盐酸小檗碱增敏柔红霉素诱导儿童急性淋巴细胞白血病原代细胞凋亡作用的研究

目的:探讨盐酸小檗碱(BBR)增敏柔红霉素(DNR)诱导儿童急性淋巴细胞白血病原代细胞(P-ALLCs)凋亡的作用及相关机制。方法:收集初诊初治的急性淋巴细胞白血病(ALL)儿童骨髓标本12例,分离单个核细胞,按活细胞数2×10^5/m L接种于含20%小牛血清1640的96孔培养板,培养4 h后按照设定DNR 4个浓度和BBR 3个浓度单药及交叉联合应用方案加入不同体积的PBS稀释DNR和BBR储备液培养干预24 h,采用cell counting kit-8 assay(CCK-8)检测DNR和BBR单药作用以及两者联合作用对P-ALLCs增殖的抑制作用。结果:DNR单药作用时抑制率为(11.56%±0.73%)^{59.60%±2.80%},随着DNR浓度的增加,P-ALLCs的增殖抑制逐渐增强,且后一浓度与前一浓度比较差异均有统计学意义(P均<0.05);BBR单药作用时抑制率为(12.39%±1.50%)^{39.75%±2.43%},随着BBR浓度的增加,P-ALLCs的增殖抑制逐渐增强,且后一浓度与前一浓度比较差异均有统计学意义(P均<0.05);DNR联合BBR时的抑制率为(18.43%±1.11%)^{89.96%±1.40%}。在4个联合用药组中,随着BBR浓度的增加,对P-ALLCs的增殖抑制逐渐增强(P均<0.05),而且每一组中DNR联合BBR后的抑制率比DNR单药作用强(P均<0.05)。结论:DNR和BBR对儿童P-ALLCs均有细胞毒性作用并成浓度依赖性。两药联合作用时BBR可以增加P-ALLCs对DNR的敏感性,该作用与浓度有一定的相关性。

Effects of Berberine Enhance on Daunorubicin-Induced Apoptosis of Primary Acute Lymphoblastic Leukemia Cells in Children

[Abstract] Objective: To probe into the effects of berberine (BBR) enhance on daunorubicin (DNR)-induced apoptosis of primary acute lymphoblastic leukemia cells (P-ALLCs) in children and the related mechanism. Methods: Bone marrow samples were collected from 12 children newly diagnosed with acute lymphoblastic leukemia (ALL), mononuclear cells were isolated, 2×105/mL living cells was inoculated in the 96-well culture plate containing 20% calf serum 1640. After culture for 4 h, different volumes of PBS were added to dilute DNR and BBR stock solutions according to the single drug and cross-combination application scheme of concentration of 4 DNR and 3 BBR for 24 h of culture intervention. Cell counting kit-8 assay (CCK-8) was used to detect the single drug effects of DNR and BBR and the combination effects on the inhibition of P-ALLCs proliferation. Results: The inhibition rate of DNR was (11.56%±0.73%) to (59.60%±2.80%), with the increase of DNR concentration, the proliferation inhibition of P-ALLCs increased, and the difference between the latter concentration and the previous concentration was statistically significant (P<0.05). The inhibition rate of BBR was (12.39%±1.50%) to (39.75%±2.43%), with the increase of BBR concentration, the proliferation inhibition of P-ALLCs increased, and the difference between the latter concentration and the previous concentration was statistically significant (P<0.05). The inhibition rate of DNR combined with BBR was (18.43%±1.11%) to (89.96%±1.40%). In the four combined groups, the proliferation inhibition of P-ALLCs increased with the increase of BBR concentration (P<0.05), and the inhibition rate of DNR combined with BBR in each group was stronger than that of DNR alone (P<0.05). Conclusion: Both DNR and BBR shows cytotoxic effects and concentration dependence on P-ALLCs in children. BBR can increase the sensitivity of P-ALLCs to DNR, and the effect is related to the concentration.