grape seed proanthocyanidin extract (GSPE) and Scutellaria baicalensis extract (SbE), which are potent ROS scavengers. In the current proposal, we plan to evaluate whether long-term pretreatment with the extracts can induce antioxidant protection. Our preliminary results in chick cardiomyocytes pretreated with either GSPE or SbE or extract fractions or some active constituents for 48-72 hr demonstrate significant protection against both an endogenous and exogenous oxidant onslaught. The findings suggest that indirect effects of herbal pretreatment reduce intracellular ROS activity, which would be of benefit in cardiac disease. This is possibly achieved by herbal metabolites within the cell that scavenge ROS and by altered gene expression with synthesis of proteins that modulate ROS activity. We hypothesize that pretreatment of cardiomyocytes with GSPE or SbE results in significant attenuation of ROS-induced injury and alteration in ROS-dependent processes mediated through upregulation of cellular antioxidant capacity. The proposed experiments will be evaluated in a neonatal mouse cardiomyocyte model pretreated with the test herbs for 1-5 days. The specific aims are to investigate whether pretreatment with GSPE or SbE attenuate oxidant injury in the cardiomyocytes during simulated oxidant stress, whether herbal pretreatment modulates cellular responses to stimuli that mediate actions through ROS, and whether the altered responses can be partly explained by upregulated gene expression of antioxidant proteins. We will also evaluate whether the upregulation of antioxidant activity can be attributed to specific bioactive fractions and constituents of the extracts. Results will subsequently be confirmed with the use of "knockout extracts" in which the active constituents are removed from their original matrix. The proposed study will highlight the role of GSPE and SbE in altering the outcome of ROS-dependent activity within cardiomyocytes. Our results could demonstrate that these herbs have a potential in the prevention and treatment of cardiac disease. The proposed study will highlight the role of GSPE and SbE in altering the outcome of ROS-dependent cellular events. Our results could demonstrate a significant potential of the herbs in the prevention and treatment of cardiac disease.