Gender Differences in Experimental Aortic Aneurysms

假设：我们以前的研究使用弹性蛋白酶灌注和血管紧张素II模型的腹主动脉 动脉瘤（AAA）的研究表明，雌性啮齿动物的动脉瘤发生率和大小都有所降低 阵我们最近还表明，人胎盘间充质干细胞（MSC）是保护性的， 在AAA的小鼠模型中。在目前的建议中，我们将研究膳食植物雌激素的作用， 预防性治疗，以及其与MSC的协同作用作为预防AAA的治疗策略。 方法：我们将使用弹性蛋白酶灌注和血管紧张素II的AAA小鼠模型， 野生型雌激素受体敲除小鼠（ER-β-/-和ER-β-/-）和ApoE-/-小鼠。膳食植物雌激素将是 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。女性的各种子集 将施用用或不用雌激素致敏的MSC（胎盘、骨髓或脂肪来源的）。 静脉内或通过主动脉植入在AAA小鼠模型中。将在第3、7天测量主动脉直径 第14天（弹性蛋白酶灌注模型）和第28天（血管紧张素II模型）。将采集主动脉组织进行分析 促炎细胞因子（IL-17、TNF-α、MCP-1、IL-1 β、KC和RANTES）和HMGB 1（高迁移率组 框1;促炎性核蛋白）的产生，MMP 2和MMP 9的活性， 丝氨酸蛋白酶（uPA、tPA和派-1）通过蛋白质印迹，旁分泌因子（VEGF、HGF、PGE 2）通过ELISA， 弹性蛋白和胶原降解以及主动脉平滑肌组织学和免疫细胞表达 （巨噬细胞、CD 4 + T细胞、嗜中性粒细胞）浸润。 结果：初步结果表明，雌激素受体（ER-α）表达的显着上调， 与雄性相比，在弹性蛋白酶灌注后第1、3和14天，雌性小鼠的主动脉组织中， 与AAA后的男性相比。此外，喂食富含雌激素的饮食的雄性小鼠显示出显著的 主动脉直径减小，细胞因子产生减少（IL-23、IL-1、IL-6和IL-27），MMP 2和 9表达和减少的巨噬细胞和中性粒细胞浸润相比，喂食雌激素 免费饮食。此外，用人类雌性MSC治疗可减小主动脉直径，促炎性细胞因子， AAA后细胞因子产生和细胞浸润。雌性MSC抑制HMGB 1和IL-17的产生更多 明显高于男性MSCs。此外，雌二醇引发的雌性MSC提供了显着增加的保护 来自男性AAA患者的主动脉组织中的血管炎症。 结论：饮食雌激素治疗和间充质干细胞可以减少动脉瘤的形成， 在AAA的弹性蛋白酶灌注鼠模型和来自AAA患者的人主动脉组织中的炎症。我们 建议描绘植物雌激素介导的抗炎作用，及其与各种 性别特异性MSC亚群对小鼠主动脉瘤形成的影响（弹性蛋白酶灌注和 血管紧张素II）模型以及来自AAA患者的主动脉组织和细胞。

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-¿-/- and ER-b-/-) 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-¿, MCP-1, IL-1¿, 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-a) 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¿, 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.