权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Role Of Neuropeptides And Biogenic Amines In Stress and Brain Inflammation

神经肽和生物胺在压力和脑炎症中的作用

基本信息

批准号：
7735135
负责人：
JUAN M SAAVEDRA
金额：
$ 230.13万
依托单位：
NATIONAL INSTITUTE OF MENTAL HEALTH
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7735135
关键词：
AGTR2 gene Acute Adrenal Cortex Hormones Adrenal Glands Agonist Aldosterone Angiotensin II Angiotensin II Receptor Angiotensin Receptor Animal Model Animals Anti-Inflammatory Agents Anti-inflammatory Anxiety Area Binding Biogenic Amines Biological Psychiatry Biotin Birth Brain Brain Diseases CD14 Antigen CD14 gene CGP-42112 Cerebrum Characteristics Chronic Class Clinical Protocols Collaborations Complex Condition Corticosterone Corticotropin Corticotropin-Releasing Hormone Coupling Cultured Cells Development Development, Other Dinoprostone Disease Dissection Encephalitis Endothelial Cells Endothelium Endotoxins Equilibrium FOS gene Fright Gene Deletion Gene Expression Gene Silencing Genes Genetic Models Genetic Transcription Goals High Blood Pressure Hippocampus (Brain)Hormones Human Human Volunteers Hypertension Hypothalamic structure Immune Immune response Immunohistochemistry In Vitro Inflammation Inflammatory Inflammatory Response Label Lead Limbic System Lipopolysaccharides Localized Mediating Mental Depression Microglia Mineralocorticoids Mus NADP NF-kappa B National Institute of Mental Health Nerve Degeneration Neurodegenerative Disorders Neurons Neuropeptides Nitric Oxide Nitric Oxide Synthase Numbers Oxidases Pathogenesis Pathway interactions Peripheral Pharmaceutical Preparations Phenotype Pituitary Gland Play Post-Traumatic Stress Disorders Prevention approach Production Property Proteins Proteomics Purpose Rattus Reaction Reactive Oxygen Species Receptor Gene Regulation Reporting Research Rodent Rodent Model Role Signal Transduction Site Spleen Stimulus Streptavidin Stress System Techniques Therapeutic Time Transcriptional Activation Transfection Treatment Efficacy Tyrosine 3-Monooxygenase Up-Regulation Work adiponectin base candesartan cell type cerebrovascular cyclooxygenase 2 cytokine familial hypertension hypothalamic-pituitary-adrenal axis in vivo laser capture microdissection locus ceruleus structure macrophage molecular recognition monocyte mouse model multitask neuropsychiatry novel novel strategies novel therapeutics paraventricular nucleus prevent protein protein interaction receptor research study response response to injury transcription factor

