摘要
目的研究紫草素(shikonin,SK)通过调控内质网应激通路诱导急性髓系白血病早幼粒细胞分化的作用机制。方法使用NOD/SCID小鼠,通过尾静脉注射HL-60细胞构建白血病模型,评价紫草素对于白血病动物的治疗效果,细胞水平利用CCK-8法检测HL-60细胞活性,Giemsa染色检测细胞核形态、硝基蓝四氮唑(NBT)还原试验、ER-Tracker Red染色,探究紫草素调控内质网应激的作用。通过转录组测序技术筛选差异表达基因,结合RT-qPCR和Western blotting进行进一步机制探究。结果紫草素干预后,小鼠各脏器炎性细胞浸润的病理状态得到改善。转录组测序揭示了89个与内质网应激相关的差异表达基因,主要涉及蛋白质加工等信号通路,其中DDIT3和CEBPA为关键靶点。SK以剂量依赖性方式抑制HL-60细胞增殖,促进核形态变化,增强NBT还原活性和内质网荧光,同时改变相关基因和蛋白的表达。SK显著下调pdi和perk基因及其蛋白表达,上调cebpa、grp 78和ire1基因及其蛋白表达。使用内质网应激抑制剂牛磺熊去氧胆酸(tauroursodeoxycholic acid,TUDCA)处理后,HL-60细胞的核分裂和NBT还原能力受到抑制,相关基因和蛋白表达也相应减少。结论SK能够诱导细胞内质网应激,上调IRE1表达,抑制PDI蛋白表达,并促进CEBPA蛋白表达上调,从而启动HL-60细胞的分化过程。
Objective To investigate the molecular mechanism by which Shikonin(SK)regulates the endoplasmic reticulum stress pathway to induce the differentiation of promyelocytes in acute myeloid leukemia.Methods A leukemia model of NOD/SCID mice was established.Differentially expressed genes were screened by transcriptome sequencing.The activity of HL-60 cells was measured by the CCK-8 method.Detection and analysis were performed using techniques such as Giemsa staining,nitroblue tetrazolium(NBT)reduction test,ER-Tracker Red detection,PCR,and Western blotting.Results After SK intervention,the pathological state of inflammatory cell infiltration in various organs of the mice was improved.Transcriptome sequencing identified 89 endoplasmic reticulum-related differentially expressed genes,which were involved in pathways such as protein processing.DDIT3 and CEDPA were key targets.At the cellular level,SK inhibited the proliferation of HL-60 cells in a dose-dependent manner,promoted changes in nuclear morphology,enhanced NBT reduction activity and endoplasmic reticulum fluorescence,and altered the expression of related genes and proteins.SK significantly down-regulated pdi and perk,and up-regulated cebpa,grp78,and ire1 genes and their corresponding proteins.After treatment with the endoplasmic reticulum stress inhibitor TUDCA,the nuclear division and NBT reduction ability of HL-60 cells were inhibited,and the expression of related genes and proteins was also inhibited.Conclusion SK can induce cellular endoplasmic reticulum stress(ERS),up-regulate the expression of IRE1,inhibit the expression of PDI protein,and promote the up-regulation of CEBPA protein expression,thus initiating the differentiation process of HL-60 cells.
作者
昌潇
陈鹏
马雨贤
张波
CHANG Xiao;CHEN Peng;MA Yuxian;ZHANG Bo(School of Pharmacy,Shihezi University,Shihezi,Xinjiang 832000,China;Key Laboratory of Xinjiang Endemic Phytomedicine Resources,Ministry of Education School of Pharmacy Ministry of Education,Shihezi University,Shihezi,Xinjiang 832000,China;School of Pharmacy/Sichuan Antimicrobial Industry Research Institute,Chengdu University,Chengdu,Sichuan 610000,China)
出处
《石河子大学学报(自然科学版)》
北大核心
2025年第5期588-595,共8页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(U1603122),四川省天府峨眉创新领军人才项目(1811)。
关键词
紫草素
内质网应激
急性髓系白血病
细胞分化
转录组测序
shikonin
endoplasmic reticulum stress
acute myeloid leukemia
cell differentiation
transcriptome sequencing
