Gender Differences in Experimental Aortic Aneurysms

实验性主动脉瘤的性别差异

基本信息

批准号：
8918723
负责人：
Gilbert Rivers Upchurch
金额：
$ 38.91万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8918723
关键词：
Abdominal Aortic Aneurysm Address Adipose tissue Age Agonist Aneurysm Angiotensin II Animals Anti-Inflammatory Agents Anti-inflammatory Aortic Aneurysm Apolipoprotein E Attenuated Binding Biological Response Modifiers Bone Marrow CCL2 gene CD4 Positive T Lymphocytes Caliber Caucasians Cause of Death Cells Cellular Infiltration Chemicals Clinical Collagen Data Development Diet Dinoprostone Disease Elastases Elastin Enzyme-Linked Immunosorbent Assay Enzymes Estradiol Estrogen Receptors Estrogen Therapy Estrogens Evaluation Excision Experimental Models Female Flow Cytometry Gender Gonadal structure Growth Growth Factor HMGB1 Protein Harvest Health Histologic Histology Hormones Human Hypertension Immune Immunohistochemistry In Vitro Incidence Infiltration Inflammation Inflammatory Interferon Type II Interleukin-1 Interleukin-17 Interleukin-6 Knockout Mice Leukocytes Life MMP2 gene MMP9 gene Matrix Metalloproteinases Measurement Measures Mediating Medical Mesenchymal Stem Cells Methods Modeling Molecular Mus Neutrophil Infiltration Nuclear Protein Operative Surgical Procedures Orchiectomy Patients Pattern Perfusion Phytoestrogens Placenta Plasminogen Activator Inhibitor 1 Preventive Production Property Prostaglandins RANTES Regulation Research Proposals Risk Factors Rodent Role Serine Protease Sex Characteristics Smoking History Smooth Muscle Smooth Muscle Actin Staining Method Smooth Muscle Myocytes T-Lymphocyte Tamoxifen Therapeutic Tissues Translating Tumor Necrosis Factor-alpha Tunica Adventitia Up-Regulation Vascular Endothelial Growth Factors Vascular remodeling Western Blotting attenuation base cellular targeting cytokine dietary supplements feeding hormone regulation human female implantation inflammatory marker inhibitor/antagonist interleukin-23 macrophage male mortality mouse model neutrophil paracrine prevent research study treatment strategy vascular inflammation

项目摘要

DESCRIPTION (provided by applicant): Hypothesis: Our previous studies using elastase-perfusion and angiotensin II models of abdominal aortic aneurysms (AAA) have shown that female rodents have a decreased incidence and size of aneurysm formation. We have also recently shown that human placental mesenchymal stem cells (MSCs) are protective in a mouse model of AAA. In the current proposal, we will investigate the role of dietary phytoestrogen as a preventive therapy, and its synergistic effects with MSCs as a treatment strategy to protect against AAA. Methods: We will use an elastase-perfusion and angiotensin II murine model of AAA using male and female wild-type, estrogen receptor knockout mice (ER-α--/- and ER-ß-/-) and ApoE-/- mice. Dietary phytoestrogen will be administered to male and female mice via estrogen-rich or estrogen-free chow. Various subsets of female MSCs (placenta-, bone marrow- or adipose-derived) primed with or without estrogen will be administered intravenously or by aortic implantation in mice models of AAA. Aortic diameter will be measured on day 3, 7 and 14 (elastase perfusion model) and day 28 (angiotensin II model). Aortic tissue will be harvested to analyze pro-inflammatory cytokine (IL-17, TNF-α-, MCP-1, IL-1-ß-, KC and RANTES) and HMGB1 (high mobility group box 1; a pro-inflammatory nuclear protein) production by ELISA, MMP2 and MMP9 activity by zymography, serine proteases (uPA, tPA and PAI-1) by western blots, paracrine factors (VEGF, HGF, PGE2) by ELISA, elastin and collagen degradation as well as aortic smooth muscle expression by histology, and immune cell (macrophages, CD4+ T cells, neutrophils) infiltration by flow cytometry. Results: Preliminary results demonstrate a significant upregulation of estrogen receptor (ER-α) expression in female mice on days 1, 3 and 14 after elastase-perfusion compared to males, as well as in aortic tissue from human females compared to males after AAA. Also, male mice fed a diet rich in estrogen display a significantly decreased aortic diameter, decreased cytokine production (IL-23, IL-1-ß-, IL-6 and IL-27), decreased MMP2 and 9 expression and decreased macrophage and neutrophil infiltration compared to male mice fed an estrogen free diet. Furthermore, treatment with human female MSCs attenuates aortic diameter, pro-inflammatory cytokine production and cell infiltration after AAA. Female MSCs inhibit HMGB1 and IL-17 production more significantly than male MSCs. Also, estradiol-priming of female MSCs offer significantly increased protection from vascular inflammation in aortic tissue from male AAA patients. Conclusions: Dietary estrogen therapy and mesenchymal stem cells can attenuate aneurysm formation and inflammation in the elastase-perfusion murine model of AAA and in human aortic tissue from AAA patients. We propose to delineate the phytoestrogen mediated anti-inflammatory effects, and its crosstalk with various subsets of gender-specific MSCs, on aortic aneurysm formation in the murine (elastase-perfusion and angiotensin II) models as well aortic tissue and cells from AAA patients.

