培养人脐静脉内皮细胞慢性缺血缺氧模型

The model of chronic ischemia and hypoxia of cultured human umbilical vein endothelial cells

目的 建立改良的体外培养人脐静脉内皮细胞缺血缺氧的模型。方法 取培养人脐静脉内皮细胞 ,用无菌石蜡油覆盖于低糖细胞培养液表面 ,使培养细胞处于缺血缺氧状态。检测缺血缺氧细胞的形态、培养液中乳酸脱氢酶漏出量及Ⅷ因子相关抗原表达及细胞死亡率。结果 与对照组相比 ,随着缺血缺氧时间的增加 ,内皮细胞形态发生明显改变 ,细胞浆内出现空泡样变性 ,细胞死亡率增加 ,Ⅷ因子相关抗原表达及细胞乳酸脱氢酶漏出量均显著增加 (P <0 .0 1 )。结论 该模型可使细胞受到渐进性损伤 ,损伤结果与体内细胞缺血缺氧后所致的病理学变化相似 ,证实该模型具有一定的可靠性、操作简便、可重复性强。

Objective To establish a modified ischemic and hypoxic model of cultured human umbilical vein endothelial cells(HUVEC) in vitro. Methods HUVEC was cultured in the buffer with low concentration of glucose and covered by sterile paraffin oil. In this model, cultured HUVEC was subjected to insufficient blood and oxygen environment. The cell morphology, leaking lactate dehydrogenase(LDH),cell mortality and Ⅷ relative antigen expression were tested. Results Compared with the control group, the cellular morphology was significantly changed as the increasing time course of cellular ischemia and hypoxia. Vacuolar degeneration occurred in the cell plasm. The cellular mortality, the leaking of LDH, and expression of Ⅷ relative antigen increased significantly( P < 0.01). Conclusions In this chronic ischemic and hypoxic cellular model, cultured cells could be injured gradually, and those cell pathological changes were similar to those in vivo. So our results suggest that the model is reliable and reproducible and simply manipulated.