醋柴胡及其不同部位对BRL3A细胞摄取顺铂及Oct2,Mrp2基因表达的影响

The effect of vinegar-baked Radix Bupleuri and its extracts on the uptake of cisplatin and mRNA expression of Oct2 and Mrp2 in BRL3A cell

目的考察醋柴胡及其不同提取部位对大鼠肝细胞摄取顺铂的影响,并初步探讨其机制,为醋柴胡肝靶向增强作用机制探讨提供数据支撑。方法选用顺铂作为Oct2和Mrp2的共转运底物。对数增殖期内的BRL3A细胞经醋柴胡水提液及其不同部位成分作用24 h后;继续加入顺铂孵育4 h,采用HPLC法测定细胞内顺铂含量,并按蛋白质浓度归一化计算摄取量;另采用实时定量PCR法测定醋柴胡药物作用24 h后胞内Oct2和Mrp2基因的表达量。结果醋柴胡水提取液及正丁醇萃取部位可显著促进顺铂的摄取,醋柴胡多糖部位及小分子水溶性部位则显著降低顺铂的摄取;醋柴胡水提取液显著促进Oct2和Mrp2的基因表达,其它部位则均显著促进Mrp2基因表达,但对Oct2基因表达影响不大。结论醋柴胡肝靶向增强作用与其增加Oct2 mRNA表达有关,正丁醇部位可能为其有效部位;而促进Mrp2基因表达可能与醋柴胡疏肝解郁作用有关。

Objective To study the effect of vinegar-baked Radix Bupleuri（ VBRB） and its extracts on the uptake of cisplatin,and investigate the mechanism by determine the mRNA expression of Oct2 and Mrp2,to shed some light on the meridian guide effect of VBRB. Methods Cisplatin was used as co-substrate of Oct2 and Mrp2. After treated with VBRB and its extracts for24 h,cytoplasmic substrate contents were detected by HPLC method and the uptake rate were normalized by protein concentration,and the mRNA level of Oct2 and Mrp2 were detected by real-time quantitative PCR（ RT-PCR）. Results VBRB（ 10 mg·ml^-1） and n-Butanol fraction（ Buf,50 mg·ml^-1） increased the magnitude of the cellular uptake rate of cisplatin by： 123. 8%,146. 6%（ P〈0. 01）,while polysaccharides（ PSS,10 mg·ml^-1） and micromolecular hydrophilic extract（ MHE,50 mg·ml^-1）reduced the uptake of cisplatin by： 44. 6%（ P〈0. 05） and 56. 1%（ P〈0. 01）. And VBRB（ 10 mg·ml^-1） increased Oct2 mRNA expression by 98. 4%（ P〈0. 05）,Buf,PSS and MHE had marginal effect on the mRNA expression of Oct2. VBRB,SSs,PSS and MHE increased the Mrp2 mRNA expression by 184. 1%,169. 1%（ P〈0. 01）,94. 4% and 81. 9%（ P〈0. 05） respectively. Conclusion VBRB increased liver-targeting may be via increasing the expression of Oct2,and the up-regulation of Mrp2 mRNA may be associated with the effect of dispersing stagnated liver qi for relieving qi stagnation.