UKY DENTAL COBRE: ORAL INFECTIONS: COX-2

英国牙科 COBRE：口腔感染：COX-2

• 批准号: 7610651
• 负责人: Charles David Loftin
• 金额: $ 26.95万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610651
• 关键词: Abdomen; Abdominal Aortic Aneurysm; Accounting; Age; Angiotensin II; Aorta; Apolipoprotein E; Arachidonate 15-Lipoxygenase; Arterial Fatty Streak; Atherosclerosis; Attenuated; Bacteremia; Bone Resorption; Caliber; Cardiovascular Diseases; Centers of Research Excellence; Cholesterol; Chronic; Class; Computer Retrieval of Information on Scientific Projects Database; Coupled; Dental; Detection; Disease; Effectiveness; Funding; Genetic; Grant; Incidence; Infection; Inflammation; Inflammatory; Inflammatory Response; Infusion procedures; Institution; Lipoproteins; Localized; Mediator of activation protein; Mus; Numbers; Oral; Oral cavity; Patients; Periodontal Diseases; Periodontitis; Play; Polymerase Chain Reaction; Porphyromonas gingivalis; Procedures; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Protein Isoforms; Research; Research Personnel; Resources; Risk; Role; Serum; Source; Surface; Techniques; Thinking; Time; Tissues; United States National Institutes of Health; Vascular Diseases; Week; alveolar bone; aortic arch; atherogenesis; cyclooxygenase 1; cyclooxygenase 2; day; male; microbial; mouse model; protein expression; trend

