Human endothelial cells produce orosomucoid, an important component of the capillary barrier
Human endothelial cells produce orosomucoid, an important component of the capillary barrier
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DOI：
10.1152/ajpheart.1999.276.2.h530
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发表时间：
1999-02-01
影响因子：
4.8
通讯作者：
Haraldsson, B
中科院分区：
文献类型：
--
作者：
Sörensson, J;Matejka, GL;Haraldsson, B
血清蛋白类粘蛋白（酸性糖蛋白）是维持正常稳态所需的高毛细血管选择性渗透所必需的。目前尚不清楚蛋白质如何执行其行动，但它似乎有助于电荷屏障。此外，最近的研究表明，内皮糖萼是必不可少的电荷屏障。类粘蛋白合成的主要部位是肝脏，但我们想探讨类粘蛋白在内皮细胞中合成的可能性。建立了人皮肤微血管内皮细胞（HMVEC）的原代培养体系。使用人肝细胞作为阳性对照，并从两种细胞类型制备总RNA。逆转录-聚合酶链反应（RT-PCR）进行，并证明orosomucoid表达。RT-PCR后，通过测序确认PCR产物的身份。对来自HMVEC的总RNA进行的RNA酶保护测定证实了RT-PCR的结果，即，类粘蛋白mRNA由内皮细胞表达。免疫沉淀法证实了肝脏和内皮细胞中类粘蛋白的合成。总之，内皮细胞正常产生类粘蛋白，这是必不可少的毛细血管电荷选择性。我们认为，乳清类粘蛋白发挥其作用，通过与其他成分的内皮糖萼。
The serum protein orosomucoid (al-acid glycoprotein) is needed to maintain the high capillary permselectivity required for normal homeostasis. It is not known how the protein executes its action, but it seems to contribute to the charge barrier. Moreover, recent studies suggest that the endothelial glycocalyx is essential for the charge barrier. The main site of orosomucoid synthesis is the liver, but we wanted to explore the possibility that orosomucoid was synthesized in endothelial cells. Primary cultures of human microvascular endothelial cells (HMVEC) from dermal tissue were established. Human liver cells were used as positive controls, and total RNA was prepared from both cell types. Reverse transcription-polymerase chain reaction (RT-PCR) was performed and demonstrated orosomucoid expression. After RT-PCR, the identities of the PCR products were confirmed by sequencing. RNase protection assay performed on total RNA from the HMVEC confirmed the results from the RT-PCR, i.e., orosomucoid mRNA is expressed by endothelial cells. Synthesis of orosomucoid in both liver and endothelial cells was demonstrated by immunoprecipitation. In conclusion, endothelial cells normally produce orosomucoid, which is essential for capillary charge selectivity. We suggest that orosomucoid exerts its effect by interacting with other components of the endothelial glycocalyx.