艾迪注射液联合地塞米松对急性淋巴白血病细胞Jurkat的增殖抑制及机制研究  

Proliferation inhibition of acute lymphoblastic leukemia Jurkat cells by aidi injection in combination with dexamethasone and its mechanism

作　　者：杨坤 刘琴[1,2,3] 韦学耐 宋晶睿[1,3] 饶青 黄裕兵 李艳梅 YANG Kun;LIU Qin;WEI Xuenai;SONG Jingrui;RAO Qing;HUANG Yubing;LI Yanmei(Pharmacology and Biological Activity Research Center,Guizhou Natural Products Research Center,Guiyang 550014,Guizhou,China;School of Pharmacy,Guizhou Medical University,Guiyang 550004,Guizhou,China;Provincial and Ministerial State Key Laboratory for Functions and Applications of Medicinal Plants,Guizhou Medical University,Guiyang 550014,Guizhou,China)

机构地区：[1]贵州省天然产物研究中心药理与生物活性研究中心,贵州贵阳550014 [2]贵州医科大学药学院,贵州贵阳550004 [3]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,贵州贵阳550014

出　　处：《贵州医科大学学报》2025年第1期9-17,共9页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(81960546);贵州省科技计划项目(黔科合平台人才[2020]5008)。

摘　　要：目的探讨艾迪注射液(aidi injection,ADI)联合地塞米松(dexamethasone,DEX)对急性淋巴白血病细胞Jurkat增殖抑制的影响及机制。方法MTT法检测不同浓度的DEX对白血病细胞Jurkat、CEM-C1细胞增殖的影响,并通过流式细胞术检测DEX在48 h内对Jurkat细胞凋亡的影响;ADI单用或联合DEX处理细胞24、48 h,采用MTT法检测细胞存活率;流式细胞术检测ADI单用或联合DEX对Jurkat细胞周期和细胞凋亡的影响;Western blot法检测ADI单用或联合DEX处理Jurkat细胞后各组中半胱氨酸蛋酶家族蛋白(Caspase-9、Cleaved caspase-9、Caspase-3及Cleaved caspase-3)、DNA修复酶(PARP、Cleaved-PARP)、Bcl-2家族凋亡蛋白(Bcl-2、Bcl-XL、BIM和BAD)、细胞周期相关蛋白(C-Myc、Cyclin E1和CDK2)、糖皮质激素受体(glucocorticoid receptor,GR)、磷酸化-GR(P-GR)、磷酸化-Janus激酶2(P-JAK2)、磷酸化-信号转导与激活因子3(P-STAT3)及磷酸化蛋白激酶B(P-AKT)的蛋白表达。结果相对于CEM-C1细胞,Jurkat细胞对DEX不敏感,仅在高浓度的DEX(200μmol/L)能够诱导Jurkat较少的细胞凋亡(P<0.05);ADI能够以浓度依赖的方式抑制急性白血病Jurkat细胞的增殖(P<0.05);与对照组相比,ADI联合DEX能更明显的抑制Jurkat细胞增殖(P<0.05),细胞凋亡率显著增加(P<0.05),使细胞周期阻滞在G1期(P<0.05);与对照组相比,Western blot检测显示,ADI联合DEX能显著上调Cleaved caspase-9、Cleaved caspase-3、Cleaved-PARP、BIM、BAD和P-GR蛋白的表达(P<0.05),显著下调C-Myc、Cyclin E1、CDK2、GR、P-JAK2、P-STAT3和P-AKT蛋白表达(P<0.05)。结论ADI联合DEX能够增强抑制Jurkat细胞增殖的活性,使细胞周期阻滞于G1期,通过诱导细胞凋亡杀伤Jurkat细胞,其机制可能是通过激活GR的表达,影响JAK2/STAT3/AKT通路克服Jurkat细胞的地塞米松耐药。Objective To investigate proliferation inhibition of acute lymphoblastic leukemia(ALL)Jurkat cells by aidi injection(ADI)in combination with dexamethasone(DEX)and its mechanism.Methods MTT assay was applied to examine the effect of DEX at different concentrations on Jurkat and CEM-C1 leukemia cell proliferation.Flow cytometry was used to examine the effect of DEX within 48 h on Jurkat cell apoptosis.Cells were treated with ADI alone or in combination with DEX for 24 and 48 h,respectively,and cell survival rate was detected by MTT assay.Flow cytometry was used to examine the effects of ADI alone or DEX on Jurkat cell cycle and apoptosis.Western blot was applied to detect protein expressions of caspase family members(Caspase-9,Cleaved caspase-9,Caspase-3,and Cleaved caspase-3),DNA repair enzymes[poly ADP-ribose polymerase(PARP)and cleaved-PARP],B-cell lymphoma-2(Bcl-2)family members[Bcl-2,B-cell leukemia/lymphoma xL(Bcl-XL),Bcl-2 interacting mediator of cell death(BIM),and Bcl-2 asociated death promoter(BAD)],cell cycle-related proteins[myelocytomatosis viral oncogene homolog(C-Myc),cyclin-dependent kinase E1(Cyclin E1),and cyclin-dependent kinase 2(CDK2)],glucocorticoid receptor(GR),phospho-GR(P-GR),phospho-Janus kinase 2(P-JAK2),phosphorylation signal transducer and activator of transcription 3(P-STAT3),and phosphorylation protein kinase B(P-AKT)after Jurkat cells were treated with ADI alone or or in combination with DEX.Results Jurkat cells were insensitive to DEX relative to CEM_C1 cells.Only high concentration of DEX(200μmol/L)could induce apoptosis of Jurkat cells(P<0.05).ADI inhibited Jurkat cell proliferation in a concentration-dependent manner(P<0.05).When compared to control group,ADI combined with DEX obviously inhibited Jurkat cell proliferation(P<0.05),significantly increased apoptosis(P<0.05)and induced cell cycle arrest in G1 phase(P<0.05).When compared to control group,Western blot results showed that ADI combined with DEX significantly upregulated the protein expressions of Cleaved caspase-9,Cleav

关 键 词：艾迪注射液 地塞米松 JURKAT细胞 耐药 

分 类 号：R733.7[医药卫生—肿瘤]