青黄散组分抑制BCL-2、XIAP-1、c-IAP协同诱导KG1a细胞凋亡的实验研究  

Experimental study on Qinghuang Powder through inhibition of BCL-2,XIAP-1,c-IAP synergetic induced apoptosis in KG1a cells

作　　者：吴建伟 黄建栩 郑荣 叶永斌[2] 范佳鑫 李元明 曾英坚[3] 郭坤元[4] WU Jian-wei;HUANG Jian-xu;ZHENG Rong;YE Yong-bin;FAN Jia-xin;LI Yuan-ming;ZENG Ying-jian;GUO Kun-yuan(Department of Hematology,Jiangmen Hosptial of Traditional Chinese Medicine Affiliated to Jinan University,Guangdong Province,Jiangmen 529000,China;Department of Hemoendocrinology,People′s Hosptial of Zhongshan City,Guangdong Province,Zhongshan 528400,China;.Department of Hematology,Hosptial Affiliated to Jiangxi University of Traditional Chinese Medicine,Jiangxi Province,Nanchang 330000,China;Department of Hematology,Zhujiang Hosptial,Southern Medical University,Guangdong Province,Guangzhou 510282,China)

机构地区：[1]暨南大学附属江门中医院血液病科,广东江门529000 [2]广东省中山市人民医院血液科,广东中山528400 [3]江西中医药大学附属医院血液内分泌科,江西南昌330000 [4]南方医科大学珠江医院血液科,广东广州510282

出　　处：《中国当代医药》2018年第18期10-16,共7页China Modern Medicine

基　　金：广东省江门市科技局科技计划立项项目(2016100-21)

摘　　要：目的研究中药砷复合制剂青黄散(由青黛和雄黄二药组成)组分靛玉红(Indirubin,青黛的有效成分)和二硫化二砷(Arsenic Disulfide,As_2S_2,雄黄的有效成分)对非M3型急性髓系白血病(AML)KG1a细胞株的单独和联合杀伤作用,并探讨相关杀伤作用机制。方法采用瑞氏姬姆沙染色法观察靛玉红与As_2S_2作用于KG1a细胞的形态,采用流式细胞术检测靛玉红与As_2S_2作用于KG1a细胞凋亡率,采用CCK-8检测靛玉红与As_2S_2作用于KG1a细胞抑制率,进一步用Western blot检测KG1a细胞Bcl-2、Smac、XIAP-1、c-IAP、Caspase-3蛋白的表达。结果青黄散组分靛玉红与As_2S_2单独对KG1a细胞均具有增殖抑制作用,联合用药的增殖抑制率、凋亡率作用更明显(P<0.05),Compu Syn软件分析靛玉红和As_2S_2以1∶1组合对KG1a细胞株的联合指数(CI)<0.9。联合用药在形态学上能明显促进细胞的凋亡变化,增加细胞的Caspase-3和Smac蛋白表达,抑制BCL-2、XIAP-1、c-IAP蛋白的表达。结论青黄散组分靛玉红与As_2S_2能联合抑制KG1a细胞增殖和诱导其凋亡,其机制与抑制BCL-2、XIAP-1、c-IAP蛋白表达,促进Caspase-3、Smac蛋白表达相关。本药可用于治疗常规药物抵抗的AML并为非M3型AML的治疗提供相关实验依据。Objective To study the effects of Qinghuang Powder(composed of two components of Indigo naturalis and Realgar),which the active ingredients are Indirubin and Arsenic Disulfide(As2S2),alone and in combination with cytotoxic effect on non M3 AML KG1a cells,and to investigate the mechanism of cytotoxic effect.Methods The cell morphology of KG1a cells acted on with Indirubin and As2S2 was observed by Wright Gimsa staining;the rate of cell apoptosis of KG1a cells acted on with Indirubin and As2S2 was detected by flow cytometry;CCK-8 detected for apoptosis rate of KG1a cells acted on with Indirubin and As2S2;the expression of Bcl-2,Smac,XIAP-1,c-IAP,Caspase-3 protein of KG1a cells was detected by Western blot.Results Both Indirubin and As2S2 had inhibitory effect on proliferation of KG1a cells.Compare with single dose,the combination of Indirubin and As2S2 had a higher cell proliferation inhibition rate and apoptosis rate(P<0.05).When Indirubin and As2S2 combined with 1∶1,the combined index was less than 0.9 through CompuSyn,which could also influenced the apoptosis highly,increased the expression of Caspase-3 and Smac protein,and inhibit the expression of BCL-2,XIAP-1 and c-IAP protein.Conclusion The Indirubin and As2S2,the ingredient of Qinghuang Powder,can inhibit the proliferation of KG1a cells and induce its apoptosis,the mechanism may be related to inhibition of the expression of BCL-2,XIAP-1,c-IAP protein,promote the expression of Caspase-3,Smac protein.Qinghuang powder can be used for the treatment of AML of conventional drug resistance and provides the relevant experimental basis for the treatment of non-M3 type AML.

关 键 词：青黄散 KG1a 二硫化二砷 靛玉红 凋亡 

分 类 号：R33[医药卫生—人体生理学]