当归多糖调节骨髓造血微环境治疗再生障碍性贫血的机制  

Mechanism by which Angelica sinensis polysaccharide regulates bone marrow hematopoietic microenvironment for aplastic anemia

作　　者：付佳琪 陈修保[2] 崔兴[3] 陈泽涛[2] Fu Jiaqi;Chen Xiubao;Cui Xing;Chen Zetao(School of Traditional Chinese Medicine,Shandong University of Traditional Chinese Medicine,Jinan 250013,Shandong Province,China;Department of Gerontology,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250013,Shandong Province,China;Center of Oncology,Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250001,Shandong Province,China)

机构地区：[1]山东中医药大学中医学院,山东省济南市250013 [2]山东中医药大学附属医院老年病科,山东省济南市250013 [3]山东中医药大学第二附属医院肿瘤中心,山东省济南市250001

出　　处：《中国组织工程研究》2025年第1期44-51,共8页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金面上项目(ZR2020MH354),项目负责人:陈泽涛。

摘　　要：背景:如何改善造血微环境是目前再生障碍性贫血治疗的热点问题。目的:结合GEO测序分析、网络药理学与实验验证当归多糖改善骨髓造血微环境治疗再生障碍性贫血的作用机制。方法:借助GEO数据库获得了再生障碍性贫血相关差异表达基因并进行GO和GSEA分析。结合文献与PubChem、SwissTargetPrediction、PharmMapper数据库获取当归多糖活性成分和靶点。取交集靶点后利用STRING和Cytoscape构建蛋白相互作用网络,并进行GO和KEGG富集分析。构建再生障碍性贫血小鼠模型,利用血细胞分析仪、流式细胞术、ELISA、免疫组化、免疫荧光染色、Western blot方法验证当归多糖对再生障碍性贫血的疗效与作用机制。结果与结论:(1)共筛选出834个差异表达基因,参与细胞发育、造血、髓系细胞分化等生物学过程;(2)检索得到当归多糖相关347个靶点并筛选出77个潜在治疗基因,其中VEGFA、EGLN1、BCL2、干扰素γ、白细胞介素2、白细胞介素4、白细胞介素6等血管生成、凋亡及免疫相关因子度值显著;(3)KEGG通路富集分析治疗靶点主要富集在Th17细胞分化、NK相关细胞毒性、细胞黏附因子等干扰素γ、白细胞介素2、白细胞介素4相关信号通路;(4)动物实验表明,当归多糖能够显著改善再生障碍性贫血小鼠骨髓造血,增加外周血细胞及骨髓单个核细胞计数,提高小鼠存活率;与模型组相比,当归多糖组小鼠Th1/Th2细胞比例显著下降(P<0.01),干扰素γ水平显著降低(P<0.01),白细胞介素4水平升高(P<0.05),VEGFA表达显著升高(P<0.01),EGLN1表达显著降低(P<0.01),骨髓单个核细胞凋亡率、活性氧水平显著降低(P<0.01),Cleaved Caspase-3蛋白表达显著增高(P<0.01),Bcl-2/Bax比值显著降低(P<0.01);(5)结果表明:当归多糖能够通过调节异常T细胞亚群,促进血管生成以改善造血微环境,并抑制骨髓单个核细胞凋亡,以改善再生障碍性贫血小鼠的造血功BACKGROUND:How to improve the hematopoietic microenvironment is a hot topic in the treatment of aplastic anemia.OBJECTIVE:To explore the action mechanism of Angelica sinensis polysaccharide in the treatment of aplastic anemia by combining GEO sequencing analysis,network pharmacology,and experimental validation.METHODS:Aplastic anemia-related differentially expressed genes were obtained from GEO database,and gene ontology and gene set enrichment analysis were performed.The active components and targets of Angelica sinensis polysaccharide were obtained by combining the literature with PubChem,SwissTargetPrediction,and PharmMapper databases.After the intersection targets were taken,STRING and Cytoscape were used to construct proteinprotein interaction network,and gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis was performed.Mouse model of aplastic anemia was established,and the effect and action mechanism of Angelica sinensis polysaccharide on aplastic anemia were verified by blood cell analyzer,flow cytometry,ELISA,immunohistochemistry,immunofluorescence,and western blot assay.RESULTS AND CONCLUSION:(1)A total of 834 differentially expressed genes were screened,which were involved in biological processes such as cell development,hematopoiesis,and myeloid cell differentiation.(2)347 targets related to Angelica sinensis polysaccharide were retrieved and 77 potential therapeutic genes were screened.Among them,the degree values of angiogenic,apoptotic,and immune-related factors such as VEGFA,EGLN1,Bcl-2,interferon-γ,interleukin-2,interleukin-4,and interleukin-6 were significant.(3)KEGG pathway enrichment analysis revealed that the therapeutic targets were mainly enriched in Th17 cell differentiation,NK-related cytotoxicity,cell adhesion factors such as interferon-γ,interleukin-2,and interleukin-4 related signaling pathways.(4)Animal experiments showed that Angelica sinensis polysaccharide significantly improved bone marrow haematopoiesis,increased peripheral blood cell,and bone marrow si

关 键 词：当归多糖 再生障碍性贫血 TH1/TH2 骨髓造血微环境 细胞凋亡 

分 类 号：R453[医药卫生—治疗学] R318[医药卫生—临床医学] R551