摘要
目的:对比4种开窍药有效组分促进替莫唑胺(TMZ)进入人胶质瘤U251细胞内的作用及对细胞增殖、凋亡的影响,并探讨与耐药p-糖蛋白(P-gp)、多耐药基因(MDR1)的关系。方法:体外培养U251细胞,以4种开窍药联合TMZ作用后,高效液相色谱法测定细胞内外TMZ含量,CCK-8法检测细胞增殖,流式细胞术(FCM)检测细胞凋亡,FCM、细胞免疫组化(ICC)/免疫荧光(IF)检测P-gp表达,RT-PCR检测MDR1的表达。结果:4种开窍药均可促进TMZ进入细胞内,TMZ+β-细辛醚组肿瘤细胞增殖显著下降(P<0.05),TMZ+β-细辛醚组、TMZ+冰片组细胞凋亡显著增加,P-gp及MDR1的表达显著降低(P<0.05)。结论:β-细辛醚、冰片、苏合香挥发油、麝香酮均可促进TMZ进入细胞内,增强TMZ抑制肿瘤细胞增殖并促进细胞凋亡,其机制可能是降低P-gp,MDR1的表达。
Objective: To compare the effects of the 4 effective constituents of resuscitation-inducing aromatic herbs inpromoting TMZ into U251 cells and the influence of proliferation and apoptosis of cells. The Counting Kit-8(CCK-8) and apoptosis were tested first, and then we detected the relationship of the effect with P-glycoprotein(P-g) and Multi-drug resistance gene(MDR1). Methods: Then High Performance Liquid Chromatography(HPLC) was used to determinate the TMZ both intracellularly and extracellularly. Morphological change of cells was observed by the microscope. CCK-8 assay was used to measure the cell proliferation-toxicity. Flow Cytometry(FCM) was used to detect the cell apoptosis. Cell immunohistochemistry/immunofluorescence(ICC/IF) and FCM were synchronous used to examine the expression of P-gp. The levels of MDR1 mRNA were also determined by RT-PCR. Results: The 4 effectiveconstituents of resuscitation-inducing aromatic herbs could promote TMZ into U251 cells through membrane, and cell proliferation declined and expression of P-gp/MDR1 decreased while cell apoptosis increased. Conclusion: All the data suggested that co-administration might contribute to the treatment by promoting TMZ into cells and inhibiting P-gp/MDR1 expression.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2017年第5期2206-2209,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
关键词
醒脑开窍药
替莫唑胺
凋亡
P-糖蛋白
多耐药基因
Resuscitation-refreshment herbs
Temozolomide
Apoptosis
P-glycoprotein
Multi-drug resistance gene
