麝黄消瘤汤对人肝癌细胞多药耐药性逆转作用的实验研究  

Experimental Studies on Reversion Effects of Shehuang Xiaoliu Decoction on Multidrug Resistance of Human Hepatoma Cells

作　　者：张晓丽[1] 黄挺[1] 杨雪飞[1] 黄伶[1] 李勇[1] 

机构地区：[1]浙江省杭州市中医院,杭州310007

出　　处：《中国中医药科技》2014年第1期25-27,32,共4页Chinese Journal of Traditional Medical Science and Technology

摘　　要：目的:通过观察麝黄消瘤汤对细胞内阿霉素浓度、细胞膜转运蛋白(P-gp)表达以及多药耐药基因表达的影响,探究其逆转人肝癌细胞系耐药的分子生物学机制。方法:制备麝黄消瘤汤含药兔血清,配制成15%、20%、25%、30%等不同浓度,培养人肝癌细胞系(Bel-7402),测定麝黄消瘤汤含药兔血清干预前后阿霉素诱导的细胞半数致死量,计算耐药倍数及逆转耐药倍数;应用荧光分光光度计检测细胞内阿霉素浓度,以流式细胞仪检测Pgp表达,以RT-PCR技术检测MDR1mRNA表达。结果:Bel-7402ADR细胞的耐药倍数约为93.5倍。加入20%含药血清后,其IC50降低为9.3μg/ml,由此计算出麝黄消瘤汤含药血清对Bel-7402ADR细胞的增敏倍数为8.3倍;不同浓度的含药血清作用均可降低Bcl-7402细胞的P-gp阳性细胞率,且呈明显的量效关系,与对照组比较有显著意义;麝黄消瘤汤含药血清可抑制人肝癌细胞中MDR基因的表达,且以20%含药血清浓度对MDR的抑制效果最突出,抑制作用随着作用时间的延长而增强,具有时间依赖关系。结论:麝黄消瘤汤能通过提高肿瘤细胞内化疗药物浓度,降低细胞膜转运蛋白的表达,下调多药耐药基因的表达,从而达到逆转人肝癌细胞多药耐药性产生的作用。Objective： To explore the molecular biology mechanism of reversion effect on drug resistance of human hepatoma cells by studying the effects of Shehnang xiaoliu decoction on intracellular adriamycin concentra- tion, expression of transporters protein of cell membrane （Pgp） and expression of multidrug resistance genes. Meflmds： Rabbits ＇ serum containing Shehnang xiaoliu decoction was prepared and divided to different concen- trations of 15%, 20%, 25% and 30%. Human hepatoma cell lines （Bel-7402） were cultured. LDs0 of cells induced by adriamyein was measured both before and after the intervented with serum containing Shehmmg xia- oliu decoction, resistant factor and reversal fold were calculated; fluorospectro photometer was used to detect the concentration of intracellular adriamycin, and flow eytometry for the expression of Pgp, and RT - PCR technique for the expression of MDR1 mRNA. Results： Resistant factor of Bel - 7402ADR was about 93. 5 times. The IC.50 decreased to 9. 3μg/ml after the addition of 20% medicine serum, from which sensitszation of Shehuang xi- aoliu decoction serum to Bel - 7402ADR was calculated to be 8.3 times ; medicine serums of all different concen- tration could decrease rate of P - gp positive cells with an obvious dose - effect relationship and there was signifi- cant difference compared with control group; serum containing Shehnang xiaaliu decoction could inhibit the ex- pression of MDR genes in human hepatoma cells, of which a 20% concentration had the most prominent inhibi- tion effect that enhanced as the processing time prolonged; the inhibition effect enhanced as processing time pro- longed which indicated the time - rely relationship. Conclusions： Shehuang xiaoliu decoction could increase the concentration of intracellular chemotherapy drug of tumors, reduce the expression of transporters protein on cell membrane , and lower the expression of multiclrug resistance genes in order to reverse the influence of mulfidrug resistance of human hepatoma ceUs.

关 键 词：麝黄消瘤汤 人肝癌细胞多药耐药性逆转 基因表达 

分 类 号：R273[医药卫生—中西医结合]