Seiffert D, Crain K, Wagner N V, Loskutoff D J
Department of Vascular Biology, Scripps Research Institute, La Jolla, California 92037.
J Biol Chem. 1994 Aug 5;269(31):19836-42.
A competitive polymerase chain reaction (PCR) assay was developed to quantitate vitronectin (Vn) mRNA in murine tissues using a synthetic RNA as an external standard. Although the liver contained the highest concentration of Vn mRNA, significant levels were also detected in the brain (25-fold less) and in adipose tissue, heart, and skeletal muscle (100-fold less than liver). Lower concentrations also were detected in the lung, uterus, testis, and thymus, and little or no Vn mRNA could be detected in kidney, spleen, and blood. These results indicate that significant amounts of Vn mRNA are produced in extrahepatic organs. The regulation of Vn gene expression in vivo was studied in a murine model system in which acute systemic inflammation was induced by endotoxin administration. Plasma Vn levels increased 2- to 3-fold within 16 h after endotoxin administration and remained elevated for up to 72 h. This increase appeared to result from increased synthesis in the liver since the steady-state level of hepatic Vn mRNA increased 4-fold after endotoxin administration. Moreover, Vn mRNA levels in heart, lung, and brain were not significantly increased by endotoxin. These results suggest that Vn gene expression in vivo is regulated in a tissue-specific manner and identify Vn as a novel acute phase reactant.
开发了一种竞争性聚合酶链反应(PCR)检测方法,以使用合成RNA作为外标来定量小鼠组织中的玻连蛋白(Vn)mRNA。尽管肝脏中Vn mRNA的浓度最高,但在大脑中也检测到显著水平(比肝脏低25倍),在脂肪组织、心脏和骨骼肌中也检测到显著水平(比肝脏低100倍)。在肺、子宫、睾丸和胸腺中也检测到较低浓度,而在肾脏、脾脏和血液中几乎检测不到或检测不到Vn mRNA。这些结果表明,肝外器官中产生了大量的Vn mRNA。在通过给予内毒素诱导急性全身炎症的小鼠模型系统中研究了体内Vn基因表达的调控。给予内毒素后16小时内,血浆Vn水平增加2至3倍,并在长达72小时内保持升高。这种增加似乎是由于肝脏中合成增加所致,因为给予内毒素后肝脏Vn mRNA的稳态水平增加了4倍。此外,内毒素并未显著增加心脏、肺和大脑中的Vn mRNA水平。这些结果表明,体内Vn基因表达以组织特异性方式受到调控,并将Vn鉴定为一种新型急性期反应物。