蟾酥注射液逆转HL60/阿霉素细胞多药耐药的机制

Mechanisms of Chansu Injection in reversing multidrug resistance of HL60/ADM cells

目的探讨蟾酥注射液(CHS)对人类白细胞多药耐药细胞系HL60/阿霉素细胞的逆转作用机制。方法 MTT法测定CHS预作用后HL60/阿霉素细胞对阿霉素的敏感性;流式细胞术考察CHS对HL60/阿霉素细胞周期的影响;免疫组化考察CHS对HL60/阿霉素细胞NF-κB、MRP、GST-π、iNOS蛋白表达的调节作用。结果 CHS可将阿霉素对HL60/阿霉素细胞的IC50值由34.1971μmol/L降至17.4393μmol/L,逆转倍数为1.9609倍;FCM显示CHS作用后HL60/阿霉素阻滞于G0/G1期、G2/M期,降低进入S期比例,并出现凋亡峰;免疫组化结果显示,CHS能下调HL60/阿霉素细胞中MRP、GST-π表达,上调NF-κB、iNOS的表达。结论蟾蜍注射液逆转HL60/阿霉素细胞的多药耐药性可能是通过下调MRP、GST-π的表达,上调NF-κB、iNOS的表达,促进HL60/阿霉素细胞凋亡,从而增加HL60/阿霉素细胞对药物的敏感性。

Objective To investigate the mechanisms underlying the effect of Chansu Injection（CHS） in reversing multi-drug resistance（MDR） of HL60/ADM cells.Methods MTT assay was used to investigate the effect of CHS on adramycin（ADM） sensitivity of HL-60/ADM cells.Flow cytometry was used to observe the effect of CHS on the cell cycle of HL60/ADM cell.The expressions of NF-κB,MRP,GST-π,and iNOS were detected by immunocytochemistry.Results Treatment with CHS lowered the IC50 of ADM in HL60/ADM cells from 34.1971 μmol/L to 17.4393 μmol/L,and caused an increase in G0/G1 and G2/M phase cells with decreased S phase cells.CHS decreased the expressions of MRP mRNA and GST-π and MRP proteins but increased the expressions of iNOS and NF-κB proteins in the cells.Conclusion CHS can partly reverse MDR in HL60/ADM cells possibly by down-regulating MRP and GST-π,up-regulating NF-κB and iNOS,and promoting cell apoptosis,thereby increase ADM sensitivity of HL-60/ADM cells.