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引用本文:陈智,林圣云,李建新,李晖,熊昊,聂应明,裘玫.白花蛇舌草乙醇提取物体外诱导人AML-M2白血病细胞凋亡机制研究[J].中国现代应用药学,2020,37(17):2067-2072.
CHEN Zhi,LIN Shengyun,LI Jianxin,LI Hui,XIONG Hao,NIE Yingming,QIU Mei.Study on the Mechanism of Inducing Apoptosis of AML-M2 Cells in Vitro by Ethanol Extraction of Hedyotis Diffusa Willd.[J].Chin J Mod Appl Pharm(中国现代应用药学),2020,37(17):2067-2072.
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白花蛇舌草乙醇提取物体外诱导人AML-M2白血病细胞凋亡机制研究
陈智,林圣云,李建新,李晖,熊昊,聂应明,裘玫
1.华中科技大学同济医学院附属武汉儿童医院血液肿瘤科, 武汉 430016;2.浙江中医药大学附属第一医院血液科, 杭州 310006;3.湖北中医药大学中医临床学院, 武汉 430061
摘要:
目的 观察白花蛇舌草乙醇提取物(ethanol extract of Hedyotis diffusa Willd.,EEHDW)对AML-M2白血病Kasumi-1细胞增殖和凋亡的影响机制。方法 采用MTT比色法、Hoechest荧光染色检测测定EEHDW作用后Kasumi-1细胞增殖和凋亡的情况,Western blotting法检测Kasumi-1细胞中Bcl-2、Bax、caspase-3、caspase-9、Cyto-C、P65、p-P65、IkBα、p-IkBα、IKKα/β、C-myc以及AML1-ETO的蛋白水平。结果 EEHDW抑制急性髓系白血病Kasumi-1细胞增殖,并具有时间和浓度依赖性。0.04,0.06,0.08 mg·mL-1的EEHDW能诱导细胞凋亡(P<0.05或P<0.01)。EEHDW可以显著上调Cyto-C、cleaved PARP、cleaved caspase-3、cleaved caspase-9以及Bax的表达水平,而降低Bcl-2、C-myc和AML1-ETO蛋白的表达,同时EEHDW能够浓度依赖性地增加p-P65、p-IkBα的蛋白表达。结论 EEHDW诱导Kasumi-1细胞凋亡一方面是通过调节Bax/Bcl-2的表达影响线粒体途径,另外一方面还与激活NF-кB信号通路有关,在这个过程中同时抑制原癌基因C-myc和AML1-ETO融合基因的表达。
关键词:  白花蛇舌草乙醇提取物  Kasumi-1细胞  凋亡  NF-kB信号通路
DOI:10.13748/j.cnki.issn1007-7693.2020.17.004
分类号:R965.1
基金项目:湖北省卫生计生委面上项目(WJ2017M195);武汉市中青年医学骨干人才培养计划(武卫生计生[2014]77号);武汉市卫计委临床医学科研项目(WX13B19)
Study on the Mechanism of Inducing Apoptosis of AML-M2 Cells in Vitro by Ethanol Extraction of Hedyotis Diffusa Willd.
CHEN Zhi1,2, LIN Shengyun3, LI Jianxin1,2, LI Hui1,2, XIONG Hao1,2, NIE Yingming1,2, QIU Mei4
1.Department of Hematology, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science &2.Technology, Wuhan 430016, China;3.Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China;4.Clinical College of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan 430061, China
Abstract:
OBJECTIVE To observe the effect of ethanol extract of Hedyotis diffusa Willd. (EEHDW) on the proliferation and apoptosis of AML-M2 cells Kasumi-1 and its mechanism. METHODS MTT colorimetry and Hoechest staining were used to detect the effect of EEHDW on the proliferation and apoptosis of Kasumi-1 cells. Western blotting was used to detect the protein levels of Bcl-2, Bax, caspase-3, caspase-9, Cyto-C, p65, p-P65, IkBa, p-IkBa, IKKa/b, C-myc and AML1-ETO in Kasumi-1 cells. RESULTS EEHDW inhibited the proliferation of Kasumi-1 cells. The 0.04, 0.06 and 0.08 mg·mL-1 of EEHDW could induce apoptosis(P<0.05 or P<0.01). It could significantly up regulate the expression of Cyto-C, cleaved PARP, cleaved caspase-3, cleaved caspase-9 and Bax, but decreased the expression of Bcl-2, C-myc and AML1-ETO. Meanwhile, EEHDW could increase the expression of p-P65 and p-IKBα in a concentration dependent manner. CONCLUSION EEHDW induce apoptosis of Kasumi-1 cells not only related to the regulation of Bax/Bcl-2 expression, but also relate to the activation of NF-кB signal pathway, which inhibits the expression of C-myc and AML1-ETO fusion gene.
Key words:  ethanol extract of Hedyotis diffusa Willd.  Kasumi-1 cell  apoptosis  NF-kB signal pathway
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