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目的构建并观测血脑/血瘤屏障体外模型的形态与功能特性,为中枢神经系统药物跨血脑和(或)血瘤屏障研究提供体外实验模型。方法将分离的Balb/C小鼠脑微血管内皮细胞 (BMEC)在铺有明胶的微孔膜上单层培养或与大鼠胶质瘤细胞C6双室共培养,分别建立血脑屏障 (BBB)和血瘤屏障(BTB)模型,采用光镜和扫描电镜观察BMEC形态,采用Millicell-ERS系统检测屏障中的跨内皮电阻(TEERs),免疫细胞化学检测P-糖蛋白(P-gp)表达,并比较BBB和BTB中 BMEC的形态和功能特征。结果两组模型中的BMEC均形成单层生长和良好的细胞间紧密连接, 产生较高的TEERs值,并检测到P-gp表达。瘤细胞诱导使BMEC间出现间隙,同时相点TEERs值降低,P-gp表达减弱。结论两种体外模型可用于研究药物跨血脑和(或)血瘤屏障特性,其中BTB 模型可能更适合抗肿瘤药物的研究。
Objective To construct and observe the morphological and functional characteristics of the in vitro model of the blood-brain / blood-brain barrier and to provide an in vitro experimental model for the study of CNS drugs across the blood brain and / or the blood-borne barrier. Methods The isolated Balb / C mouse brain microvascular endothelial cells (BMECs) were cultured in monolayers on gelatin-coated microporous membranes or co-cultured with rat C6 glioma cells to establish the blood-brain barrier (BBB) (BTB). The morphology of BMECs was observed under light microscope and scanning electron microscope. The trans-endothelial resistance (TEERs) in the barrier was detected by Millicell-ERS system. The expression of P-glycoprotein (P-gp) was detected by immunocytochemistry. The morphology and functional characteristics of BMEC in BBB and BTB were compared. Results BMEC in both groups formed monolayer growth and good intercellular tight junction, resulting in higher TEERs and P-gp expression. Tumor cells induced a gap between BMECs, while the phase point TEERs decreased, P-gp expression weakened. Conclusions Two in vitro models can be used to study the transmembrane and / or blood barrier properties of the drug. The BTB model may be more suitable for antineoplastic agents.