培养的大鼠脑微血管内皮细胞生化特性观察

OBSERVATION OF THE BIOCHEMICAL CHARACTERS OF CULTURED MICROVASCULAR ENDOTHELIAL CELLS FROM RAT BRAIN

用胶原酶消化、差异离心和尼龙网过滤的方法分离鼠脑微血管内皮细胞 ,建立其体外长期培养方法。经形态学 ,免疫组化 ,酶学等鉴定 ,培养细胞为脑微血管内皮细胞。动态观察培养细胞酶含量变化 ,发现随着细胞培养时间的延长 ,血管紧张素转换酶 (ACE)呈上升趋势 ,而γ-谷氨酰胺转化酶 (γ- GT)和硷性磷酸酶 (AL P)则明显下降。实验结果提示 ,血脑屏障的主要功能酶γ- GT和 AL P可作为体外脑微血管内皮细胞的标志酶 ,但要维持长时间体外表达则需要某种因子的介导。本实验可为体外研究血脑屏障及相关疾病提供帮助。

Brain microvascular endothelial cells (BMEC) of Spnaque Dawley weanling rats were prepared by collagenase digestion, differential centrifugation and filtering through a nylon mesh. Cells were cultured in vitro for long term. Using morphological, immunohistochemical and enzymatic methods, the cultivated cells were affirmed as microvascular endothelial cell of brain. Observing the changes of enzymatic activity, we found that the longer the BMEC cultured in vitro, the higher activity the ACE was. On the contrary, the activity of γ GT and ALP were lower. This result might suggest that γ GT and ALP, which are the most important functional enzymes of blood brain barrier, could serve as an enzymatic marker for cultured BMEC. However, an induced factor was needed for maintaining cells producing and secreting these enzymes for a long time. In addition, this paper provided a useful method for studying the blood brain barrier in vitro.