项目摘要

Brain inflammation. In previous work, we had found that sustained in vivo AT1 receptor blockade with the ARB candesartan reversed, in a rat model of genetic hypertension, the cerebrovascular inflammation which is characteristic of hypertension in both rodents and humans. We asked the question whether ARBs could reduce other types of brain inflammation. During FY 2008, we focused on the effect of ARBs on the rat innate immune response, produced by systemic administration of the bacterial endotoxin lipopolysaccharide (LPS). We found that ARBs limited the peripheral and brain inflammatory responses to systemic immune challenge in the rat. In vivo, ARBs reduced the LPS-induced inflammatory reaction in rodent spleen, adrenal gland, pituitary gland, and in multiple brain areas including the hypothalamus, olfactory system, hippocampus and cortex. Anti-inflammatory effects of ARBs were more pronounced in areas expressing large numbers of neuronal AT1 receptors. In addition, ARBs reduced the LPS-induced increase of the pro-inflammatory mineralocorticoid hormone aldosterone and enhanced the production of the anti-inflammatory hormone adiponectin. Moreover, ARBs rapidly reduced, in vitro, the LPS-induced pro-inflammatory gene expression and secretion of pro-inflammatory cytokines in cultured human monocytes. We next focused on the elucidation of the mechanisms of the anti-inflammatory effects of ARBs, and we asked the question which of the major mechanisms of inflammation stimulated by LPS were reduced or eliminated by ARB administration. We found that ARBs reduced, in vivo, the LPS-induced production of multiple pro-inflammatory factors, including reactive oxygen species, nitric oxide and prostaglandin E2. This implicates mechanisms involving Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidase, cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS) and transcription factors such as NF-kappaB. We found similar ARB effects, in vitro, in cultured human circulating monocytes. A repeated finding after ARB administration was the decrease in the constitutive and in the LPS-induced expression of the LPS receptor CD14. We hypothesize that interference with the LPS molecular recognition complex may explain part of the anti-inflammatory effects of ARBs. We are now focusing on the further clarification of the mechanisms of action of ARBs. We use a mouse model of AT1 receptor gene-deletion, to determine the effect of LPS when AT1 receptors are not expressed from birth. Cerebral endothelium plays a crucial role in the propagation of inflammatory signals throughout the brain and is critical for activation of specific groups of neurons involved in central response to systemic inflammation. We use cultures of cerebral endothelial cells and isolated microglia to study the relationship between LPS and AT1 receptors, to determine cellular colocalization and direct protein-protein interactions. Studies include gene transfections and gene silencing techniques in phenotype rescue studies and to identify critical components mediating the effect of ARBs in particular cell types. These experiments will further clarify the mechanisms of the anti-inflammatory effects of ARBs. In previous work, we have reported that CGP42112, an agonist of a second type of Ang II receptors, the AT2 receptors, actively inhibits the LPS-induced inflammation in human monocytes. During FY 2008 we discovered that, in rodents, CGP42112 effectively decreases LPS-induced inflammation in vivo. The anti-inflammatory effects of CGP42112 could be the result of AT2 receptor stimulation and/or binding to a novel macrophage receptor that we have previously discovered and reported, localized to brain and circulating macrophages. This novel non-Angiotensin receptor has not yet been characterized. In collaboration with Drs. Markey, NIMH, we are attempting to identify proteins which bind to CGP42112 using biotin-labeled CGP42112 and isolation by streptavidin coupling, and by laser capture microdissection, immunohistochemistry and proteomics. In other studies we will characterize the mechanisms of CGP42112 novel anti-inflammatory effects using AT2 gene-deleted mice and human circulating monocytes. Stress. We previously demonstrated that orally administered ARBs limited the HPA axis, sympathetic and cortical responses to isolation stress, and that these compounds reduced anxiety in rodents. On this basis, we previously proposed the first clinical protocol to evaluate the effects of AT1 receptor antagonists in the fear-startle response in human volunteers, with the goal to determine if AT1 receptor antagonists are effective in reducing anxiety and stress in humans. During FY 2008, we further explored the anti-stress effects of ARBs in response to immune challenge. We now find that ARBs limit the HPA axis response to the endotoxin lipopolysaccharide (LPS) in the adrenal gland, pituitary and hypothalamus. ARBs decreased the LPS-induced upregulation of the early transcription factor c-fos and microglial activation in the paraventricular nucleus. While eliminating the LPS-induced up-regulation of the hormone aldosterone, ARBs do not decrease LPS-induced ACTH and corticosterone responses, demonstrating that ARBs do not limit the anti-inflammatory effects of LPS-induced corticosteroid release. We found that the effect of ARBs on the HPA axis depend on the type of stress. During isolation stress, ARBs prevent HPA axis activation, but they do not prevent the HPA axis response to inflammation. We have also found that ARBs prevent the stress-induced increase in brain sympathetic activity by suppressing the stress-induced tyrosine hydroxylase gene transcription in the rat locus coeruleus. Our experiments are now focused on the elucidation of the mechanisms of the ARB control of tyrosine hydroxylase transcription in the locus coeruleus, the regulation of corticotrophin-releasing factor in the paraventricular nucleus, and the determination of additional brain sites involved on the anti-stress effects of ARBs. To this end we combine selective dissection techniques, gene microarray and proteomic studies. In conclusion, we demonstrated during the FY 2008 that ARBs may be considered as a novel class of multitasking anti-stress, anti-anxiety, anti-inflammatory medications in the treatment of brain disorders. They are already widely used to treat high blood pressure in humans and are safe and devoid of addictive properties, Elucidation of their mechanisms of action may lead to the development of other compounds of great therapeutic potential.

