[关键词]
[摘要]
本文运用Western blot技术来研究右旋龙脑对细胞膜上ABC转运蛋白ABCB1、ABCC1和ABCG2表达量的影响,并进一步运用流式细胞仪考察右旋龙脑对肝癌细胞HepG2细胞膜通透性的影响,探讨其可能作用的物理模型。结果表明:右旋龙脑能够显著下调转运蛋白ABCB1、ABCC1和ABCG2的表达量,增加HepG2细胞的细胞膜通透性,从而提高姜黄素类化合物在HepG2细胞内的含量。当用20 μg/mL NB前处理HepG2细胞12 h后再用20 μM Cur、40 μM DCur和40 μM BDCur分别处理24 h,细胞内ABCB1的蛋白表达量从0.7、0.9和0.7降低到0.4、0.6和0.5。当用20 μg/mL NB前处理HepG2细胞12 h后再用40 μM BDCur处理24 h,细胞内ABCC1和ABCG2的蛋白表达量从1.0和0.7分别降低到0.5和0.3。进一步的流式结果表明,右旋龙脑能够提高 HepG2细胞的细胞膜通透性,当用20 μg/mL NB处理HepG2细胞12h后,细胞内YO-PRO-1的荧光强度从对照组的20.79升高到45.51,同时其荧光强度与NB呈浓度依赖性。
[Key word]
[Abstract]
Western blotting was used to study the impact of d-borneol on the expression of ABC transporter proteins ABCB1, ABCC1, and ABCG2 in the cell membrane. Furthermore, flow cytometry was used to examine the impact of d-borneol on the membrane permeability of HepG2 cells, and a possible physical model for the action of d-borneol was investigated. The results showed that d-borneol significantly downregulated the expression levels of the transport proteins ABCB1, ABCC1, and ABCG2, and increased HepG2 cell membrane permeability, thus improving the absorption of curcuminoids. When HepG2 cells were pretreated with 20 μg/mL d-borneol for 12 h, followed by 24-h treatment using 20 μM curcumin (Cur), 40 μM demethoxycurcumin (DCur), and 40 μM bisdemethoxycurcumin (BDCur), respectively, the expression levels of ABCB1 in cells reduced from 0.7, 0.9, and 0.7 to 0.4, 0.6, and 0.5, respectively. When HepG2 cells were pretreated with 20 μg/mL d-borneol for 12 h, followed by 24-h treatment using 40 μM BDCur, the expression levels of ABCC1 and ABCG2 in the cells were reduced from 1.0 to 0.7 and from 0.5 to 0.3, respectively. Further flow cytometric analysis indicated that d-borneol could improve the membrane permeability of the HepG2 cell. When HepG2 cells were pretreated with 20 μg/mL d-borneol for 12 h, the fluorescence intensity of YO-PRO-1 in the cells increased from 20.79 (control group) to 45.51, and the fluorescence intensity was positively correlated with the concentration of d-borneol.
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[基金项目]
广东省自然科学基金面上项目(2014A030313265);广东省扬帆计划项目(201312H05);广东省科技计划项目(2015A020209020,2015A020209036);华南理工大学中央高校基本科研业务费资助。