脑微血管内皮细胞氧糖剥夺条件培养液对PC12细胞活性的影响

Effects of Conditioned Medium of Microvascular Endothelial Cells of Rat Brain after the Deprivation of Oxygen and Glucose on PC 12 Cells

目的:观察脑微血管内皮细胞氧糖剥夺后条件培养液对PC12细胞的影响。方法:原代培养大鼠脑皮质微血管内皮细胞,传至三代,制备其正常、氧糖剥夺、复糖复氧三种状态条件液,并观察内皮细胞在这三种状态下的形态改变;将这三种条件培养液作用于PC12细胞,分4组:正常对照组(Normal)、正常内皮细胞条件液组(N-CM)、氧糖剥夺内皮细胞条件液组(I-CM)、复糖复氧内皮细胞条件液组(R-CM),每组分别设6 h、12 h、24 h、48 h、72 h、96 h、120 h、144 h 8个时间点,MTS/PMS法检测三种脑微血管内皮细胞条件液作用后的不同时间点PC12细胞活性的变化。结果:氧糖剥夺后脑微血管内皮细胞发生核固缩等明显病理改变,复糖复氧后这些改变有所恢复。N-CM组、R-CM组与相应时间点的Normal组PC12细胞活性相比差异均有统计学意义(P<0.01)。正常和复糖复氧内皮细胞条件液显著抑制了PC12细胞的增殖和活性。N-CM组、R-CM组与相应时间点的I-CM组PC12细胞活性相比差异均有统计学意义(P<0.01)。结论:正常脑微血管内皮细胞条件液抑制了PC12细胞的活性,氧糖剥夺后的脑微血管内皮细胞条件液抑制效应消失,复糖复氧后,这种抑制效应也同时恢复。

Objective：To observe the effects of conditioned medium of microvascular endothelial cells of rat brain after the deprivation of oxygen and glucose on PC 12 Cells.Methods：The primary cultured microvascular endothelial cells of rat brain were passed into 3rd passage and three kinds of conditioned medium was collected：normal,deprivation of oxygen and glucose and restoration of oxygen and glucose.Morphology of microvascular endothelial cells was compared.Then the three conditioned medium added into the PC12 cells and separated them into 4 groups：Normal,N-CM,I-CM and R-CM.Each group was separated into 6h,12h,24h,48h,72h,96h, 120h and 144h.The activities of PC 12 cells was determined through the measurement of MTS/PMS.Results：Karyopcnosis was observed in the microvascular endothelial cells of the deprivation of oxygen and the little reablement was found after restoration of oxygen and glucose.N-CM and R-CM groups showed significant inhibition effect on the activity of PC12 cells compared to Normal groups（P〈0.01）. N-CM and R-CM groups showed significant inhibition effect on the activity of PC 12 cells compared to I-CM groups（P〈0.01）.Conclu- sion：The conditioned medium of normal microvascular endothelial cells of rat brain could inhibit the activity of PC cells,the inhibition was disappeared in the conditioned medium of deprivation of oxygen and glucose,as well as the inhibition was returned after restoration of oxygen and glucose.