REGULATION OF CORONARY ARTERY FLOW BY ENDOTHELIAL CELLS

描述：心血管疾病仍然是两人的头号杀手。 和美国的女性。到目前为止，研究工作已经针对 了解冠状动脉疾病的发展及其在 心肌缺血、损伤和梗死，这是由于 冠状动脉血流。研究人员将试图描述 导致冠状动脉血流调节的机制， 正常的生理状态，并可能最终提供的潜力， 在受损的心血管状态期间增加冠状动脉血流量。 具体来说，他们将研究细胞外介质， 参与了冠状动脉血流的调节。调解员将 检查的是腺苷嘌呤（即，ATP、ADP和腺苷）。是 假设心脏内皮细胞可以酶促生成 细胞外ATP作为自分泌或旁分泌在血管中起作用 激素.这项研究将：1）确定人类冠状动脉是否 内皮细胞可以酶促产生细胞外ATP; 2）表征 与人冠状动脉质膜相关的胞外激酶 内皮细胞;和3）抑制与内皮细胞相关的胞外激酶的功能， 与人冠状动脉内皮细胞的质膜结合。的 研究人员将使用从人冠状动脉内皮细胞获得的人冠状动脉内皮细胞， 杜克综合癌症中心细胞培养设施，获得了他们的 来自Clonetics或ATCC的细胞。将通过HPLC测量ATP的生成 放射性测定和荧光光谱。蛋白质测定和 鉴定将伴随SDS-PAGE和Western印迹技术。 反义寡核苷酸将用于抑制反义寡核苷酸的功能。 胞外激酶。将使用Student t检验分析获得的数据 重复测量方差分析。该结果将成为未来体内研究的基础。 研究和干预研究。

DESCRIPTION: Cardiovascular disease remains the number one killer of both men and women in the United States. To date, research efforts have been directed at understanding the development of coronary artery disease and its role in myocardial ischemia, injury, and infarction, which are due to alterations in coronary blood flow. The investigators will attempt to characterize the mechanism that results in the regulation of coronary artery blood flow in normal physiological states and may eventually provide the potential for increasing coronary blood flow during compromised cardiovascular states. Specifically, they will investigate the extracellular mediators that may be involved in this regulation of coronary artery blood flow. The mediators to be examined are the adenyl purines (i.e., ATP, ADP, and adenosine). It is hypothesized that the cardiac endothelial cells can enzymatically generate extracellular ATP to act in the blood vessel as an autocrine or paracrine hormone. The proposed research will: 1) determine whether human coronary artery endothelial cells can enzymatically generate extracellular ATP; 2) characterize the ecto-kinase(s) associated with the plasma membrane of human coronary artery endothelial cells; and 3) inhibit the function of the ecto-kinase(s) associated with the plasma membrane of human coronary artery endothelial cells. The investigators will use human coronary artery endothelial cells obtained from the Duke Comprehensive Cancer Center Cell Culture Facility, which obtain their cells from Clonetics or ATCC. The generation of ATP will be measured by HPLC radioactive determination and chemiluminesecence. Protein determination and identification will be accompanied with SDS-PAGE and Western blot technique. Antisense oligonucleotides will be utilized to inhibit the function of the ecto-kinase(s). The data obtained will be analyzed using the Student's t-test and repeated measures ANOVA. The results will form the basis for future in vivo studies and intervention research.