Tau蛋白通过外泌体传递到细胞外空间的研究

Study on the Delivery of Tau Protein to Extracellular Space of Cells via Exosomes

目的:验证HEK293-Tau细胞的外泌体中是否存在Tau蛋白、及Tau蛋白是否能够在细胞间通过外泌体传播。方法:通过经典的超速离心结合超滤法分离HEK293-Tau细胞的外泌体,通过透射电镜分析外泌体形态、采用Western blot鉴定外泌体特异性标记物(HSP70、CD63及CD9)及外泌体和细胞培养液中Tau蛋白表达。结果:超速离心结合超滤法成功地分离了HEK293-Tau细胞的外泌体,在透射电镜下观察到外泌体呈现典型的杯状形态,分离的外泌体中检测到HSP70、CD63及CD9等外泌体特异性标记物的表达;在外泌体及细胞培养液中能够检测到Tau蛋白的存在。结论: HEK293-Tau 细胞外泌体中存在Tau蛋白,并且Tau蛋白可以通过外泌体传递到细胞外空间。

Objective: To examine whether Tau protein exists in exosomes isolated from supernatants derived from culture media with HEK293-Tau and whether the Tau protein can be delivered into extracellular space via exosomes. Methods: Exosomes were isolated using ultracentrifugation combined with ultrafiltration.The morphology of isolated exosomes were observed by a transmission electron microscopy.The marker of exosomes was further confirmed using Western blot. Tau protein in isolated exosomes and cell culture medium was also checked using Western blot. Results: Isolated exosomes showed typically cup-shaped morphology.In addition,the exosome marker HSP70,CD63 and CD9 and Tau protein are able to detect in exosomes and cell culture medium. Conclusions: Tau present in cell culture media might be delivered by exosomes derived from HEK293-Tau cells.