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  

Study on the Mechanism of Inducing Apoptosis of AML-M2 Cells in Vitro by Ethanol Extraction of Hedyotis Diffusa Willd.

作　　者：陈智[1] 林圣云[2] 李建新[1] 李晖[1] 熊昊[1] 聂应明 裘玫 CHEN Zhi;LIN Shengyun;LI Jianxin;LI Hui;XIONG Hao;NIE Yingming;QIU Mei(Department of Hematology,Wuhan Children’s Hospital,Tongji Medical College,Huazhong University of Science&Technology,Wuhan 430016,China;Department of Hematology,The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China;Clinical College of Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430061,China)

机构地区：[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

出　　处：《中国现代应用药学》2020年第17期2067-2072,共6页Chinese Journal of Modern Applied Pharmacy

基　　金：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)。

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。0.04,0.06,0.08mg·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。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, IkB?, p-IkB?, IKK?/?, 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.

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 

分 类 号：R965.1[医药卫生—药理学]