脑部炎症。在之前的工作中，我们发现，在遗传性高血压大鼠模型中，使用 ARB 坎地沙坦持续体内 AT1 受体阻断可逆转脑血管炎症，而脑血管炎症是啮齿类动物和人类高血压的特征。我们提出了这样的问题：ARB 是否可以减少其他类型的脑部炎症。 2008 财年，我们重点研究了 ARB 对大鼠先天免疫反应的影响，该反应是通过全身施用细菌内毒素脂多糖 (LPS) 产生的。我们发现 ARB 限制了大鼠对全身免疫挑战的外周和大脑炎症反应。在体内，ARB 可以减少啮齿动物脾脏、肾上腺、垂体以及下丘脑、嗅觉系统、海马和皮质等多个大脑区域中 LPS 诱导的炎症反应。 ARB 的抗炎作用在表达大量神经元 AT1 受体的区域更为明显。此外，ARB 还可以减少 LPS 诱导的促炎盐皮质激素醛固酮的增加，并增强抗炎激素脂联素的产生。此外，在体外培养的人单核细胞中，ARB 迅速减少 LPS 诱导的促炎基因表达和促炎细胞因子的分泌。接下来，我们重点阐述了 ARB 的抗炎作用机制，并提出了以下问题：LPS 刺激的炎症的主要机制中，哪些通过 ARB 给药而减轻或消除。我们发现，ARB 在体内减少了 LPS 诱导的多种促炎因子的产生，包括活性氧、一氧化氮和前列腺素 E2。这暗示涉及烟酰胺腺嘌呤二核苷酸磷酸 (NADPH) 氧化酶、环氧合酶-2 (COX-2)、一氧化氮合酶 (NOS) 和转录因子（如 NF-kappaB）的机制。我们在体外培养的人循环单核细胞中发现了类似的 ARB 效应。施用 ARB 后反复发现 LPS 受体 CD14 的组成型表达和 LPS 诱导的表达降低。我们假设干扰 LPS 分子识别复合物可以解释 ARB 的部分抗炎作用。我们现在的重点是进一步阐明ARB的作用机制。我们使用 AT1 受体基因缺失的小鼠模型来确定当 AT1 受体从出生时不表达时 LPS 的影响。脑内皮细胞在炎症信号在整个大脑中的传播中起着至关重要的作用，并且对于激活参与全身炎症中枢反应的特定神经元组至关重要。我们使用脑内皮细胞和分离的小胶质细胞的培养物来研究 LPS 和 AT1 受体之间的关系，以确定细胞共定位和直接的蛋白质-蛋白质相互作用。研究包括表型拯救研究中的基因转染和基因沉默技术，以及鉴定介导 ARB 在特定细胞类型中作用的关键成分。这些实验将进一步阐明ARB的抗炎作用机制。在之前的工作中，我们报道了 CGP42112，第二类 Ang II 受体 AT2 受体的激动剂，可主动抑制 LPS 诱导的人类单核细胞炎症。 2008 财年，我们发现，在啮齿类动物中，CGP42112 能有效减少 LPS 诱导的体内炎症。 CGP42112 的抗炎作用可能是 AT2 受体刺激和/或与我们之前发现和报道的新型巨噬细胞受体结合的结果，该受体定位于大脑和循环巨噬细胞。这种新型非血管紧张素受体尚未得到表征。与博士合作。 Markey，NIMH，我们正在尝试使用生物素标记的 CGP42112 并通过链霉亲和素偶联进行分离，以及通过激光捕获显微切割、免疫组织化学和蛋白质组学来鉴定与 CGP42112 结合的蛋白质。在其他研究中，我们将使用 AT2 基因缺失小鼠和人类循环单核细胞来表征 CGP42112 新型抗炎作用的机制。压力。我们之前证明，口服 ARB 可以限制 HPA 轴、交感神经和皮质对隔离应激的反应，并且这些化合物可以减少啮齿类动物的焦虑。在此基础上，我们之前提出了第一个临床方案来评估AT1受体拮抗剂对人类志愿者恐惧惊吓反应的影响，目的是确定AT1受体拮抗剂是否能有效减轻人类的焦虑和压力。 2008财年，我们进一步探索了ARB针对免疫挑战的抗应激作用。我们现在发现 ARB 限制了肾上腺、垂体和下丘脑中 HPA 轴对内毒素脂多糖 (LPS) 的反应。 ARB 降低了 LPS 诱导的早期转录因子 c-fos 的上调和室旁核中小胶质细胞的激活。在消除 LPS 诱导的醛固酮激素上调的同时，ARB 不会降低 LPS 诱导的 ACTH 和皮质酮反应，这表明 ARB 不会限制 LPS 诱导的皮质类固醇释放的抗炎作用。我们发现 ARB 对 HPA 轴的影响取决于压力的类型。在隔离应激期间，ARB 会阻止 HPA 轴激活，但不会阻止 HPA 轴对炎症的反应。我们还发现，ARB 通过抑制大鼠蓝斑中应激诱导的酪氨酸羟化酶基因转录来防止应激诱导的大脑交感神经活动增加。我们的实验现在集中于阐明ARB控制蓝斑中酪氨酸羟化酶转录的机制、室旁核中促肾上腺皮质激素释放因子的调节，以及确定涉及ARB抗应激作用的其他脑位点。为此，我们结合了选择性解剖技术、基因微阵列和蛋白质组学研究。总之，我们在 2008 财年证明，ARB 可被视为治疗脑部疾病的一类新型多任务抗压力、抗焦虑、抗炎药物。它们已广泛用于治疗人类高血压，并且安全且无成瘾性。阐明其作用机制可能会导致开发其他具有巨大治疗潜力的化合物。

项目成果

期刊论文数量（47）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The future of Latin American science.

拉丁美洲科学的未来。

DOI：
10.1023/a:1021853625299
发表时间：
2002
期刊：
Cellular and molecular neurobiology
影响因子：
4
作者：
Saavedra,JuanM
通讯作者：
Saavedra,JuanM

Increased AT(1) receptor expression and mRNA in kidney glomeruli of AT(2) receptor gene-disrupted mice.

AT(2) 受体基因破坏小鼠肾小球中 AT(1) 受体表达和 mRNA 增加。

DOI：
10.1152/ajprenal.2001.280.1.f71
发表时间：
2001
期刊：
American journal of physiology. Renal physiology
影响因子：
0
作者：
Saavedra,JM;Hauser,W;Ciuffo,G;Egidy,G;Hoe,KL;Johren,O;Sembonmatsu,T;Inagami,T;Armando,I
通讯作者：
Armando,I

Intracisternal administration of Angiotensin II AT1 receptor antisense oligodeoxynucleotides protects against cerebral ischemia in spontaneously hypertensive rats.

血管紧张素 II AT1 受体反义寡脱氧核苷酸的脑池内给药可预防自发性高血压大鼠的脑缺血。