基于网络药理学与实验验证探究生血宁治疗β-地中海贫血作用机制
Exploring the mechanism of action of Shengxuening for the treatment of β-thalassemia based on network pharmacology and experimental validation
吴茜 1李天航 2吴金虎 2张恩景2
作者信息
- 1. 武汉大学同仁医院健康体检中心,湖北武汉 430060
- 2. 武汉大学同仁医院药学部,湖北武汉 430060
- 折叠
摘要
目的:探究生血宁治疗β-地中海贫血的机制.方法:根据文献数据,利用相应数据库检索生血宁主要成分蚕砂活性成分及相关靶点、β-地中海贫血相关的疾病靶点.对药物治疗靶点及疾病靶点进行交集分析,得核心交集靶点.利用核心交集靶点信息进行蛋白互作网络分析、KEGG及GO富集分析.使用不同浓度(100,200,400,800μg·mL-1)生血宁培养K562细胞.CCK-8法检测细胞活力,联合使用Elisa、Western blot检测K562细胞胎儿血红蛋白表达水平,使用实时荧光定量分析法(Real-Time PCR)检测Gγ、Aγ mRNA表达情况.使用P38、ERK、JNK特异性抑制剂与生血宁联合处理K562细胞.Elisa检测细胞HbF 含量,Western blot 检测细胞 HbF、P-P38/P38、P-ERK/ERK、P-JNK/JNK 表达水平,q-PCR 法检测 GγAγ mRNA 表达情况.结果:网络药理学确定生血宁可以通过MAPK信号通路治疗β-地中海贫血.分子实验确定生血宁有效给药剂量为400μg·mL-1,有效给药时间为48h.生血宁通过激活P38MAPK信号通路,促进Gγ、Aγ mRNA表达,细胞内HbF含量增加.与对照组相比,实验组HbF含量及Gγ、Aγ水平均显著升高.结论:生血宁通过P38MAPK信号通路,激活γ-珠蛋白基因,促进胎儿血红蛋白合成.
Abstract
OBJECTIVE To explore the treatment of Shengxuening β-the mechanism of thalassemia.METHODS Accord-ing to the literature data,the active components and related targets of the main components of shengxuening,silkworm sand,andβ-thalassemia related disease targets were searched by using the corresponding database.Cross-tabulation analysis of drug thera-peutic targets and disease targets was performed to obtain core cross-tabulation targets.The core intersection target information was used for protein interaction network analysis,KEGG and GO enrichment analysis.K562 cells were cultured with different concentrations(100,200,400,800 μg·mL-1)of raw hematoxylin.cell viability was measured by CCK-8 method,and fetal hemo-globin expression level of K562 cells was measured by combined Elisa and Western blot,and real-time fluorescence quantitative analysis(Real-Time PCR)to detect Gγ and Aγ mRNA expression.K562 cells were treated with P38,ERK,JNK specific inhibi-tors in combination with biotin.Elisa was used to detect cellular HbF content,Western blot to detect cellular HbF,P-P38/P38,P-ERK/ERK,P-JNK/JNK expression levels,and q-PCR to detect Gγ,Ay mRNA expression.RESULTS Network pharmacol-ogy determined that Shengxuening can treat β-thalassemia through the MAPK signaling pathway.Molecular experiments deter mined that Shengxuening was effectively administered at a dose of 400 μg·mL-1 for 48 h.Shengxuening promoted Gγ and AγmRNA expression and increased intracellular HbF content through activation of the P38 MAPK signaling pathway,.Compared with the control group,HbF content and Gγ and Ay levels were significantly increased in the experimental group.CONCLUSION Shengxuening promotes fetal hemoglobin synthesis by activating the gamma-cadherin gene through the P38 MAPK signaling pathway.
关键词
地中海贫血/生血宁/胎儿血红蛋白/MAPK信号通路/网络药理学Key words
thalassemia/Shengxuening/fetal hemoglobin/MAPK signaling pathway/network pharmacology引用本文复制引用
出版年
2024
NETL
NSTL
维普
万方数据