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目的运用小鼠脑微血管内皮细胞(b End.3)制备血脑屏障(BBB)模型,探讨冰片对葛根素透过BBB的影响并初步探讨冰片促进BBB开放的主要途径。方法 MTT实验考察不同浓度冰片和葛根素对b End.3细胞的毒性作用,筛选实验药物浓度。应用BBB体外模型,以跨内皮细胞间电阻作为紧密连接程度的主要反应指标,观察冰片对其紧密连接的开放有无直接影响以及冰片对葛根素跨BBB转运的影响。结果经MTT实验确定冰片和葛根素的实验药物浓度均为50μmol/L。各组给药前与给药24 h后跨膜电阻(TEER)未见明显改变,葛根素组、冰片+葛根素组透过率分别为(59.96±5.90)%和(106.80±2.73)%,差异显著(P<0.05)。结论冰片联合葛根素用药可以一定程度上促进葛根素透过BBB,但其开窍机制还需通过细胞的相关紧密连接蛋白水平和腺苷受体信号通路进一步探讨。
OBJECTIVE: To study the effect of borneol on the BBB permeability of BBB by using mouse brain microvascular endothelial cells (bEnd.3), and to explore the main ways of borneol promoting BBB opening. Methods The toxicity of borneol and puerarin on bEnd.3 cells was investigated by MTT assay and the concentration of experimental drugs was screened. BBB in vitro model was used to determine the effect of borneol on the transport of puerarin across BBB by using the intercellular resistance as the main reaction index of tight junctions. Results The experimental drug concentrations of borneol and puerarin were all determined to be 50 μmol / L by MTT assay. The transmembrane resistance (TEER) did not change significantly in each group before and 24 h after administration. The transdermal permeation rates of puerarin and borneol + puerarin groups were (59.96 ± 5.90)% and (106.80 ± 2.73)% respectively, The difference was significant (P <0.05). Conclusion Borneol combined with puerarin can promote puerarin to penetrate the BBB to a certain degree, but its mechanism of resuscitation needs to be further explored through the expression of cell-related tight junction protein and adenosine receptor signaling.