龙群,肖潇,宋晶睿,饶青,苑春茂,何志旭,李艳梅
LONG Qun,XIAO Xiao,SONG Jingrui,RAO Qing,YUAN Chunmao,HE Zhixu,LI Yanmei

• 1:贵州医科大学基础医学院免疫教研室&组织工程与干细胞实验中心
• 2:贵州医科大学省部共建药用植物功效与利用国家重点实验室
• 3:贵州省中国科学院天然产物化学重点实验室

摘要(Abstract)：

目的:探讨Flavaglines类化合物对人白血病HEL细胞的作用及机制。方法:取HEL细胞培养至对数生长期,以DMSO组为空白对照组,采用过四唑盐(MTT)法检测阳性对照药阿霉素(0. 100、0. 050、0. 030、0. 010及0. 006μmol/L)、化合物M8 (0. 20、0. 10、0. 05、0. 03及0. 01μmol/L)及M9 (2. 50、1. 25、0. 63、0. 30及0. 16μmol/L)对HEL细胞作用72 h后的吸光度值(OD)并计算抑制率和半抑制浓度(IC50)值;采用流式细胞仪检测M8(0. 20、0. 05及0. 02μmol/L)和M9(1. 00、0. 50及0. 25μmol/L)对HEL细胞作用24、48及72 h后的细胞凋亡率;采用Western blot分析M8(0. 20μmol/L)和M9(1. 00μmol/L)对HEL细胞作用24 h后B淋巴细胞瘤-2(Bcl-2)、Caspase3、Cleave-Caspase3、磷酸化信号传导转录激活蛋白3(p-Stat3)及c-Myc蛋白表达的变化。结果:MTT结果显示,阿霉素及化合物M8、M9抑制HEL细胞增殖的抑制率均随浓度的增加而增高(P <0. 05或P<0. 01),IC50值分别为(0. 06±0. 001)μmol/L及(0. 02±0. 010)μmol/L、(0. 21±0. 060)μmol/L;流式细胞仪结果显示,化合物M8和M9促进HEL细胞的凋亡率随浓度的增加而增高(P <0. 05或P <0. 01); Western blot结果显示,与DMSO组比较,M8和M9组HEL细胞的Bcl-2、Caspase3、p-Stat3及c-Myc蛋白表达减少(P <0. 05或P <0. 01),Cleave-Caspase3蛋白表达增加(P <0. 01)。结论:Flavaglines类化合物M8和M9可通过影响Stat3信号通路而促进HEL细胞发生凋亡。
Objective: To investigate the effect and mechanism of flavaglines on human leukemia HEL cells. Methods: Compounds M8 and M9 are flavaglines. Based on the treatment,HEL cells at logarithmic growth phase were treated with DMSO as blank group,treated with doxorubicin( 0. 100,0. 050,0. 030,0. 010 and 0. 006 μmol/L) as positive group,treated with compound M8( 0. 20,0. 10,0. 05,0. 03 and 0. 01 μmol/L) as M8 group and compound M9( 2. 50,1. 25,0. 63,0. 30 and0. 16 μmol/L) as M9 group. At 72 h after the above treatment,MTT assay was used to determine OD values. Cell growth inhibition rates and IC50 were calculated. Flow cytometry was used to detect cellular apoptosis induced by M8 and M9. Western blot was used to detect the protein levels of B lymphoma-2( Bcl-2),Caspase3,Cleaved-Caspase3,phospho-Stat3 and c-Myc. Results: The MTT assay showed that the cell growth inhibitory rates of doxorubicin,compounds M8 and M9 were increased in a dose-dependent manner( P < 0. 05 or P < 0. 01),and their IC50 values were( 0. 06 ±0. 001) μmol/L And( 0. 02 ± 0. 010) μmol/L,( 0. 21 ± 0. 060) μmol/L,respectively. Flow cytometry analysis showed that compound M8 and M9 induced cellular apoptosis in a dose-dependent manner( P < 0. 05 or P < 0. 01). Western blot analysis revealed that compared with the DMSO group,both compounds M8 and M9 downregulated the expression levels of Bcl-2,Caspase3,phospho-Stat3 and c-Myc( P < 0. 05 or P < 0. 01),while upregulated the expression levels of cleaved Caspase3( P <0. 01). Conclusion: Flavaglines M8 and M9 can promote the apoptosis of HEL cells by affecting the Stat3 signaling pathway.

关键词(KeyWords)： 白血病;细胞凋亡;环戊烷苯啶呋喃类化合物;HEL细胞;抑制增殖;信号传导转录激活蛋白3
leukemia;apoptosis;cyclopentane benzidine furans;HEL cell;proliferation inhibition;signal transducer and activator of transcription 3(Stat3)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81872772、81960546、U1812403-4);; 贵州省科技计划项目[黔科合平人才(2019)5406、QKH20181409、QKH20192762、QKH20205008]

作者(Author): 龙群,肖潇,宋晶睿,饶青,苑春茂,何志旭,李艳梅
LONG Qun,XIAO Xiao,SONG Jingrui,RAO Qing,YUAN Chunmao,HE Zhixu,LI Yanmei

DOI: 10.19367/j.cnki.2096-8388.2020.05.002

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