The Role of 12-Lipoxygenase in Aldosterone Secretion

12-脂氧合酶在醛固酮分泌中的作用

基本信息

批准号：
8957498
负责人：
Phillip G Kopf
金额：
$ 41.78万
依托单位：
MIDWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8957498
关键词：
12-HETE Acids Adrenal Glands Affinity Aldosterone Alzheimer&apos s Disease Angiotensin II Arachidonate 12-Lipoxygenase Arachidonic Acids Atherosclerosis Binding Calcium Calcium Signaling Cardiovascular Diseases Cell membrane Cells Congestive Heart Failure Corticotropin Cyclic AMP Data Diabetes Mellitus Disease Fostering G-Protein-Coupled Receptors GTP-Binding Proteins Goals Health Heart Hypertrophy Hydroxyeicosatetraenoic Acids Hyperaldosteronism Hyperplasia Hypertension Inflammatory Laboratories Malignant Neoplasms Measures Mediating Membrane Methods Mission Morbidity - disease rate Outcome Parkinson Disease Pathway interactions Patients Phospholipase C Potassium Production Public Health Receptor Activation Receptor, Angiotensin, Type 1 Regulation Research Role Signal Transduction Stimulus Testing Thrombosis Work Zona Glomerulosa adrenic acid analog base functional group guanine nucleotide binding protein improved inhibitor/antagonist innovation liquid chromatography mass spectrometry mortality novel novel therapeutic intervention public health relevance receptor receptor coupling remediation

项目摘要

DESCRIPTION (provided by applicant): Evidence exists for an essential role of the 12-lipoxygenase (12-LO) pathway in Ang II-stimulated aldosterone secretion. However, the identity of 12-LO metabolites, as well as the mechanism and extent by which these metabolites contribute to Ang II-stimulation of aldosterone secretion remains unknown. The long-term goal of these studies is to understand the role of 12-LO in the regulation of aldosterone secretion. The objective of this proposal is to identify the 12-LO metabolites that contribute to Ang II-stimulated aldosterone secretion and to characterize the receptor that mediates the action of these active metabolites. The central hypothesis is that arachidonic acid (AA)-derived hydroxyeicosatetraenoic acids (HETEs) and/or adrenic acid (AdA)-derived hydroxydocosatetraenoic acids (HDTEs) mediate Ang II-stimulated aldosterone secretion by activation of a guanine nucleotide binding protein (G-protein) coupled receptor (GPCR). This hypothesis has been formulated on the basis of preliminary data produced in the applicant's laboratory. The rationale for the proposed research is that identification of the 12-LO metabolites and their receptor that mediates aldosterone secretion would provide a novel pharmacological target for the remediation of circulating aldosterone levels. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Identify the 12-LO metabolites that contribute to aldosterone secretion; and 2) Characterize the receptor that mediates the activity of 12-LO metabolites in ZG cells. Under the first aim, 12-LO AA and AdA metabolites that increase following these steroidogenic stimuli will be identified by liquid chromatography/mass spectrometry (LC/MS), and then these metabolites will be tested for steroidogenic activity. Under the second aim, the ability and potency of these 12-LO AA and AdA metabolites to activate a GPCR on ZG membranes will be tested by GTPγ[35S] binding, as well as their ability to increase cAMP and intracellular calcium. Additionally, the type of Ga subunit that is activated by these metabolites will also be determined. Furthermore, analogs of 12-HETE and 14- HDTE will be used to determine the functional groups necessary for both receptor activation and calcium signaling. The approach is innovative, because it will develop specific LC/MS methods to measure adrenal HETEs and HDTEs and define the role of both AA and AdA metabolites in aldosterone production. The proposed research is significant, because the characterization of a 12(S)-HETE and/or 14(S)-HDTE receptor is an essential step towards understanding the critical role of 12-LO metabolites in the regulation of aldosterone secretion. Ultimately, characterization of this novel GPCR will provide an innovative pharmacological target to not only ameliorate cardiovascular disease associated with hyperaldosteronism, but potentially many other diseases in which 12-LO metabolites have been implicated including inflammatory, hyperproliferative, and hyperplastic diseases such as atherosclerosis, diabetes, Parkinson's disease, Alzheimer's disease, and cancer.

