Miao Jie, Wei Cong, Wang Hong-Lian, Li Yu-Qing, Yu Xin-Ming, Yang Xiu, Su Hong-Wei, Li Ping, Wang Li
College of Integrated Chinese and Western Medicine, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou, China.
The Clinical Laboratory of the Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou, China.
Heliyon. 2024 Aug 8;10(16):e35728. doi: 10.1016/j.heliyon.2024.e35728. eCollection 2024 Aug 30.
Renal fibrosis represents a pivotal characteristic of chronic kidney disease (CKD), for which effective interventions are currently lacking. The Src kinase activates the phosphatidylinositol-3 kinases (PI3K)/Akt1 pathway to promote renal fibrosis, casting a promising target for anti-fibrosis treatment. Chaihuang-Yishen formula (CHYS), a traditional Chinese medicinal prescription, has a validated efficacy in the treatment of CKD, however, with the underlying mechanism unresolved. This study aimed to uncover the pharmacological mechanisms mediating the effect of CHYS in treating renal fibrosis using network pharmacology followed by experimental validation. The chemical compounds of CHYS were retrieved from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database or published literature, followed by the prediction of their targets using SwissTargetPrediction software. Disease (CKD/renal fibrosis)-related targets were retrieved from the Genecards database. Protein-protein interaction (PPI) network was generated using the drug-disease common targets and visualized in Cytoscape software. The drug-disease targets were further subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses by Metascape software. Additionally, the compound-target-pathway network was established in Cytosape to identify key compounds, targets, and pathways. Network pharmacology analysis screened out 96 active compounds and 837 potential targets within the 7 herbal/animal medicines of CHYS, among which 237 drug-disease common targets were identified. GO and KEGG analysis revealed the enrichment of fibrosis-related biological processes and pathways among the 237 common targets. Compound-target-pathway network analysis highlighted protein kinases Src and Akt1 as the top two targets associated with the anti-renal fibrosis effects of CHYS. In UUO mice, treatment with CHYS attenuates renal fibrosis, accompanied by suppressed expression and phosphorylation activation of Src. Unlike Src, CHYS reduced Akt1 phosphorylation without affecting its expression. In summary, network pharmacology and in vivo evidence suggest that CHYS exerts its anti-renal fibrosis effects, at least in part, by inhibiting the Src/Akt1 signaling axis.
肾纤维化是慢性肾脏病（CKD）的一个关键特征，目前缺乏有效的干预措施。Src激酶激活磷脂酰肌醇-3激酶（PI3K）/Akt1通路以促进肾纤维化，这使其成为抗纤维化治疗的一个有前景的靶点。柴黄益肾方（CHYS）是一种中药方剂，在治疗CKD方面具有已证实的疗效，但其潜在机制尚未明确。本研究旨在利用网络药理学揭示CHYS治疗肾纤维化作用的药理机制，并进行实验验证。从中药系统药理学（TCMSP）数据库或已发表文献中检索CHYS的化学成分，然后使用SwissTargetPrediction软件预测其靶点。从Genecards数据库中检索疾病（CKD/肾纤维化）相关靶点。利用药物-疾病共同靶点生成蛋白质-蛋白质相互作用（PPI）网络，并在Cytoscape软件中进行可视化。通过Metascape软件对药物-疾病靶点进一步进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。此外，在Cytosape中建立化合物-靶点-通路网络以识别关键化合物、靶点和通路。网络药理学分析在CHYS的7种草药/动物药中筛选出96种活性化合物和837个潜在靶点，其中鉴定出237个药物-疾病共同靶点。GO和KEGG分析揭示了237个共同靶点中与纤维化相关的生物学过程和通路的富集。化合物-靶点-通路网络分析突出显示蛋白激酶Src和Akt1是与CHYS抗肾纤维化作用相关的前两个靶点。在单侧输尿管梗阻（UUO）小鼠中，CHYS治疗可减轻肾纤维化，同时伴有Src表达和磷酸化激活的抑制。与Src不同，CHYS可降低Akt1磷酸化水平，但不影响其表达。总之，网络药理学和体内证据表明，CHYS至少部分通过抑制Src/Akt1信号轴发挥其抗肾纤维化作用。
