Zhang Hui, Xiong Zhekun, He Yanshan, Su Huixia, Jiao Yali
Department of Gastroenterology, Zhongshan Hospital of Traditional Chinese Medicine, Zhongshan, 528401, Guangdong, China.
Xinzhuang Community Health Center, No. 115 Xinjian Road, Minhang District, Shanghai, 201199, China.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Mar;398(3):2897-2908. doi: 10.1007/s00210-024-03433-9. Epub 2024 Sep 20.
Irritable bowel syndrome (IBS) is a prevalent gastrointestinal dysfunction. Cimifugin is an active component of Radix saposhnikoviae which is effective for maintaining intestinal barrier integrity and intestinal function. This study aimed to investigate the treatment efficacy of Cimifugin on intestinal barrier dysfunction and to unveil the relevant mechanism through network pharmacology and experimental verification as well as molecular docking. Through SuperPred and Pubchem databases, the targets of Cimifugin were obtained. The disease targets were screened using Disgenet and GEO databases. With STRING database and Cytoscape software, the analysis of PPI network was performed. In DAVID database, the hub genes of Cimifugin were analyzed using GO and Pathway enrichment analyses. To validate the binding of Cimifugin with core targets, molecular docking was performed. The in vitro cellular model of intestinal barrier was established via the induction of Caco2 cells with LPS. TEER was used to detect epithelial barrier function and permeability was measured using FITC-dextran (FD4). Western blotting was used to measure the expressions of SIRT1, tight junction proteins, and NRF2/HO-1 signaling pathway-related proteins. The fluorescence intensity of ZO-1, Occludin, and Claudin-1 was detected using immunofluorescence staining. ELISA was used to detect the expression levels of inflammatory cytokines. Through the integration of all targets of IBS and Cimifugin, 94 frequent drug-disease-related targets were identified. These targets were enriched in some signaling pathways, like cellular responses to stress, cellular responses to stimuli, and VEGFA-VEGFR2. Ten hub genes including PTGS2, ANPRP, TGFB1, ACACA, SIRT1, NEF2L2, APEX1, IL6, AKT1, and HSP90AB1 were obtained. Cimifugin showed strong affinity with four key genes, including AKT1, SIRT1, IL6, and NFE2L2 (NRF2), which were obtained through the intersection of hug genes with cellular responses to stimuli. In vitro experiments showed that Cimifugin ameliorated LPS-induced intestinal barrier injury in Caco2 cells via upregulating SIRT1 to modulate NRF2/HO-1 signaling pathway. Cimifugin could alleviate intestinal barrier dysfunction in IBS by upregulating SIRT1 to regulate the NRF2/HO-1 signaling pathway.
肠易激综合征（IBS）是一种常见的胃肠功能障碍。升麻素是防风的一种活性成分，对维持肠道屏障完整性和肠道功能有效。本研究旨在探讨升麻素对肠道屏障功能障碍的治疗效果，并通过网络药理学、实验验证以及分子对接揭示其相关机制。通过SuperPred和Pubchem数据库获取升麻素的靶点。利用Disgenet和GEO数据库筛选疾病靶点。借助STRING数据库和Cytoscape软件进行蛋白质-蛋白质相互作用（PPI）网络分析。在DAVID数据库中，使用基因本体（GO）和通路富集分析对升麻素的枢纽基因进行分析。为验证升麻素与核心靶点的结合，进行分子对接。通过用脂多糖（LPS）诱导Caco2细胞建立肠道屏障的体外细胞模型。采用跨上皮电阻（TEER）检测上皮屏障功能，使用异硫氰酸荧光素标记的葡聚糖（FD4）测量通透性。采用蛋白质免疫印迹法检测沉默信息调节因子1（SIRT1）、紧密连接蛋白以及核因子E2相关因子2/血红素加氧酶-1（NRF2/HO-1）信号通路相关蛋白的表达。使用免疫荧光染色检测闭合蛋白1（ZO-1）、闭合蛋白（Occludin）和Claudin-1的荧光强度。采用酶联免疫吸附测定（ELISA）检测炎性细胞因子的表达水平。通过整合IBS和升麻素的所有靶点，确定了94个常见的药物-疾病相关靶点。这些靶点富集于一些信号通路，如细胞对应激的反应、细胞对刺激的反应以及血管内皮生长因子A（VEGFA）-血管内皮生长因子受体2（VEGFR2）。获得了包括前列腺素内过氧化物合酶2（PTGS2）、血管紧张素原（ANPRP）、转化生长因子β1（TGFB1）、乙酰辅酶A羧化酶α（ACACA）、SIRT1、核因子E2相关因子2（NEF2L2）、脱嘌呤/脱嘧啶核酸内切酶1（APEX1）、白细胞介素6（IL6）、蛋白激酶B（AKT1）和热休克蛋白90α家族成员1（HSP90AB1）在内的10个枢纽基因。升麻素与通过枢纽基因与细胞对刺激的反应的交集获得的4个关键基因，即AKT1、SIRT1、IL6和核因子E2相关因子2（NRF2）具有很强的亲和力。体外实验表明，升麻素通过上调SIRT1来调节NRF2/HO-1信号通路，改善LPS诱导的Caco2细胞肠道屏障损伤。升麻素可通过上调SIRT1来调节NRF2/HO-1信号通路，缓解IBS中的肠道屏障功能障碍。
