一种单核细胞渗出模型的建立及其在内皮细胞通透性检测中的应用

Establishment of one kind of monocyte diapedesis model and its application in assessment of endothelial cells permeability

目的建立一种单核细胞渗出模型并探讨其作为内皮细胞通透性检测模型的可行性。方法将培养的人脐静脉内皮细胞与浓度为100μg／ml晚期糖基化终产物修饰的人血清白蛋白分别共同培养2、4、6、8、12h，同样浓度的人血清白蛋白加入人脐静脉内皮细胞作为对照组，然后采用Transewell模型，利用5一氯甲基二乙酸荧光素标记的人白血病单核细胞渗出数量反映内皮细胞的通透性。结果与对照组相比，晚期糖基化终产物修饰的人血清白蛋白以时间依赖的方式增加白血病单核细胞渗出的数量，且在刺激8h组白血病单核细胞渗出的数量达高峰（P〈0．05）。结论用5一氯甲基二乙酸荧光素标记的白血病单核细胞建立的单核细胞渗出模型为体外研究内皮细胞通透性提供了一种简便、可行的方法。

Objective To establish one kind of monocyte diapedesis model and study its application in the assessment of endothelial cells permeability. Methods Human umbilical vein endothelial eells（HUVECs） were cultured in vitro with AGEs-modified human serum albumin（AGEs-HSA） in the concentration of 100 μg/ml for 2,4,6,8 and 12 hours respectively. In control group, HSA of the same concentration was administered to HUVECs cells. By using transewell model, the permeability of endothelial cells was presented by the amount of permeated 5-chloromethylfluorescein diacetate（CMF- DA）-labeled human acute monocytic leukemia cell line（THP-1） cells. Results Compared to controls, AGEs-HSA increased the amount of THP-1 cells in a time-dependent manner, and the amount of permeated THP-I cells reached the peak at 8 hours in stimulating group（P〈0. 05）. Conclusions The monocyte diapedesis model based on CMFDA-labeled human THP-1 monocytes provides a simple and reliable means for studying the permeability of endothelial cells in vitro.