[关键词]
[摘要]
目的 运用小鼠脑微血管内皮细胞(bEnd.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bEnd.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50 μmol/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。
[Key word]
[Abstract]
Objective To investigate the effect of borneol on puerarin through in vitro blood-brain barrier (BBB) model and to discuss the main pathway of borneol to promote the opening of BBB. Methods MTT assay was conducted to investigate the toxic effects of borneol and puerarin with different concentration on the cells and to screen the concentration of tested drug. In vitro model of BBB was used to observe the effect of borneol on the opening of tight junction and the effect of borneol on puerarin through BBB. Results The experimental drug concentration of borneol and puerarin was both 50 μmol/L by MTT experiment. There was no significant change in transepithelial electrical resistance (TEER) before administration and 24 h after administration, and the permeation rate of puerarin group and borneol + puerarin group were (59.96 ±5.90)% and (106.80 ±2.73)%, respectively, with significant difference between two groups. Conclusion Borneol combined with puerarin can promote its permeation rate to a certain extent, but its mechanism needs to be further explored by cell-related tight junction proteins level and adenosine receptor signaling pathway.
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[基金项目]
国家自然科学基金资助项目(81573718)