网络药理学分析结合实验验证探讨红景天苷对肠缺血再灌注的治疗机制。
Network pharmacology analysis combined with experimental validation to explore the therapeutic mechanism of salidroside on intestine ischemia reperfusion.
机构信息
The First Clinical Medical College of Lanzhou University, Lanzhou, 730000, GanSu Province, China.
Department of Anesthesiology, First Hospital of Lanzhou University, Lanzhou, 730000, GanSu Province, China.
出版信息
Biosci Rep. 2023 Aug 31;43(8). doi: 10.1042/BSR20230539.
ETHNOPHARMACOLOGICAL RELEVANCE
Salidroside (SAL), a phenolic natural product present in Rhodiola rosea, are commonly used in the treatment of various ischemic-hypoxic diseases, including intestinal ischemia-reperfusion (IR) injury. However, their efficacy and potential mechanisms in the treatment of intestinal IR injury have not been investigated.
OBJECTIVE
The objective of the present study is to investigate the pharmacological mechanism of action of SAL on intestinal IR injury using a network pharmacology approach combined with experimental validation.
METHODS
In the present study, we used the Traditional Chinese Medicine Systematic Pharmacology (TCMSP) database and analysis platform and Comparative Toxicogenomics Database (CTD) to predict possible target genes of SAL, collected relevant target genes of intestinal IR injury from GeneCards and DisGenet websites, and collected summary data to screen common target genes. Then, the protein-protein interaction (PPI) target network was constructed and analyzed by STRING database and Cytoscape 3.8.2 with the above intersecting genes. Then, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed and the component-target-pathway network was constructed, followed by the use of molecular docking and molecular dynamic simulation to verify the possible binding conformation between SAL and candidate targets to further explore the potential targets of SAL in the treatment of intestinal IR injury. Finally, an in vivo model of mouse superior mesenteric artery ligation was established to assess the anti-intestinal IR injury effect of SAL by assessing histopathological changes in mouse small intestine by HE staining, detecting inflammatory factor expression by ELISA kit, and detecting the expression of key protein targets by Western blotting.
RESULTS
A total of 166 SAL target genes and 1740 disease-related targets were retrieved, and 88 overlapping proteins were obtained as potential therapeutic targets. The pathway enrichment analysis revealed that the pharmacological effects of SAL on intestinal IR injury were anti-hypoxic, anti-inflammatory and metabolic pathway related, and the molecular docking and molecular dynamic simulation results showed that the core bioactive components had good binding affinity for TXNIP and AMPK, and the immunoblotting results indicated that the expression levels of TXNIP and AMPK in the small intestinal tissues of mice in the drug-treated group compared with the model group were significantly changed.
CONCLUSION
SAL may target AMPK and TXNIP domains to act as a therapeutic agent for intestinal IR. These findings comprehensively reveal the potential therapeutic targets for SAL against intestinal IR and provide theoretical basis for the clinical application of SAL in the treatment of intestinal IR.
民族药理学相关性
Salidroside（SAL）是一种存在于红景天中的酚类天然产物，常用于治疗各种缺血缺氧性疾病，包括肠缺血再灌注（IR）损伤。然而，其在治疗肠 IR 损伤方面的疗效和潜在机制尚未得到研究。
目的
本研究旨在采用网络药理学方法结合实验验证，探讨 SAL 对肠 IR 损伤的药理作用机制。
方法
本研究采用中药系统药理学（TCMSP）数据库和分析平台以及比较毒理学基因组学数据库（CTD）预测 SAL 的可能作用靶点，从 GeneCards 和 DisGenet 网站收集肠 IR 损伤的相关靶点，并进行汇总数据筛选共同靶点。然后，通过 STRING 数据库和 Cytoscape 3.8.2 构建和分析蛋白质-蛋白质相互作用（PPI）靶标网络。然后进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析，并构建成分-靶标-通路网络，再进行分子对接和分子动力学模拟，以验证 SAL 与候选靶标之间可能的结合构象，进一步探讨 SAL 治疗肠 IR 损伤的潜在靶标。最后，通过建立小鼠肠系膜上动脉结扎的体内模型，通过 HE 染色评估小鼠小肠的组织病理学变化、ELISA 试剂盒检测炎症因子表达、Western blot 检测关键蛋白靶标表达，评估 SAL 对肠 IR 损伤的抗作用。
结果
共检索到 166 个 SAL 作用靶点和 1740 个疾病相关靶点，获得 88 个重叠蛋白作为潜在治疗靶点。通路富集分析表明，SAL 对肠 IR 损伤的药理作用与抗缺氧、抗炎和代谢途径有关，分子对接和分子动力学模拟结果表明，核心生物活性成分与 TXNIP 和 AMPK 具有良好的结合亲和力，免疫印迹结果表明与模型组相比，药物治疗组小鼠小肠组织中 TXNIP 和 AMPK 的表达水平发生了明显变化。
结论
SAL 可能通过靶向 AMPK 和 TXNIP 结构域发挥作用，成为治疗肠 IR 的药物。这些发现全面揭示了 SAL 治疗肠 IR 的潜在治疗靶点，为 SAL 临床应用于肠 IR 治疗提供了理论依据。
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