Jiaen Huang, Qing Luo, Gengting Dong, Weiwen Peng, Jianhong H E, Weibo Dai
Department of Pharmacy, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine, Zhongshan 528400, China.
State Key Laboratory of Quality Research in Chinese Medicine & Faculty of Chinese Medicine, Macau University of Science and Technology, Macau SAR 999078, China.
J Tradit Chin Med. 2025 Apr;45(2):311-325. doi: 10.19852/j.cnki.jtcm.2025.02.006.
To evaluate the therapeutic effects of Xiahuo Pingwei San (, XHPWS) on ulcerative colitis (UC) in mice and to explore the underlying mechanisms through a network pharmacology approach.
Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was utilized to identify the chemical composition and authenticate the active constituents of XHPWS, ensuring rigorous quality control across batches. A dextran sulfate sodium (DSS)-induced UC model was established in C57BL/6 mice, which were treated with XHPWS . The efficacy against UC was assessed by measuring parameters such as body weight, disease activity index (DAI) scores, and colon length. Levels of inflammatory cytokines, including interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-alpha (TNF-α), in colonic tissue were evaluated using enzyme-linked immunosorbent assay (ELISA). Histological analysis of colon sections was conducted using hematoxylin and eosin staining. A network pharmacology approach was employed to explore the mechanisms of XHPWS and to predict its potential targets in UC treatment. Predicted protein expressions in colonic tissue were validated using immune-ohistochemistry (IHC) and Western blotting techniques.
XHPWS effectively alle ted DSS-induced UC symptoms in mice, as evidenced by restored body weight, reduced colon shortening, and decreased DAI scores. Histopathological examination revealed that XHPWS significantly reduced intestinal inflammatory infiltration, restored intestinal epithelial permeability, and increased goblet cell count. Network pharmacology analysis identified 63 active compounds in XHPWS and suggested that it might target 35 potential proteins associated with UC treatment. Functional enrichment analysis indicated that the protective mechanism of XHPWS could be related to the advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway. Notably, quercetin, kaempferol, wogonin, and nobiletin, the main components of XHPWS, showed strong correlations with the core targets. Additionally, experimental validation demonstrated that XHPWS significantly decreased levels of inflammatory cytokines interleukin 6 (IL-6), interleukin 1 beta (IL-1β), and tumor necrosis factor alpha (TNF-α) in UC mice, while downregulating the expression of proteins related to the AGE-RAGE pathway.
Our study demonstrated that XHPWS effectively alle tes colitis symptoms and inflammation in UC mice, potentially through the regulation of the AGE-RAGE pathway. These findings provide strong evidence for the therapeutic potential of XHPWS in UC treatment, thereby broadening its clinical applications.
评价泻火平胃散（XHPWS）对小鼠溃疡性结肠炎（UC）的治疗作用，并通过网络药理学方法探讨其潜在机制。
采用超高效液相色谱-四极杆飞行时间质谱联用技术（UPLC-Q-TOF/MS）鉴定XHPWS的化学成分并验证其活性成分，确保各批次质量控制严格。在C57BL/6小鼠中建立葡聚糖硫酸钠（DSS）诱导的UC模型，并用XHPWS进行治疗。通过测量体重、疾病活动指数（DAI）评分和结肠长度等参数评估对UC的疗效。使用酶联免疫吸附测定（ELISA）评估结肠组织中炎症细胞因子的水平，包括白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）。采用苏木精和伊红染色对结肠切片进行组织学分析。采用网络药理学方法探讨XHPWS的作用机制，并预测其在UC治疗中的潜在靶点。使用免疫组织化学（IHC）和蛋白质印迹技术验证结肠组织中预测的蛋白质表达。
XHPWS有效减轻了DSS诱导的小鼠UC症状，表现为体重恢复、结肠缩短减轻和DAI评分降低。组织病理学检查显示，XHPWS显著减少肠道炎症浸润，恢复肠道上皮通透性，并增加杯状细胞数量。网络药理学分析鉴定出XHPWS中的63种活性化合物，并表明其可能靶向35种与UC治疗相关的潜在蛋白质。功能富集分析表明，XHPWS的保护机制可能与晚期糖基化终末产物-晚期糖基化终末产物受体（AGE-RAGE）信号通路有关。值得注意的是，XHPWS的主要成分槲皮素、山奈酚、汉黄芩素和橙皮素与核心靶点显示出强相关性。此外，实验验证表明，XHPWS显著降低了UC小鼠中炎症细胞因子白细胞介素6（IL-6）、白细胞介素1β（IL-1β）和肿瘤坏死因子α（TNF-α）的水平，同时下调了与AGE-RAGE途径相关的蛋白质表达。
我们的研究表明，XHPWS可能通过调节AGE-RAGE途径有效减轻UC小鼠的结肠炎症状和炎症。这些发现为XHPWS在UC治疗中的治疗潜力提供了有力证据，从而拓宽了其临床应用。
