基于生物信息学和体外实验验证的肾康注射液治疗肾纤维化作用机制研究

Mechanism of Shenkang injection in treatment of renal fibrosis based on bioinformatics and in vitro experimental verification

目的 利用生物信息学及体外实验探讨肾康注射液(Shenkang injection, SKI)治疗肾纤维化(renal fibrosis, RF)的作用机制与物质基础。方法 利用GEO数据库筛选RF差异表达基因,借助CMAP数据库,基于基因表达谱相似性原理,重定位出具有调控RF作用的药物,然后通过分子指纹相似性分析筛选出SKI潜在治疗RF的成分;同时基于网络药理学预测SKI调控RF的核心靶点及通路;最后,通过分子对接和细胞实验进行验证。结果 基于GEO数据库筛选到2个RF相关的数据集,CMAP重定位到3个共同的RF治疗药物(saracatinib、dasatinib、PP-2),分子指纹相似性分析发现,RF治疗药物与salvianolic acid B、hydroxysafflor yellow A等5个SKI成分的结构相似性较高。分子对接结果发现,salvianolic acid B、hydroxysafflor yellow A等成分与SKI调控的潜在治疗RF的核心靶标MMP1、MMP13等均有良好的结合能力。网络药理学分析提示,SKI核心靶点主要富集在Relaxin和AGE-RAGE等信号通路。细胞实验表明,SKI可显著降低RF模型细胞AGE-RAGE信号通路中AGER、NFKB1、COL1A1、SERPINE1、VEGFC和Relaxin信号通路中MMP1、MMP13的mRNA表达水平,显著升高RXFP1的mRNA表达水平。结论 SKI可通过调控Relaxin和AGE-RAGE信号通路来发挥治疗RF的作用,其物质基础可能为salvianolic acid B、hydroxysafflor yellow A等成分。

Aim To explore the mechanism and material basis of Shenkang injection(SKI)in the treatment of renal fibrosis(RF)by bioinformatics and in vitro experiments.Methods The differentially expressed genes of RF were screened by GEO database.With the help of CMAP database,based on the similarity principle of gene expression profile,the drugs that regulated RF were repositioned,and then the components of SKI potential treatment RF were screened by molecular fingerprint similarity analysis.At the same time,the core targets and pathways of SKI regulating RF were predicted based on network pharmacology.Finally,it was verified by molecular docking and cell experiments.Results Based on the GEO database,two RF-related data sets were screened,and CMAP was relocated to three common RF therapeutic drugs(saracatinib,dasatinib,pp-2).Molecular fingerprint similarity analysis showed that RF therapeutic drugs had high structural similarity with five SKI components such as salvianolic acid B and hydroxysafflor yellow A.Molecular docking results showed that salvianolic acid B,hydroxysafflor yellow A and other components had good binding ability with MMP1 and MMP13,which were the core targets of SKI-regulated potential treatment of RF.Network pharmacology analysis suggested that the core targets of SKI were mainly enriched in signaling pathways such as Relaxin and AGE-RAGE.Cell experiments showed that SKI could significantly reduce the mRNA expression levels of AGER,NFKB1,COL1A1,SERPINE1,VEGFC in AGE-RAGE signaling pathway and MMP1 and MMP13 in Relaxin signaling pathway in RF model cells,and significantly increase the mRNA expression level of RXFP1.Conclusions SKI can play a role in the treatment of RF by regulating Relaxin and AGE-RAGE signaling pathways,and its material basis may be salvianolic acid B,hydroxysafflor yellow A and other components.