描述（由申请人提供）：有证据表明 12-脂氧合酶 (12-LO) 途径在 Ang II 刺激的醛固酮分泌中发挥重要作用，但 12-LO 代谢物的特性以及其机制和程度尚不清楚。这些代谢物对 Ang II 醛固酮分泌的刺激作用尚不清楚。这些研究的长期目标是了解 12-LO 在调节中的作用。该提案的目的是鉴定有助于 Ang II 刺激醛固酮分泌的 12-LO 代谢物，并表征介导这些活性代谢物作用的受体。中心假设是花生四烯酸 (AA)-。衍生的羟基二十四烯酸（HETE）和/或肾上腺酸（AdA）衍生的羟基二十二碳四烯酸（HDTE）介导血管紧张素通过鸟嘌呤核苷酸结合蛋白（G蛋白）偶联受体（GPCR）的激活II刺激醛固酮分泌。该假设是基于申请人实验室产生的初步数据而提出的。所提出的研究的基本原理是鉴定。 12-LO代谢物及其介导醛固酮分泌的受体的研究将为循环醛固酮水平的修复提供新的药理学靶点，在强有力的初步数据的指导下，这一假设将得到检验。追求两个具体目标：1) 鉴定有助于醛固酮分泌的 12-LO 代谢物；2) 表征 ZG 细胞中介导 12-LO 代谢物活性的受体。这些类固醇生成刺激后增加的物质将通过液相色谱/质谱 (LC/MS) 进行鉴定，然后将在第二步中测试这些代谢物的类固醇生成活性。目的是通过 GTPγ[35S] 结合来测试这些 12-LO AA 和 AdA 代谢物激活 ZG 膜上 GPCR 的能力和效力，以及它们增加 cAMP 和细胞内钙的能力。此外，还测试了 Ga 的类型。此外，还将确定由这些代谢物激活的亚基，12-HETE 和 14-HDTE 的类似物将用于确定受体激活和钙信号传导所需的功能基团。具有创新性，因为它将开发特定的 LC/MS 方法来测量肾上腺 HETE 和 HDTE，并确定 AA 和 AdA 代谢物在醛固酮生产中的作用。这项研究具有重要意义，因为 12(S)-HETE 的表征和/或 14(S)-HDTE 受体是了解 12-LO 代谢物在醛固酮分泌调节中的关键作用的重要一步，最终是这种新型 GPCR 的表征。将提供一个创新的药理学靶点，不仅可以改善与醛固酮增多症相关的心血管疾病，还可以改善与 12-LO 代谢物有关的许多其他疾病，包括炎症、过度增殖和增生性疾病，如动脉粥样硬化、糖尿病、帕金森病、阿尔茨海默病、和癌症。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

M3-subtype muscarinic receptor activation stimulates intracellular calcium oscillations and aldosterone production in human adrenocortical HAC15 cells.

M3 亚型毒蕈碱受体激活刺激人肾上腺皮质 HAC15 细胞的细胞内钙振荡和醛固酮产生。

DOI：
发表时间：
2018
期刊：
影响因子：
4.1
作者：
Malaiyandi, Latha M;Sharthiya, Harsh;Surachaicharn, Nuntida;Shams, Yara;Arshad, Mohammad;Schupbach, Chad;Kopf, Phillip G;Dineley, Kirk E
通讯作者：
Dineley, Kirk E

Adrenal Corticosteroid Perturbation by the Endocrine Disruptor BDE-47 in a Human Adrenocortical Cell Line and Male Rats.

内分泌干扰物 BDE-47 对人肾上腺皮质细胞系和雄性大鼠的肾上腺皮质类固醇干扰。