摘要
目的 研究青黄散治疗骨髓增生异常综合征(Myelodysplasticsyndrome, MDS)的潜在分子靶点及作用机制。方法 研究纳入15例MDS患者,在治疗前以及青黄散治疗6个月后提取骨髓样本,以转录组测序检测治疗前后骨髓单个核细胞基因的差异表达,并通过基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析对差异表达基因进行功能富集分析。利用Cytoscape 3.9.1软件构建差异表达基因蛋白质-蛋白质相互作用网络(PPI);为了验证体内结果,选取MDS细胞株MUTZ-1作为实验对象作体外实验,以青黄散主要有效成分硫化砷为干预手段,使用实时定量PCR验证测序结果。结果 转录组分析证实,与治疗前比较,经青黄散治疗6个月后MDS患者骨髓有核细胞有2841个差异表达基因,包括1892上调基因和949个下调基因,GO功能富集分析发现这些差异表达基因参与了组蛋白修饰这一与MDS高度相关的生物过程。KEGG分析发现,HIF-1、TNF-α及VEGF等信号通路可能是青黄散治疗MDS的潜在通路。根据GO富集分析结果,组蛋白修饰差异表达基因被用于构建PPI蛋白互作网络并进行核心基因的筛选,进一步体外验证实验发现,硫化砷干预MDS细胞株后EP300、HDAC7与SETD2较对照组显著上调,HDAC6显著下调,与测序结果一致。结论 组蛋白修饰是青黄散治疗MDS的重要作用机理,可能涉及EP300、HDAC6、HDAC7与SETD2等分子靶点。
Objective To explore the potential molecular target and mechanism of Qinghuang Powder in the treat-ment of myelodysplastic syndrome(MDS).Methods Fifteen MDS patients were enrolled in this study.Bone marrow sam-ples were extracted before treatment and 6 months after treatment with Qinghuang Powder,and RNA-seq was performed to detect the differentially expressed genes(DEGs)in bone marrow mononuclear cells before and after treatment.Gene ontolo-gy(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed for the DEGs.Cytoscape 3.9.1 was employed to build the protein-protein interaction(PPI)network of DEGs.Furthermore,the MDS cell line MUTZ-1 was treated with arsenic sulfide,the main effective component of Qinghuang Powder,and then real-time quantitative PCR was employed to verify the RNA-seq results.Results A total of 2841 DEGs were identified by RNA-seq in bone marrow nucleated cells of MDS patients treated with Qinghuang Powder for 6 months,including 1892 up-regulated genes and 949 down-regulated genes.GO annotation showed that these DEGs were mainly involved in his-tone modification highly associated with MDS.KEGG pathway enrichment indicated that Qinghuang Powder treated MDS via the hypoxia-inducible factor-1(HIF-1),tumor necrosis factor-alpha(TNF-α),and vascular endothelial growth factor(VEGF)signaling pathways.According to GO annotation results,the DEGs associated with histone modification were used to construct the PPI network,from which the core genes were screened out.The MDS cells treated with arsenic sulfide showed significantly up-regulated expression of E1A binding protein p300(EP300),histone deacetylase 7(HDAC7),and SET domain-containing protein 2(SETD2)and down-regulated expression of HDAC6,which was consistent with the se-quencing results.Conclusion Histone modification is an important step of Qinghuang Powder in the epigenetic regulation of MDS patients.EP300,HDAC6,HDAC7,and SETD2 may be the key targets of Qinghuang Powder in the treatment of MDS.
作者
凌志明
付中学
王德秀
杨秀鹏
杜宇
谷晓丽
麻柔
许勇钢
LING Zhi-ming;FU Zhong-xue;WANG De-xiu;YANG Xiu-peng;DU Yu;GU Xiao-li;MA Rou;XU Yong-gang(Department of Hematology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091)
出处
《世界中西医结合杂志》
2023年第4期698-704,共7页
World Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(81774140)
中国中医科学院科技创新工程(CI2021A01706)。
