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目的 :进一步观察amylin对血管内皮细胞屏障功能的影响。 方法 :采用内皮细胞二室弥散系统观察amylin对人脐静脉内皮细胞系 (HUVEC)通透性的影响。通透性用生物素标记的白蛋白跨内皮细胞单层的通透率表示。用囊泡结构特异性探针FM1- 4 3标记内皮细胞囊泡结构 ,在激光共聚焦显微镜下观察并进行定量测定。 结果 :内室单层内皮细胞在amylin浓度为 0、0 1、1,10和 10 0 (nmol/L)的处理 12h后 ,内皮细胞白蛋白通透率分别是 :(5 8± 0 80 )、(6 1± 0 4 2 )、(9 4± 0 35 )、(18 2± 1 72 )和 (2 2 7± 3 9) % ,随amylin浓度的增加白蛋白通透率相应增加。而amylin(10 0nmol/L)与内皮细胞共孵育在 0、0 2 5、0 5、1、3、6、12h白蛋白通透率分别是 :(5 4± 0 3)、(5 8±0 5 )、(6 3± 1 1)、(9 8± 0 6 )、(11 6± 0 3)、(16 9± 1 6 )、(2 0 2± 5 1) % ,随amylin作用时间的延长白蛋白通透率亦相应增加。内皮细胞胞浆内囊泡结构用特异分子探针染色后清晰可见。浓度为 0、0 1、1、10、10 0nmol/L的amylin与内皮细胞共孵育 2h后 ,囊泡吞噬结构染色的平均荧光强度分别是 :2 0 2± 2 6、2 30± 39、36 1± 4 1、4 5 6± 5 7和 72 0±84 /mm2 。随amylin剂量的增加荧光强度相应地增加。amyl
Objective: To further observe the effect of amylin on the barrier function of vascular endothelial cells. Methods: The effect of amylin on the permeability of human umbilical vein endothelial cell line (HUVEC) was observed by endothelial cell two compartment dispersion system. Permeability is expressed as the permeability of biotin-labeled albumin across the monolayer of endothelial cells. The vesicle structure of endothelial cells was labeled with vesicular structure-specific probe FM1-4 3 and observed under confocal laser scanning microscope. Results: The endothelial cell albumin permeability of endothelial monolayer endothelial cells after treated with amylin for 0, 0, 1, 10, and 10 nmol / L for 12 h were (58 ± 0 80) , (6 1 ± 0 4 2), (9 4 ± 0 35), (18 2 ± 1 72) and (2 2 7 ± 3 9)% respectively. The albumin permeability increased with the increase of amylin concentration. However, the permeation rates of amylin (100 nmol / L) incubated with endothelial cells at 0, 0 2, 5, 5, 1, 3, 6 and 12h were (54 ± 0 3) and (6 3 ± 1 1), (9 8 ± 0 6), (11 6 ± 0 3), (16 9 ± 1 6) and (2 0 2 ± 5 1)%, respectively. The prolongation of albumin permeability also increased accordingly. Endothelial cell cytoplasmic vesicle structure with specific molecular probes clearly visible. The average fluorescence intensity of the phagocytic structure of the vesicles after incubating with amylin at concentrations of 0, 0, 1, 1, 10, 10 nmol / L and endothelial cells for 2 h were 202 ± 2 6, 2 30 ± 39, 36 1 ± 4 1,4 5 6 ± 5 7 and 72 0 ± 84 / mm 2. Fluorescence intensity increased with amylin dose increase. amyl