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Characterization of proteoglycans expressed in brain vascular cells and regulation of their synthesis

脑血管细胞中表达的蛋白多糖的表征及其合成的调节

基本信息

批准号：
14572080
负责人：
KAJI Toshiyuki
金额：
$ 2.24万
依托单位：
Hokuriku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

We investigated the expression, characteristics and regulation of the synthesis of proteoglycans in cultured human brain microvascular endothelial cells and pericytes. The mRNAs coding for proteoglycan core proteins including perlecan, syndecan-1 syndecan-2, syndecan-3, syndecan-4, versican, biglycan and decorin, were determined by reverse transcription-polymerase chain reaction, and the core proteins were detected by Western blot analysis. The experiments indicate that endothelial cells predominantly synthesize perlecan and biglycan whereas pericytes do syndecan-1, biglycan and decorin. Fluorophore-assisted carbohydrate electrophoresis o heparan sulfate revealed that N-sulfation of glucosamine is higher in pericytes than in endothelial cells. These results suggest tha endothelial cells and pericytes in human brain microvascular tissue express different types of proteoglycans whose glycosaminoglycan chains have different composition of disaccharide units that may involved in the regulation of the functions of both endothelial cells an pericytes. Furthermore, we investigated the regulation of proteoglycan synthesis in human brain microvascular endothelial cells am found that vascular endothelial growth factor (VEGF-A_<165>) induce the synthesis of perlecan without influence to the elongation am disaccharide composition. Further studies in detail should be done to clarify the regulation of proteoglycan synthesis in both human brain microvascular endothelial cells and pericytes.

我们研究了培养的人脑微血管内皮细胞和周细胞中蛋白多糖的表达、特性和合成调控。通过逆转录-聚合酶链反应（RT-PCR）检测编码蛋白聚糖核心蛋白（串珠蛋白聚糖、多配体蛋白聚糖-1、多配体蛋白聚糖-2、多配体蛋白聚糖-3、多配体蛋白聚糖-4、多功能蛋白聚糖、双糖链蛋白聚糖和核心蛋白聚糖）的mRNA，并通过Western blot检测核心蛋白的表达。实验表明，内皮细胞主要合成串珠蛋白聚糖和双糖蛋白聚糖，而周细胞合成多配体蛋白聚糖-1、双糖蛋白聚糖和核心蛋白聚糖。硫酸乙酰肝素的荧光团辅助糖电泳显示，氨基葡萄糖的N-硫酸化在周细胞中比在内皮细胞中更高。这些结果表明，人脑微血管组织中的内皮细胞和周细胞表达不同类型的蛋白聚糖，其糖胺聚糖链具有不同的二糖单元组成，可能参与内皮细胞和周细胞功能的调节。进一步研究了人脑微血管内皮细胞蛋白多糖合成的调控，发现血管内皮生长因子（VEGF-A<165>）诱导串珠素合成，而不影响串珠素的延伸和二糖组成。对人脑微血管内皮细胞和周细胞中蛋白多糖合成的调控还需进一步研究。