Notch信号通路在漆姑草醇提物诱导NB4细胞分化中作用  

Effect of Notch signaling pathway on differentiation of NB4 cells induced by Herba Saginae Japonicae ethanol extract

作　　者：唐文娟[1] 伏琼[1] 程勇[1] 罗世成 沈祥春[1,2] 梁冰 TANG Wen-juan;FU Qiong;CHENG Yong(Department of Pharmacology,Guizhou Medical University,Guiyang,Guizhou Province 550025,China)

机构地区：[1]贵州医科大学药理学教研室,贵州贵阳550025 [2]环境污染与疾病监控省部共建教育部重点实验室

出　　处：《中国公共卫生》2018年第8期1110-1113,共4页Chinese Journal of Public Health

基　　金：贵阳市科技计划项目(筑科合同[2017]30-9号);贵州省教育厅自然科学研究项目(黔教科2010033号);贵州省科技创新人才团队项目[黔科合平台人才(2016)5625号]

摘　　要：目的探讨Notch信号通路在漆姑草(HSJ)醇提物诱导人急性早幼粒白血病细胞(NB4)分化中的作用。方法实验设对照组和低、中、高剂量漆姑草组(30、60、120μg/m L);作用NB4细胞48 h后,采用四甲基偶氮唑蓝(MTT)比色法检测NB4细胞增殖抑制率;Wright-Giemsa染色法观察NB4细胞形态;Real time-PCR检测Jagged1、Notch1、c-myc m RNA表达;Western blot法检测Jagged1、Notch1、c-myc蛋白表达。结果漆姑草作用NB4细胞后,细胞增殖受到明显抑制;与对照组比较,漆姑草组NB4细胞形态向成熟细胞改变,可见核/浆比例减小,核形呈肾形核或分叶核;与对照组比较,30、60、120μg/m L漆姑草组NB4细胞中Jagged1 m RNA表达量[分别为(0.78±0.10)、(0.32±0.15)、(0.07±0.02)]降低,Notch1 m RNA表达量[分别为(1.27±0.14)、(1.39±0.23)、(1.53±0.30)]升高,c-myc m RNA表达量[分别为(0.88±0.05)、(0.67±0.02)、(0.47±0.16)]降低,差异均有统计学意义(均P<0.05);与对照组比较,30、60、120μg/m L漆姑草组NB4细胞中Jagged1蛋白表达量[分别为(0.95±0.10)、(0.69±0.02)、(0.59±0.01)]降低,Notch1蛋白表达量[分别为(0.56±0.08)、(0.81±0.03)、(1.33±0.14)]升高,c-myc蛋白表达量[分别为(0.82±0.14)、(0.70±0.07)、(0.65±0.20)]降低,差异均有统计学意义(均P<0.05)。结论漆姑草能诱导NB4细胞向成熟细胞方向分化,其分化诱导作用机制可能与Notch信号通路有关。Objective To study the effect of Notch signaling pathway on the differentiation of human acute promyelocytic leukemia cell line NB4 induced by Herba Saginae Japonicae ethanol extract（HSJ）. Methods The NB4 cells were treated with low-, moderate-, high-dose（30, 60, 120 μg/m L） HSJ for 48 hours and a negative control group was also established. The inhibitive rate of NB4 cell′s proliferation was detected with 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide（MTT） assay. The morphology of the NB4 cells was observed with Wright-Giemsa staining.The m RNA and protein expressions of Jagged1, Notch1, and c-myc were detected with real time-PCR and Western blot,respectively. Results Compared with that of the control group, the proliferation of HSJ-treated NB4 cells was inhibited significantly, with the observed cells′ mature morphology as decreased nucleus/cytoplasm ratio and reniform and lobulated nuclei. In comparison to those of the control group, significantly decreased m RNA expressions of Jagged1（0.78 ± 0.10, 0.32 ±0.15, and 0.07 ± 0.02） and c-myc（0.88 ± 0.05, 0.67 ± 0.02, and 0.47 ± 0.16） and significantly increased m RNA expression of Notch1（1.27 ± 0.14, 1.39 ± 0.23, and 1.53 ± 0.30） were observed for the NB4 cells treated with low-, moderate-, and highdose HSJ（P〈0.05 for all）; significantly decreased protein expressions of Jagged1（0.95 ± 0.10, 0.69 ± 0.02, and 0.59 ± 0.01）and c-myc（0.82 ± 0.14, 0.70 ± 0.07, and 0.65 ± 0.20） and significantly increased protein expression of Notch1（0.56 ± 0.08,0.81 ± 0.03, and 1.33 ± 0.14） were also observed for the NB4 cells treated with low-, moderate-, and high-dose HSJ（all P〈0.05）. Conclusion HSJ can induce mature differentiation of NB4 cells and the mechanism of the effect may be related to Notch signaling pathway.

关 键 词：漆姑草(HSJ) 人急性早幼粒白血病细胞(NB4) 诱导分化 NOTCH信号通路 

分 类 号：R114[医药卫生—卫生毒理学]