描述（由申请人提供）：假设：我们使用弹性蛋白酶灌注和血管紧张素 II 腹主动脉瘤 (AAA) 模型进行的研究表明，雌性啮齿类动物动脉瘤形成的发生率和大小均较之前有所下降。间充质干细胞 (MSC) 在 AAA 小鼠模型中具有保护作用。在当前的提案中，我们将研究膳食植物雌激素作为激素的作用。预防性治疗及其与 MSC 的协同作用作为预防 AAA 的治疗策略方法：我们将使用雄性和雌性野生型雌激素受体敲除小鼠（ER-α）使用弹性蛋白酶灌注和血管紧张素 II AAA 小鼠模型。 --/- 和 ER-ß-/-) 和 ApoE-/- 小鼠将通过富含雌激素或不含雌激素的食物给予雄性和雌性小鼠。使用或不使用雌激素引发的雌性 MSC 子集（胎盘来源、骨髓来源或脂肪来源）将通过静脉注射或通过主动脉植入 AAA 小鼠模型在第 3、7 和 14 天进行测量（弹性蛋白酶灌注）。模型）和第28天（血管紧张素II模型）将收获主动脉组织以分析促炎细胞因子（IL-17、TNF-α-、通过 ELISA 检测 MCP-1、IL-1-ß-、KC 和 RANTES）和 HMGB1（高迁移率族盒 1；一种促炎核蛋白），通过酶谱法检测 MMP2 和 MMP9 活性，丝氨酸蛋白酶（uPA、tPA 和 PAI） -1) 通过蛋白质印迹，旁分泌因子（VEGF、HGF、PGE2）通过 ELISA、弹性蛋白和胶原蛋白降解以及主动脉平滑肌通过组织学检测肌肉表达，通过流式细胞术检测免疫细胞（巨噬细胞、CD4+ T 细胞、中性粒细胞）浸润结果：初步结果表明，雌性小鼠在术后第 1、3 和 14 天雌激素受体 (ER-α) 表达显着上调。与雄性相比，以及在 AAA 后，与雄性相比，人类女性的主动脉组织中的弹性蛋白酶灌注也显示出，喂食富含雌激素的饮食的雄性小鼠的主动脉显着减少。与喂食无雌激素饮食的雄性小鼠和人类雌性小鼠相比，直径增加，细胞因子产生减少（IL-23、IL-1-ß-、IL-6 和 IL-27），MMP2 和 9 表达减少，巨噬细胞和中性粒细胞浸润减少。 AAA 后，MSC 比雄性 MSC 更显着地减少主动脉直径、促炎细胞因子的产生和细胞浸润。此外，雌二醇引发的雌性 MSC 显着增强了对雄性 AAA 患者主动脉组织血管炎症的保护作用。结论：膳食雌激素治疗和间充质干细胞可以减轻 AAA 弹性蛋白酶灌注小鼠模型和人类的动脉瘤形成和炎症。我们建议描述植物雌激素介导的抗炎作用及其与不同性别亚群的相互作用。 MSC 对小鼠（弹性蛋白酶灌注和血管紧张素 II）模型以及 AAA 患者的主动脉组织和细胞中主动脉瘤形成的影响。