12外泌体对体外血脑屏障模型功能的影响

Effects of Exosomes on the Function of Blood-brain Barrier Model in Mice

目的 在体外建立血脑屏障模型,研究非小细胞肺癌a549细胞来源的外泌体对血脑屏障模型功能的影响。方法 利用小鼠脑微血管内皮细胞与星型胶质细胞建立体外血脑屏障模型。模型建立后通过4 h渗漏实验鉴定是否成模。建模成功后分为对照组与实验组,将利用超速离心法及Exo-spin试剂盒分离提纯的非小细胞肺癌A549细胞培养基中外泌体接种至实验组Transwell上室,置于培养箱孵育,分别于共培养24 h、48 h、72 h后进行跨内皮电阻检测实验及48 h辣根过氧化物酶渗透试验,以评估体外血脑屏障模型的功能。结果(1)80%血脑屏障模型在建立建模成功;(2)2组TEER值比较24 h后2组间差异无统计学意义(P > 0.05);48 h后实验组TEER降低,与对照组比较差异有统计学意义(P < 0.01);72 h后实验组TEER进一步降低且与对照组相比差异有统计学意义(P < 0.01);(3)共培养48 h后实验组通透性较对照组明显升高,差异有统计学意义(P < 0.05)。结论 人非小细胞肺癌细胞外泌体可以影响小鼠体外血脑屏障模型的功能,人非小细胞肺癌细胞外泌体可使BBBM通透性增加。

Objective To study the effects of exosome derived from non-small cell lung cancer A549 cells on the function of blood-brain barrier model by establishing a blood-brain barrier model in vitro. Methods An in vitro blood-brain barrier model was established using mouse brain microvascular endothelial cells and astrocytes. After the model was established, it was identified by a 4h leak test. After successful modeling, the patients were divided into the control group and the experimental group. The exosomes in the non-small cell lung cancer A549 cell culture medium purified by ultracentrifugation and Exo-spin kit were inoculated into the transwell upper chamber of the experimental group and placed in the incubator. Trans-endothelial resistance test and 48h horseradish peroxidase permeation test were performed after co-culture for 24h, 48h and 72h respectively to evaluate the function of the blood-brain barrier model in vitro. Results (1)80% blood-brain barrier models were successfully modeled.(2)The comparison of TEER values between the two groups: there were no significant changes between the two groups after 24h (P > 0.05);after 48h, the TEER of the experimental group decreased compared with the control group, which was statistically different from the control group (P < 0.01);after 72h, the experimental group TEER further decreased,which was statistically different from the control group (P <0.01).(3) The permeability of the experimental group was significantly higher than that of the control group after 48 hours of co-culture, and there was statistical difference (P < 0.05). Conclusion Small cell lung cancer cell exosomes can affect the function of the mouse blood-brain barrier model in vitro. Human non-small cell lung cancer cell exosomes can increase BBBM permeability.