川芎嗪与β-榄香烯联合应用诱导K562/ADM细胞凋亡及逆转其MDR的实验研究

The Studies of Tetramethylpyrazine in Combination with β- Elemene on Induction of the Apoptosis and the Reversal of Multidrug Resistance in K562/ADM Cells Line

目的:探讨不同作用机制的川芎嗪(Tetramethylpyrazine,TMP)与β-榄香烯(β-elemene)联合应用,从多环节逆转耐药细胞K562/ADM的多药耐药(multidrugresistance,MDR),以期进一步提高逆转效果。方法:采用MTT法检测细胞的药敏性及抗药性逆转,流式细胞术及透射电镜检测细胞凋亡,应用SPSS(10.0ProductionPacility)软件包对实验结果进行统计学处理。结果:1)非细胞毒性浓度的TMP(350μg/ml)及β-榄香烯(4.0μg/ml)可显著降低ADM对K562/ADM细胞的IC50(P<0.01),逆转耐药倍数分别为2.03倍及2.18倍。β-榄香烯(4.0μg/ml)具有诱导该细胞凋亡的作用。2)上述两种药物以非细胞毒性浓度联合应用,对ADM的逆转倍数为4.65倍,明显高于二者单独应用,而且也高于两者单独应用之和。其主要逆转机制是两药的联合应用通过升高细胞内ADM的浓度,诱导该细胞凋亡的途径,实现提高逆转效果的目的。结论:TMP和β-e榄香烯联合应用能明显逆转K562/ADM细胞对ADM的耐药性和诱导该细胞的凋亡。

Objective: To further improve the reversed effect, we attempted to take the Tetramethylpyrazine (TMP) in combination with β-elemene which were different in effect mechanism, and reversed the MDR (multidrug resistnce) of K562/ADM via more links. Methods: The sensitivity of cell lines and the reversal of resistance were determined with MTT method. The cell apoptosis was determined by flow cytometry and transmission electron microscope. Statistical treatment with SPSS was applied to deal with all the experimental data. Results: 1, The non-cytotoxic dose of TMP (350μg/ml)and β-elemene (4.0μg/ml) could markedly decrease the IC50 value of K562/ADM cell line to adriamycin(ADM) and increase the sensitivity of K562/ADM cell line to ADM. The resistance-reversing effect was 2.03-fold and 2.18-fold respectively. The non-cytotoxic dose of β-elemene (4.0μg/ml) could induce apoptosis effectively; 2, The non-cytotoxic dose of TMP combined with β-elemene strongly improved drug resistance reversal effect (4.65-fold), clearly, which was larger than the single application of TMP(350μg/ml) or β-elemene(4.0μg/ml) respectively and their summation. We also found these two medicines were helpful to increase the intracellular ADM concentration and to induce cell apoptosis. Conclusion: Administration of TMP in combination with β-elemene has obvious co-effect on the resistance-reversing and the inducement of apoptosis in K562/ADM cell line.