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Specific Aim 1. Determine the role of COX-2 in P. gingivalis-induced periodontal disease and atherosclerosis. Periodontitis is a chronic inflammatory disease resulting from microbial infection of the oral cavity. The disease is initiated by gram-negative anaerobic species including Porphyromonas gingivalis. P. gingivalis infection may produce inflammation localized within the oral cavity which results in alveolar bone resorption and may produce a systemic inflammatory response resulting from bacteremia. The systemic inflammatory response is thought to account for the increased risk of developing complications of cardiovascular disease in patients with periodontitis. Prostaglandins are a class of inflammatory mediators that are significantly increased in diseased periodontal tissues. The synthesis of prostaglandins requires the activity of a cyclooxygenase, and cyclooxygenase-2 (COX-2) is the isoform that is highly induced by gram-negative bacterial products and is increased in atherosclerotic lesions. The current studies utilized an established mouse model of P. gingivalis-induced periodontitis to examine the role of COX-2-derived prostaglandins in alveolar bone resorption and atherosclerosis. Alveolar bone resorption and cardiovascular disease resulting from P. gingivalis requires chronic infection to be established in the oral cavity of the mice. Therefore, the mice are inoculated with P. gingivalis 4 days a week, every 3rd week, over a 12 week course. We have optimized the PCR technique to confirm that chronic infection with P. gingivalis is established in the mice. We have also utilized the highly sensitive technique, real-time PCR, to examine the expression of COX-2, as well as downstream prostaglandin synthases in aortic tissue. Furthermore, we have optimized immunohistochemical analysis for detection of COX-2 protein expression in the aortas of mice. These procedures will allow us to examine the effects of P. gingivalis infection on the expression of COX-2 and downstream prostaglandin synthases. Studies are in progress to examine the effectiveness of genetic or pharmacological inactivation of COX-2 in reducing P. gingivalis-induced alveolar bone resorption and atherosclerosis in mice. Specific Aim 2. Define the role of 12/15-lipoxygenase in P. gingivalis-induced periodontal disease and atherosclerosis. Angiotensin II (AngII) markedly accelerates atherogenesis in hyperlipidemic mice. Moreover, AngII increases the expression of 12/15-lipoxygenase (12/15-LO). Deficiency of 12/15-LO markedly attenuates atherosclerosis in hyperlipidemic mice. Therefore, we sought to determine if 12/15-LO plays a role in AngII-induced vascular disease. Age matched male apolipoproteinE deficient (apoE-/-) mice or 12/15-LO deficient apoE-/- mice were infused with AngII (1,000 ng/kg/min) for 28 days. Total serum cholesterol or lipoprotein profile was not altered in 12/15-LO deficient mice following AngII infusion compared to control. 12/15-LO modestly decreased AngII-induced atherosclerotic lesion formation (12/15-LO+/+: 1.26% vs 12/15-LO-/-: 1.99%; P = 0.12) on the intimal surface of the aortic arch however, this decrease did not reach statistical significance. Additionally, 12/15-LO deficiency decreased the incidence of AngII-induced abdominal aortic aneurysm formation (12/15-LO+/+: 42% vs 12/15-LO-/-: 17%; P = 0.12) and the abdominal aortic diameter (12/15-LO+/+: 1.6 mm vs 12/15-LO-/-: 1.1 mm; P = 0.11), however, the reduction in both of these parameters did not reach statistical significance. While these results suggest 12/15-LO does not significantly alter AngII-induced atherosclerosis and AAA formation, the number of mice utilized in these studies did not result in a power of 0.08. This coupled with the trend toward a decrease in both atherosclerosis and AAA formation suggest that additional studies are required to define the effect of 12/15-LO on AngII-induced vascular disease.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 具体目的1。确定COX-2在牙龈疟原虫诱导的牙周疾病和动脉粥样硬化中的作用。 牙周炎是由口腔微生物感染引起的慢性炎症性疾病。该疾病是由包括牙龈卟啉单胞菌在内的革兰氏阴性厌氧物种引发的。牙龈疟原虫感染可能会导致口腔内局部炎症，从而导致肺泡骨吸收，并可能引起菌血症引起的全身性炎症反应。人们认为系统性炎症反应是牙周炎患者患心血管疾病并发症的风险增加的。前列腺素是一类炎症介质，在患病的牙周组织中显着增加。前列腺素的合成需要环氧酶的活性，而环氧酶-2（COX-2）是由革兰氏阴性细菌产物高度诱导的同工型，并且在动脉粥样硬化病变中增加。 当前的研究利用了牙龈疟原虫诱导的牙周炎的既定小鼠模型来检查COX-2衍生的前列腺素在肺泡骨吸收和动脉粥样硬化中的作用。牙龈疟原虫引起的肺泡骨吸收和心血管疾病需要在小鼠的口腔中建立慢性感染。因此，在12周的课程中，每周4天，每3周，每周4天4天接种牙龈疟原虫。我们已经优化了PCR技术，以确认在小鼠中建立了牙龈假单胞菌的慢性感染。我们还利用了高度敏感的技术，实时PCR来检查COX-2的表达以及主动脉组织中的下游前列腺素合成酶。此外，我们已经优化了免疫组织化学分析，用于检测小鼠主动脉中COX-2蛋白的表达。这些程序将使我们能够检查牙龈疟原虫感染对COX-2和下游前列腺素合成酶表达的影响。研究正在进行研究，以研究COX-2在减少牙龈疟原虫诱导的肺泡骨吸收和动脉粥样硬化中的遗传学或药理失活的有效性。 具体目标2。定义12/15-脂氧酶在牙龈疟原虫诱导的牙周疾病和动脉粥样硬化中的作用。 血管紧张素II（AngII）显着加速了高脂小鼠的动脉粥样硬化。 此外，AngII增加了12/15-脂氧酶（12/15-LO）的表达。 12/15 lo的缺乏显着减弱高脂小鼠的动脉粥样硬化。 因此，我们试图确定12/15-LO是否在Angii诱导的血管疾病中起作用。 年龄匹配的雄性载脂蛋白缺乏蛋白（APOE - / - ）小鼠或12/15-lo缺乏的APOE-/ - 小鼠注入ANGII（1,000 ng/kg/min）28天。 与对照相比，在ANGII输注后，12/15-LO缺乏小鼠的总血清胆固醇或脂蛋白谱并未改变。 12/15-LO适度降低的Angii诱导的动脉粥样硬化病变形成（12/15-LO+/+：1.26％vs 12/15-LO - / - ：1.99％; P = 0.12; P = 0.12）在主动脉弓的内膜表面上，但是这种降低的降低尚未达到统计学意义。 此外，12/15-lo缺乏症降低了血管诱导的腹主动脉瘤形成的发生率（12/15-lo+/+：42％vs 12/15-lo-/ - / - ：17％; p = 0.12）和腹部主动脉直径（12/15-lo+/++/++/++/++++/+lo）。这两个参数的降低均未达到统计意义。 尽管这些结果表明12/15-LO并未显着改变Angii诱导的动脉粥样硬化和AAA形成，但这些研究中使用的小鼠数量并未导致0.08的功率。 结合降低动脉粥样硬化和AAA形成的趋势表明，需要其他研究来定义12/15-LO对Angii诱导的血管疾病的影响。