Rodrigo Manoj C, Martin Douglas S, Redetzke Rebecca A, Eyster Kathleen M
Hypertension Unit, Division of Basic Biomedical Sciences, University of South Dakota School of Medicine, 414 East Clark Street, Vermillion, SD 57069, USA.
J Pharmacol Toxicol Methods. 2002 Mar-Apr;47(2):87-92. doi: 10.1016/s1056-8719(02)00218-6.
The mesenteric vasculature plays an important role in cardiovascular homeostasis, both in terms of arterial resistance and venous capacitance. However, the limited quantity of tissue available and the activity of RNases and proteases during dissection of the vessels make it difficult to study gene and protein expression in this tissue.
The mesenteric arcade was harvested from rats. The mesenteric arteries and veins were separated from each other and from contaminating adipose, nerve, and connective tissue by blunt dissection while submerged in RNAlater (Ambion). Total ribonucleic acid (RNA) and protein were extracted from the same mesenteric artery and vein samples.
Good quality RNA and protein were obtained by this method. The method preserved the content of proteins including biglycan, adrenergic receptors, estrogen receptors, and protein kinase Cdelta, and also preserved the differential expression of estrogen receptors.
A method for the isolation of high-quality RNA and protein from the arteries and veins of the same mesenteric sample is described. The content of proteins from a variety of cellular compartments and molecular weights was preserved by this method, as well as differential expression of proteins. This method will facilitate the study of gene and protein expression in the mesenteric vasculature in response to pharmacologic treatments and in disease states.
肠系膜血管系统在心血管稳态中起着重要作用，无论是在动脉阻力还是静脉容量方面。然而，可用组织数量有限以及在血管解剖过程中核糖核酸酶和蛋白酶的活性使得研究该组织中的基因和蛋白质表达变得困难。
从大鼠身上采集肠系膜弓。在浸没于RNA Later（安必昂公司）的同时，通过钝性分离将肠系膜动脉和静脉彼此分离，并与污染的脂肪、神经和结缔组织分离。从相同的肠系膜动脉和静脉样本中提取总核糖核酸（RNA）和蛋白质。
通过该方法获得了高质量的RNA和蛋白质。该方法保留了包括双糖链蛋白聚糖、肾上腺素能受体、雌激素受体和蛋白激酶Cδ在内的蛋白质含量，还保留了雌激素受体的差异表达。
描述了一种从同一肠系膜样本的动脉和静脉中分离高质量RNA和蛋白质的方法。该方法保留了来自各种细胞区室和分子量的蛋白质含量以及蛋白质的差异表达。这种方法将有助于研究肠系膜血管系统在药物治疗反应和疾病状态下的基因和蛋白质表达。
