猪血脑屏障内皮细胞微囊的制备与鉴定

Preparation and Identification of Micro-vesicles from Endothelial Membrane of Blood Brain Barrier of Sows

选二花脸母猪和长白母猪各 5头 ,取大脑皮质 ,制备微血管 ,分离内皮细胞微囊 ,应用倒置显微镜和透射电镜观察微囊的完整性 ;通过测定γ-谷氨酰转肽酶、5′-核苷酸酶、乙酰胆碱酯酶、乳酸脱氢酶、碱性磷酸酶等标志酶活性 ,鉴定微囊的纯度和生物学活性。结果表明 ,用该方法分离得到的大脑皮质内皮细胞微囊的完整性较好 ,纯度较高 ,生物学活性好 。

The micro-vesicles were isolated and purified from the endothelial membrane of micro-vessels on cerebral cortex of 5 for each Erhualian and large white sows. The integrity of the vesicles was evaluated by inverted microscopy and electron microscopy. The purity and biological activity were determined by the activities of such enzymes as g-glutamy transpeptidase (g-GTase), 5′-nucleotidase (NTDase), acetylcholinesterase(ACSase), lactate dehydrogenase(LDHase) and alkaline phosphatase (AKPase). The results indicated that thd endothelial membrane vesicles isolated by the above-mentioned method is of high purity and biological activity and may be used to evaluate the transport via blood-brain barrier.