基于网络药理学探讨丹蒌片治疗糖尿病心肌病的作用机制及RAGE/NF-κB/NLRP3信号通路验证

Study on mechanism of action of Danlou tablets in treating diabetic cardiomyopathy based on network pharmacology and validation of RAGE/NF-κB/NLRP3 signaling pathway

目的应用综合药理学策略系统评价丹蒌片的药理作用及分子机制,为其用于糖尿病心肌病(diabetic cardiomyopathy,DCM)的治疗提供理论基础。方法从中药系统药理学分析平台中挖掘丹蒌片化学成分及作用靶点,建立丹蒌片“成分-靶点”相互作用网络;通过数据库筛选DCM所有作用靶点,构建“成分-疾病靶点”作用关系图,筛选其核心靶点,对核心靶点进行基因本体论分析及相关通路富集。以丹蒌片处理DCM大鼠模型,验证预测的重要信号通路。结果从丹蒌片的10种重要配方中,共筛选出化合物节点190个、靶点节点275个。共筛选出丹蒌片与DCM相关靶点19个,在DCM中具有作用的靶点相关通路103条。在体内试验中,丹蒌片干预减少DCM大鼠心肌组织中纤维化相关因子信使RNA的表达,抑制晚期糖基化终末产物受体(receptor for advanced glycation end products,RAGE)/核因子(nuclear factor,NF)-κB/核苷酸结合寡聚化结构域样受体蛋白(nucleotide-binding oligomerization domain-like receptor protein,NLRP)3通路相关蛋白RAGE、NLRP3的表达,以及NF-κB(p65)蛋白的磷酸化。结论丹蒌片通过阻断RAGE/NF-κB/NLRP3信号通路活化,抑制DCM大鼠心肌组织纤维化,进而抑制疾病进展。

Objective To systematically evaluate the pharmacological effects and molecular mechanisms of Danlou tablets with comprehensive pharmacological strategies,and to provide a theoretical basis for the treatment of diabetic cardiomyopathy(DCM).Methods The chemical constituents and targets of Danlou tablets were discovered from the platform of pharmacology analysis of traditional Chinese medicine system,and the interaction network of"componenttarget"was established.All the targets of DCM were screened through the database,the relationship diagram of"componentdisease target"was constructed.The core targets were screened,and the gene ontology analysis and the related pathway enrichment were performed on the core targets.The DCM rat model was treated with Danlou tablets to verify the predicted important signaling pathways.Results A total of 190 compound nodes and 275 target nodes were screened from 10 important formulations of Danlou tablet.A total of 19 targets related to DCM were screened from Danlou tablet,and 103 target related pathways had role in DCM.In vivo,Danlou tablet reduced the expression of fibrosis-related factor messenger RNA in myocardium of DCM rats and inhibited the expression of receptor for advanced glycation end products(RAGE)/nuclear factor(NF)-κB/nucleotide-binding oligomerization domain-like receptor protein(NLRP)3 pathway-related proteins RAGE and NLRP3,and phosphorylation of NF-κB(p65)protein.Conclusion Danlou tablets can inhibit myocardial fibrosis in DCM rats by blocking the activation of RAGE/NF-κB/NLRP3 signaling pathway,and then inhibiting the progression of the disease.