Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Institute of Digestive Disease, Department of Medicine and Therapeutics, State Key Laboratory of Digestive Disease, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, Shenzhen Research Institute, Sha Tin, New Territories, Hong Kong, China.
J Ethnopharmacol. 2022 Aug 10;294:115324. doi: 10.1016/j.jep.2022.115324. Epub 2022 Apr 27.
Xiaoyaosan is a traditional Chinese herbal formula that has long been used to treat liver cirrhosis, liver failure, and hepatocarcinoma (HCC). However, little is known about its mechanism of action and targets in treating chronic liver disease.
This study aimed to detect the critical transition of HCC progression and to explore the regulatory mechanism and targets of Xiaoyaosan treating liver cirrhosis (cirrhosis) using integrative medicinal research involving system biology and pharmacology.
We recruited chronic liver disease participants to obtain gene expression data and applied the dynamic network biomarker (DNB) method to identify molecular markers and the critical transition. We combined network pharmacology and DNB analysis to locate the potential DNBs (targets). Then we validated the DNBs in the liver cirrhosis rat models using Xiaoyaosan treatment. The expression of genes encoding the four DNBs, including Cebpa, Csf1, Egfr, and Il7r, were further validated in rat liver tissue using Western blot analysis.
We found EGFR, CEBPA, Csf1, Ccnb1, Rrmm2, C3, Il7r, Ccna2, and Peg10 overlap in the DNB list and Xiaoyaosan-Target-Disease (XTD) network constructed using network pharmacology databases. We investigated the diagnostic ability of each member in the DNB cluster and found EGFR, CEBPA, CSF1, and IL7R had high diagnostic abilities with AUC >0.7 and P-value < 0.05. We validated these findings in rats and found that liver function improved significantly and fibrotic changes were relieved in the Xiaoyaosan treatment group. The expression levels of CSF1 and IL7R in the Xiaoyaosan group were significantly lower than those in the cirrhosis model group. In contrast, CEBPA expression in the Xiaoyaosan group was significantly higher than that in the cirrhosis model group. The expression of EGFR in the Xiaoyaosan group was slightly decreased than in the model group but not significantly.
Using the DNB method and network pharmacology approach, this study revealed that CEBPA, IL7R, EGFR, and CSF1 expression was remarkably altered in chronic liver disease and thus, may play an important role in driving the progression of cirrhosis. Therefore, CEBPA, IL7R, EGFR, and CSF1 may be important targets of Xiaoyaosan in treating cirrhosis and can be considered for developing novel therapeutics.
逍遥散是一种传统的中草药配方，长期以来一直用于治疗肝硬化、肝功能衰竭和肝癌（HCC）。然而，对于其在治疗慢性肝病中的作用机制和靶点知之甚少。
本研究旨在检测 HCC 进展的关键转变，并通过涉及系统生物学和药理学的综合医学研究，探索逍遥散治疗肝硬化（肝硬化）的调节机制和靶点。
我们招募了慢性肝病患者以获取基因表达数据，并应用动态网络生物标志物（DNB）方法来识别分子标志物和关键转变。我们结合网络药理学和 DNB 分析来定位潜在的 DNB（靶点）。然后，我们使用逍遥散治疗在肝硬化大鼠模型中验证了 DNB。使用 Western blot 分析进一步验证了基因编码的四个 DNB（包括 Cebpa、Csf1、Egfr 和 Il7r）在大鼠肝组织中的表达。
我们发现 EGFR、CEBPA、CSF1、CCNBl、RRMM2、C3、IL7R、CCNA2 和 PEG10 在 DNB 列表和使用网络药理学数据库构建的逍遥散-Target-Disease（XTD）网络中重叠。我们研究了 DNB 簇中每个成员的诊断能力，发现 EGFR、CEBPA、CSF1 和 IL7R 具有高诊断能力，AUC>0.7，P 值<0.05。我们在大鼠中验证了这些发现，并发现逍遥散治疗组肝功能明显改善，纤维化改变得到缓解。CSF1 和 IL7R 在逍遥散组的表达水平明显低于肝硬化模型组。相比之下，CEBPA 在逍遥散组的表达明显高于肝硬化模型组。EGFR 在逍遥散组的表达略低于模型组，但差异无统计学意义。
使用 DNB 方法和网络药理学方法，本研究表明，CEBPA、IL7R、EGFR 和 CSF1 的表达在慢性肝病中明显改变，因此可能在驱动肝硬化进展中起重要作用。因此，CEBPA、IL7R、EGFR 和 CSF1 可能是逍遥散治疗肝硬化的重要靶点，可以考虑开发新的治疗方法。
