Mast cell granule heparin proteoglycan induces lacunae in confluent endothelial cell monolayers.
Mast cell granule heparin proteoglycan induces lacunae in confluent endothelial cell monolayers.
复制标题
肥大细胞颗粒肝素蛋白多糖在汇合的内皮细胞单层中诱导腔隙。
DOI：
10.1016/s0002-9440(10)65412-0
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Rickard,A
中科院分区：
文献类型：
--
作者：
Lagunoff,D;Rickard,A
此外，大鼠肥大细胞颗粒汇合牛肺动脉内皮细胞单层导致在培养物中形成许多腔隙。几条证据表明肝素蛋白聚糖是导致该效应的颗粒基质组分：- 颗粒提取物层析后蛋白聚糖级分中存在活性，用肝素酶I消化抑制颗粒活性，用蛋白酶K蛋白水解蛋白聚糖级分不能显著降低其活性，以及糜酶和羧肽酶抑制剂不能抑制颗粒活性。孔形成的发作被推迟了几个小时后，颗粒添加到培养物中，最大的孔形成发生在8和16小时之间，并持续长达24小时。腔隙由单层内活动内皮细胞分离形成，不归因于细胞收缩活性或细胞丢失。延时视频记录显示，这些洞是动态的，单个洞在几个小时内扩大和缩小。明胶和纤连蛋白表面均出现腔隙。颗粒中活性糜酶的存在阻止了蛋白多糖的作用。肝素糖胺聚糖与蛋白聚糖不同，对内皮细胞单层没有类似的影响，但确实阻断了随后加入的颗粒的作用，表明肝素反应性受体或结合位点的可能性。
The addition of rat mast cell granules to confluent bovine pulmonary artery endothelial cell monolayers resulted in the formation of numerous lacunae in the cultures. Several lines of evidence identified heparin proteoglycan as the component of the granule matrix responsible for the effect: presence of the activity in the proteoglycan fraction after chromatography of granule extracts, inhibition of granule activity by digestion with heparinase I, the failure of proteolysis of the proteoglycan fraction with proteinase K to significantly diminish its activity, and the failure of chymase and carboxypeptidase inhibitors to inhibit granule activity. The onset of hole formation was delayed for several hours after granule addition to the culture, and maximal hole formation occurred between 8 and 16 hours and was sustained as long as 24 hours. The lacunae formed by the separation of motile endothelial cells within the monolayer and was not attributable to cell contractile activity or cell loss. Time-lapse video recording showed that the holes were dynamic, individual holes expanding and regressing over a period of hours. Formation of lacunae occurred on gelatin and fibronectin surfaces alike. The presence of active chymase in the granules prevented the action of the proteoglycan. Heparin glycosaminoglycan as distinct from the proteoglycan did not similarly affect the endothelial monolayers but did block the action of granules added subsequently, indicating the likelihood of a heparin-reactive receptor or binding site.