漆姑草醇提物对人急性早幼粒白血病细胞的诱导分化作用

目的： 探讨漆姑草醇提物（HSJ）对人急性早幼粒白血病细胞（NB4）的诱导分化作用。方法： 以NB4细胞为研究对象，设HSJ低（30 μg/mL）、中（60 μg/mL）、高（120 μg/mL）剂量组和阴性对照组，共4组；HSJ作用NB4细胞48 h后，采用透射电镜法观察NB4细胞形态；硝基四唑氮（NBT）还原实验检测NB4细胞分化能力；流式细胞术检测NB4细胞周期分布；流式细胞术检测NB4细胞表面分化抗原CD14和CD11b细胞的表达率；实时荧光定量PCR （qPCR）检测c-fos mRNA表达；Western blot检测c-fos蛋白表达。结果： 与对照组比较，3个不同剂量HSJ处理组NB4细胞胞核均缩小，胞浆呈空泡状，核形呈肾形或蚕豆形；各HSJ处理组NB4细胞的NBT还原率均高于对照组，差异有统计学意义（P < 0.05）；HSJ处理组NB4细胞中G2期细胞比例均高于对照组，而S期细胞比例均低于对照组，差异有统计学意义（P < 0.05）；HSJ处理组CD14和CD11b阳性细胞表达率均高于对照组（P < 0.05）；HSJ低剂量组cfos mRNA和蛋白表达与对照组比较，差异均无统计学意义（P > 0.05），HSJ中、高剂量组c-fos mRNA和蛋白表达均高于对照组，差异均有统计学意义（P < 0.05）。结论： HSJ可能诱导NB4细胞向成熟粒细胞方向分化。

Induction of differentiation of human acute promyelocytic leukemia cells by an ethanol extract from Herba Saginae Japonicae

OBJECTIVE: To study cellular changes during induction of differentiation of human acute promyelocytic leukemia cells (NB4) by an alcohol extract from Herba Saginae Japonicae (HSJ). METHODS: NB4 cells in culture were were organized into four groups:HSJ low dose (30 μg/mL), HSJ middle dose (60 μg/mL), HSJ high dose (120 μg/mL) and negative control groups. Cells in the experimental groups were treated with HSJ for 48 h and their cellular morphology was observed by transmission electron microscopy. nitrotriazole nitrogen (NBT) reduction test was used to detect the differentiation ability of NB4 cells. The distribution of NB4 cell cycles and expression rates of CD14 and CD11b cells were detected using flow cytometry. Expression of c-fos mRNA was detected using quantitative real-time PCR(qPCR). Expression of cfos protein was detected using the Western blot method. RESULTS: Compared with the control group, the nuclei of HSJ-treated cells, in the three different doses, were reduced and changed into kidney-shape, and the cytoplasm was vacuolated. The NBT test showed that the reduction rate of the treated cells were significantly higher than that in the control group (P < 0.05). The proportions of G2 phase cells in the treated groups were significantly higher than that in the control group, while the proportion of S phase cells were significantly lower (P < 0.05). The expression rates of CD14 and CD11b positive cells in the treated groups were significantly higher than that in the control group (P < 0.05). In addition, expression rates of c-fos mRNA and c-fos proteins in the middleand the high-dose groups were significantly higher than that in the control group (P < 0.05) but no difference for the low-dose group. CONCLUSION: HSJ may induce NB4 cells to differentiate into mature granulocytes.