Liu Qinwen, Li Xiaowei, Li Yi, Luo Qian, Fan Qiling, Lu Aiping, Guan Daogang, Li Jiahui
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Single Cell Technology and Application, Southern Medical University, Guangzhou, China.
Chin Med. 2024 Mar 1;19(1):36. doi: 10.1186/s13020-024-00896-z.
Liver cirrhosis is a chronic liver disease with hepatocyte necrosis and lesion. As one of the TCM formulas Wuling Powder (WLP) is widely used in the treatment of liver cirrhosis. However, it's key functional components and action mechanism still remain unclear. We attempted to explore the Key Group of Effective Components (KGEC) of WLP in the treatment of Liver cirrhosis through integrative pharmacology combined with experiments.
The components and potential target genes of WLP were extracted from published databases. A novel node importance calculation model considering both node control force and node bridging force is designed to construct the Function Response Space (FRS) and obtain key effector proteins. The genetic knapsack algorithm was employed to select KGEC. The effectiveness and reliability of KGEC were evaluated at the functional level by using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Finally, the effectiveness and potential mechanism of KGEC were confirmed by CCK-8, qPCR and Western blot.
940 effective proteins were obtained in FRS. KEGG pathways and GO terms enrichments analysis suggested that effective proteins well reflect liver cirrhosis characteristics at the functional level. 29 components of WLP were defined as KGEC, which covered 100% of the targets of the effective proteins. Additionally, the pathways enriched for the KGEC targets accounted for 83.33% of the shared genes between the targets and the pathogenic genes enrichment pathways. Three components scopoletin, caryophyllene oxide, and hydroxyzinamic acid from KGEC were selected for in vivo verification. The qPCR results demonstrated that all three components significantly reduced the mRNA levels of COL1A1 in TGF-β1-induced liver cirrhosis model. Furthermore, the Western blot assay indicated that these components acted synergistically to target the NF-κB, AMPK/p38, cAMP, and PI3K/AKT pathways, thus inhibiting the progression of liver cirrhosis.
In summary, we have developed a new model that reveals the key components and potential mechanisms of WLP for the treatment of liver cirrhosis. This model provides a reference for the secondary development of WLP and offers a methodological strategy for studying TCM formulas.
肝硬化是一种伴有肝细胞坏死和损伤的慢性肝病。五苓散（WLP）作为中医方剂之一，被广泛用于治疗肝硬化。然而，其关键功能成分及作用机制仍不清楚。我们试图通过整合药理学结合实验来探索WLP治疗肝硬化的关键有效成分组（KGEC）。
从已发表的数据库中提取WLP的成分和潜在靶基因。设计一种同时考虑节点控制力和节点桥接力的新型节点重要性计算模型，以构建功能反应空间（FRS）并获得关键效应蛋白。采用遗传背包算法选择KGEC。通过基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析，在功能水平上评估KGEC的有效性和可靠性。最后，通过CCK - 8、qPCR和蛋白质印迹法确认KGEC的有效性和潜在机制。
在FRS中获得了940个有效蛋白。KEGG通路和GO术语富集分析表明，有效蛋白在功能水平上很好地反映了肝硬化的特征。WLP的29种成分被定义为KGEC，其覆盖了有效蛋白靶点的100%。此外，KGEC靶点富集的通路占靶点与致病基因富集通路之间共享基因的83.33%。从KGEC中选择了东莨菪素、氧化石竹烯和羟基肉桂酸三种成分进行体内验证。qPCR结果表明，这三种成分均显著降低了转化生长因子 - β1诱导的肝硬化模型中COL1A1的mRNA水平。此外，蛋白质印迹分析表明，这些成分协同作用于NF - κB、AMPK/p38、cAMP和PI3K/AKT通路，从而抑制肝硬化的进展。
总之，我们开发了一种新模型，揭示了WLP治疗肝硬化的关键成分和潜在机制。该模型为WLP的二次开发提供了参考，并为研究中药方剂提供了一种方法策略。
