免疫细胞化学法检测肾素(前体)受体在人脐静脉内皮细胞中表达

Expression of(pro)renin receptor in human umbilical vein endothelial cells detected by immunocytochemistry

目的既往研究表明,(P)RR存在于肾系膜细胞、血管平滑肌细胞、心脏、大脑等细胞和器官中,具有升高血压,导致蛋白尿和肾小球硬化等作用。有关(P)RR在内皮细胞表达的报道仍然有限,本研究应用免疫细胞化学法探讨(P)RR在人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)中表达。方法在无菌条件下,取健康产妇分娩后的新鲜婴儿脐带,获得内皮细胞,体外原代培养HUVECs,以免疫细胞化学法鉴定(P)RR在血管内皮细胞中的表达。结果实验获得生长状态良好的内皮细胞。倒置显微镜下可见内皮细胞呈多角形和单层铺路石样排列。经过Ⅷ因子免疫细胞化学染色,棕褐色物质核周围分布密集,细胞边缘颗粒相对较少Ⅷ因子相关抗原阳性,提示该细胞为HUVECs。未加(P)RR抗体的对照组,可见呈紫色的苏木素复染的细胞核,未见细胞膜及细胞浆,免疫细胞化学染色呈阴性。加入(P)RR抗体的(P)RR组,呈DAB染色阳性的棕褐色反应产物存在于细胞膜上和细胞浆中,提示免疫细胞化学(P)RR相关抗原阳性。结论HUVECs中存在(P)RR的表达,可能参与动脉粥样硬化的形成机制。

Objective Previous studies have shown that （pro）renin receptor [（P）RR] exists in the mesangial cells of kidney, vascular smooth muscle cells, heart, and brain. It can increase blood pressure, leading to proteinuria and glomerulosclerosis. There are still few studies on the expression of （P）RR in endothelial cells. This study aims to investigate the expression of （P）RR in the human umbilical vein endothelial cells （HUVECs） by immunocytochemistry. Methods Under sterile conditions, HUVECs were taken from the fresh umbilical cord of infants born to healthy mothers, and primary culture was performed in vitro. The expression of （P）RR in HUVECs was detected by immunocytochemistry. Results Endothelial cells in good growth were obtained. These endothelial cells were in polygonal shapes and arranged like a single layer of paving stones under an inverted microscope. After the factor VIII immunocytochemical staining, brown substance was distributed densely around the nucleus, and there were relatively few particles positive for factor VIII related antigen at the cell edges, suggesting that the cells were HUVECs. In the controlgroup, where （P）RR antibody was not added, the purple nucleus counterstained with hematoxylin was visible, without cell membrane and cytoplasm, and immunocytochemical staining was negative. In the （P）RR group, where （P）RR antibody was added, brown reaction product, which indicated positive DAB staining, was present in the cell membrane and cytoplasm, suggesting that HUVECs were positive for （P）RR related antigen. Conclusion （P）RR is expressed in HUVECs, which may contribute to the development of atherosclerosis.