Wang Tao, Ou Yanghui, Xiong Wenjun, Luo Lijie, Xu Yihua, Chen Guojian, Peng Yaohui, Chen Yan, Zeng Haiping, Yu Yang, Tang Haipeng, Yao Hongliang, Wang Wei
Department of Gastrointestinal Surgery, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510405, People's Republic of China.
Guangdong Clinical Research Institute of Chinese Medicine, Guangzhou, Guangdong, 510405, People's Republic of China.
J Inflamm Res. 2025 Jun 7;18:7301-7321. doi: 10.2147/JIR.S519483. eCollection 2025.
Colitis, a inflammatory disorder of the colon, causes significant morbidity and declines quality of life. Despite treatment advancements, effective therapies remain limited. Wuda Granules (WDG), a traditional medicine formula, is clinically used for intestinal obstruction and gastrointestinal recovery through its anti-inflammatory effects.
Network pharmacology and molecular docking were employed to identify the active compounds, key targets, and associated signaling pathways of WDG against Colitis. A mouse model of DSS-induced colitis was established to evaluate the therapeutic potential of WDG. Colitis-related symptoms, including weight loss, DAI score, spleen index, and colon length, were measured. Histopathological changes were analyzed using H&E staining. To assess intestinal barrier integrity, goblet cell abundance was determined by AB-PAS staining, and the expression of related proteins was analyzed by IHC. Inflammatory and oxidative stress markers were measured by ELISA, biochemical assays, and Western blotting.
Network pharmacology analysis identified the core therapeutic targets of WDG against UC is IL-6, TP53, AKT1, IL-1β, and TNF. WDG treatment significantly improved colitis-related symptoms, as evidenced by reduced weight loss, DAI score, and spleen index, as well as increased colon length. Histological analysis revealed preserved colon structure and reduced inflammatory infiltration. WDG suppressed the expression of pro-inflammatory mediators (IL-6, IL-1β, TNF-α, MPO) and enhanced the levels of anti-inflammatory cytokines (IL-10 and IL-22). In addition, WDG alleviated lung inflammation and inhibited M1 macrophage polarization. Intestinal barrier integrity was improved by increasing goblet cell numbers and upregulating the expression of MUC2, ZO-1, Occludin, Claudin-1. Antioxidant activity was enhanced, indicated by elevated SOD and CAT levels and decreased MDA content. Mechanistically, Western blot analysis showed that WDG activated the Nrf2/Keap1/HO-1 signaling pathway. Molecular docking further revealed one potential active compound, 6,7-Dimethoxy-2-(2-phenylethyl) chromone, which exhibited strong binding affinity with Nrf2/Keap1 complex.
WDG alleviates DSS-induced colitis by inhibiting inflammation, enhancing intestinal barrier integrity, and reducing oxidative stress through activation of the Nrf2/Keap1/HO-1 pathway.
结肠炎是一种结肠炎症性疾病，会导致严重的发病率并降低生活质量。尽管治疗方法有所进步，但有效的治疗手段仍然有限。五答颗粒（WDG）是一种传统中药配方，因其抗炎作用在临床上用于治疗肠梗阻和促进胃肠道恢复。
采用网络药理学和分子对接技术来确定WDG抗结肠炎的活性成分、关键靶点和相关信号通路。建立葡聚糖硫酸钠（DSS）诱导的小鼠结肠炎模型以评估WDG的治疗潜力。测量与结肠炎相关的症状，包括体重减轻、疾病活动指数（DAI）评分、脾脏指数和结肠长度。使用苏木精-伊红（H&E）染色分析组织病理学变化。为评估肠道屏障完整性，通过阿尔新蓝-过碘酸雪夫（AB-PAS）染色确定杯状细胞丰度，并通过免疫组织化学（IHC）分析相关蛋白的表达。通过酶联免疫吸附测定（ELISA）、生化分析和蛋白质印迹法测量炎症和氧化应激标志物。
网络药理学分析确定WDG抗溃疡性结肠炎（UC）的核心治疗靶点为白细胞介素-6（IL-6）、肿瘤蛋白p53（TP53）、蛋白激酶B（AKT1）、白细胞介素-1β（IL-1β）和肿瘤坏死因子（TNF）。WDG治疗显著改善了与结肠炎相关的症状，体重减轻、DAI评分和脾脏指数降低以及结肠长度增加证明了这一点。组织学分析显示结肠结构得以保留且炎症浸润减少。WDG抑制促炎介质（IL-6、IL-1β、TNF-α、髓过氧化物酶（MPO））的表达并提高抗炎细胞因子（IL-10和IL-22）的水平。此外，WDG减轻肺部炎症并抑制M1巨噬细胞极化。通过增加杯状细胞数量和上调黏蛋白2（MUC2）、紧密连接蛋白1（ZO-1）、闭合蛋白（Occludin）、Claudin-1的表达改善肠道屏障完整性。超氧化物歧化酶（SOD）和过氧化氢酶（CAT）水平升高以及丙二醛（MDA）含量降低表明抗氧化活性增强。机制上，蛋白质印迹分析表明WDG激活了核因子E2相关因子2（Nrf2）/ Kelch样环氧氯丙烷相关蛋白1（Keap1）/血红素氧合酶-1（HO-1）信号通路。分子对接进一步揭示了一种潜在的活性化合物，6,7-二甲氧基-2-（2-苯乙基）色酮，它与Nrf2/Keap1复合物表现出很强的结合亲和力。
WDG通过抑制炎症、增强肠道屏障完整性以及通过激活Nrf2/Keap1/HO-1途径减轻氧化应激来缓解DSS诱导的结肠炎。
