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通过激活载脂蛋白E缺陷小鼠的盐皮质激素受体诱导动脉粥样硬化斑块形成。

Induction of Atherosclerotic Plaques Through Activation of Mineralocorticoid Receptors in Apolipoprotein E-deficient Mice.

作者信息

Marzolla Vincenzo, Armani Andrea, Mammi Caterina, Feraco Alessandra, Caprio Massimiliano

机构信息

Laboratory of Cardiovascular Endocrinology, IRCCS San Raffaele Pisana, Rome, Italy.

Laboratory of Cardiovascular Endocrinology, IRCCS San Raffaele Pisana, Rome, Italy; Department of Human Sciences and Promotion of the Quality of Life, San Raffaele Roma Open University, Rome, Italy;

出版信息

J Vis Exp. 2018 Sep 26(139):58303. doi: 10.3791/58303.

Abstract

Atherosclerosis is due to a chronic inflammatory response affecting vascular endothelium and is promoted by several factors such as hypertension, dyslipidemia, and diabetes. To date, there is evidence to support a role for circulating aldosterone as a risk factor for the development of cardiovascular disease. Transgenic mouse models have been generated to study cellular and molecular processes leading to atherosclerosis. In this manuscript, we describe a protocol that takes advantage of continuous infusion of aldosterone in ApoE mice and generates atherosclerotic plaques in the aortic root after 4 weeks of treatment. We, therefore, illustrate a method for quantification and characterization of atherosclerotic lesions at the aortic root level. The added value of aldosterone infusion is represented by the generation of atherosclerotic lesions rich in lipid and inflammatory cells after 4 weeks of treatment. We describe in detail the staining procedures to quantify lipid and macrophage content within the plaque. Notably, in this protocol, we perform heart tissue-embedding in OCT in order to preserve the antigenicity of cardiac tissue and facilitate detectability of antigens of interest. Analysis of the plaque phenotype represents a valid approach to study the pathophysiology of atherosclerosis development and to identify novel pharmacological targets for the development of anti-atherogenic drugs.

摘要

动脉粥样硬化是由于影响血管内皮的慢性炎症反应所致,并由高血压、血脂异常和糖尿病等多种因素促成。迄今为止,有证据支持循环醛固酮作为心血管疾病发生的危险因素发挥作用。已构建转基因小鼠模型来研究导致动脉粥样硬化的细胞和分子过程。在本手稿中,我们描述了一种方案,该方案利用对载脂蛋白E(ApoE)小鼠持续输注醛固酮,并在治疗4周后在主动脉根部生成动脉粥样硬化斑块。因此,我们阐述了一种在主动脉根部水平定量和表征动脉粥样硬化病变的方法。醛固酮输注的附加价值表现为在治疗4周后生成富含脂质和炎症细胞的动脉粥样硬化病变。我们详细描述了量化斑块内脂质和巨噬细胞含量的染色程序。值得注意的是,在本方案中,我们在最佳切割温度(OCT)化合物中进行心脏组织包埋,以保留心脏组织的抗原性并便于检测感兴趣的抗原。对斑块表型的分析是研究动脉粥样硬化发生的病理生理学以及确定抗动脉粥样硬化药物开发新的药理学靶点的有效方法。

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