槲皮素增敏阿霉素治疗难治耐药急性白血病疗效的研究

Efficacy of Quercetin-sensitized Adriamycin for Treatment of Refractory Acute Leukemia

目的:探讨槲皮素增敏阿霉素的化疗效果。方法:应用CCK-8检测不同剂量阿霉素、槲皮素及槲皮素联合阿霉素对临床难治耐药急性白血病患者的原代白血病细胞增殖的抑制作用;用槲皮素、阿霉素及二者联合用药作用于非照射T-ALL白血病小鼠,观察小鼠生存期的变化,以及对于心肌损伤的程度。结果:在3个不同时间点24、48和72h,不同阿霉素浓度组(6、0.6和0.06μg/ml)与阿霉素半剂量组(3、0.3和0.03μg/ml)+槲皮素浓度0.25mmol/L组两两比较发现,对原代白血病细胞增殖抑制率总体差异无统计学意义;各药物浓度组中原代白血病细胞增殖抑制率组间比较总体差异有统计学意义,对原代白血病细胞增殖抑制作用呈浓度和时间依赖性(r24h,a\c\e=0.995、r48h,a\c\e=1.000、r72h,a\c\e=0.984、r24h,b\d\f=0.993、r48h,b\d\f=0.999、r72h,b\d\f=0.960)。体内实验显示,经低剂量阿霉素联合槲皮素作用的非照射T-ALL白血病小鼠生存期无明显延长,高剂量阿霉素联合槲皮素作用的非照射T-ALL白血病小鼠生存期明显延长(P<0.05)。阿霉素联合槲皮素组相对于阿霉素组SOD活性明显提高并降低MDA含量。通过转录组测序分析并经验证发现,阿霉素联合槲皮素组和槲皮素组相对于阿霉素组小鼠,Ighv1-84与Igkv6-14均低表达。阿霉素联合槲皮素组和阿霉素组相对于槲皮素组小鼠,Ms4a1、Podx1、Mecom、Sh3bgr12、Bex4、Tdrp高表达,而Crabp1低表达。结论:槲皮素对原代白血病细胞有增殖抑制作用,其抑制细胞增殖效应表现时间依赖性;槲皮素联合阿霉素后对原代白血病细胞的增殖抑制作用明显,对原代白血病细胞的增殖抑制具有协同和相加作用,且其抑制细胞增殖效应表现为浓度和时间依赖性。槲皮素联合高剂量阿霉素可显著延长非照射T-ALL白血病小鼠的生存期并降低阿霉素对于心肌的损伤。

Objective:To investigate the chemotherapeutic efficency of quercetin sensitized adriamycin. Method:CCK-8 was used to detect the inhibitory effect of different doses of adriamycin,quercetin and quercetin combined with adriamycin on the proliferation of primary leukemia cells from patients with clinically refractory acute leukemia. Quercetin,adriamycin and their combination were used to treat non-irradiated T-ALL leukemia mice to observe the changes of survival curve and myocardial injury. Result:There was no significant difference in the inhibition rate of primary leukemia cell proliferation between the adriamycin concentration group (6,0.6 and 0.06 μg/ml) and the adriamycin half-dose (3,0.3 and 0.03 μg/ml) plus quercetin (0.25 mmol/L) group at three different time points (24,48 and 72 hours). There was a significant difference in the inhibition rate of primary leukemia cell proliferation among the drug concentration groups,and the inhibition rate of primary leukemia cell proliferation was time-and concentrationdependent (r24h,a\c\e = 0.995、r48h,a\c\e = 1.000、r72h,a\c\e = 0.984、r24h,b\d\f = 0.993、r48h,b\d\f = 0.999、r72h,b\d\f = 0.960). In vivo experiments showed that the survival time of non-irradiated T-ALL leukemia mice treated with low-dose adriamycin combined with quercetin was not significantly prolonged compared with the high-dose adriamycin treatment group. The survival time of non-irradiated T-ALL leukemia mice treated with high dose of adriamycin and quercetin was significantly prolonged (P< 0.05). Compared with adriamycin group,the SOD activity in adriamycin combined with quercetin group increased significantly and the MDA content decreased. The results of transcriptome sequencing analysis showed that the expression of Ighv1-84 and Igkv6-14 in adriamycin combined quercetin group and quercetin group was lower than that in adriamycin group. The Ms4a1,Podx1,Mecom,Sh3bgr12,Bex4 and Tdrp expression in adriamycin combined quercetin group and adriamycin group were higher than that in quercetin group,while Crabp1 expression was lower. Conclusion:Quercetin can inhibit the proliferation of primary leukemia cells in a time-dependent manner. Quercetin combined with adriamycin inhibit the proliferation of primary leukemia cells significantly,and had synergistic and additive effects on the proliferation of primary leukemia cells,and the inhibiting effect of quercetin combined with adriamycin is concentrationand time-dependent. Quercetin combined with high-dose adriamycin can significantly prolong the survival time of nonirradiated T-ALL leukemia mice and reduce the myocardial damage caused by adriamycin.