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目的探讨纳米囊制剂对白血病多药耐药细胞的影响。方法采用溶剂挥发法制备共载中药甘草酸(GA)及化疗药物柔红霉素(DNR)的纳米囊,将其分为P-gp Ab修饰共载中药GA及化疗药物DNR组(A组);P-gp Ab修饰装载DNR化疗药物组(B组);DNR化疗药物组(C组);DNR+GA组(D组);P-gp Ab修饰纳米囊空载组(E组);GA组(F组);生理盐水对照组(G组)。采用体外四甲基偶氮唑盐(MTT)比色法评价细胞毒作用(白血病多药耐药细胞K562/A02),采用流式细胞仪检测细胞凋亡的情况,采用化学发光自显影免疫印迹法(Western Blot)检测蛋白B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)的表达。结果 A组的K562/A02的半抑制浓度指数(IC_(50))为(3.65±1.06),其降低效果优于其他组(P<0.05)。B组IC_(50)为(3.12±0.96),低于C、D、F组的(0.95±0.04)、(0.96±0.03)、(0.94±0.05)(P<0.05)。C、D、F组的耐药指数(RE)均接近1,故认为对K562/A02的细胞无增强耐药作用,也无毒性作用。E组和G组为对照组,检测到的细胞掉凋亡率较小,不计入比较。A组的细胞总凋亡率高于其他各组(P<0.05)。B组的细胞凋亡率高于C、D、F组(P<0.05)。A组的Bcl-2的表达低于其他各组,Bax的表达高于其他各组(P<0.05)。E组和G组的Bcl-2表达均高于其他各组,Bax的表达均低于其他各组,差异均具有统计学意义(P<0.05)。结论纳米囊制剂对白血病多药耐药细胞具有细胞毒性,中药GA+化疗药物DNR具有明显的协同效果,其作用机制可能与上调Bax表达与下调Bcl-2有关。
Objective To investigate the effects of nanocapsules on multidrug-resistant leukemia cells. Methods Nanocapsules loaded with glycyrrhizin (GA) and chemotherapy drug daunorubicin (DNR) were prepared by solvent evaporation method, and were divided into three groups: group A (GA) and chemotherapy group DNR (Group B); DNR chemotherapy group (group C); DNR + GA group (group D); P-gp Ab modified nanocapsules no-load group Group (Group F); saline control group (Group G). Cytotoxicity (K562 / A02, a multidrug-resistant leukemia cell line) was evaluated by MTT colorimetric assay. The apoptosis of the cells was detected by flow cytometry. Western Blot was used to detect the expression of Bax and Bcl-2. Results The half inhibitory concentration index (IC 50) of K562 / A02 in group A was (3.65 ± 1.06), which was significantly lower than that in other groups (P <0.05). The IC 50 in group B was (3.12 ± 0.96) lower than that in group C, D and F (0.95 ± 0.04), (0.96 ± 0.03) and (0.94 ± 0.05), respectively (P <0.05). The drug resistance index (RE) of C, D and F groups were close to 1, so there was no enhancement of resistance to K562 / A02 cells and no toxic effect. E group and G group as the control group, the detected cell apoptosis rate is small, not included in the comparison. The total apoptosis rate of group A was higher than that of other groups (P <0.05). The apoptosis rate of group B was higher than that of group C, D and F (P <0.05). The expression of Bcl-2 in group A was lower than that in other groups, and the expression of Bax in group A was higher than that in other groups (P <0.05). The expression of Bcl-2 in E group and G group were higher than those in other groups, and the expression of Bax was lower than other groups (P <0.05). Conclusion The nanocapsule preparation is cytotoxic to multidrug-resistant cells of leukemia. The synergistic effect of GA + chemotherapeutic drug DNR is obvious. Its mechanism may be related to up-regulation of Bax expression and down-regulation of Bcl-2.