Wang Yangyang, Sun Chuying, Cao Yutang, Jiao Tingying, Wang Kanglong, Li Jiaqi, Zhang Mengjiao, Jiang Jie, Zhong Xianchun, Yu Shuwu, Xu Hualing, Wang Jiawen, Yi Tong, Tian Xiaoting, Zhu Haiyan, Zhou Haifeng, Huang Chenggang, Wu Tong, Guo Xiaozhen, Xie Cen
National Key Laboratory of Lead Druggability Research, Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai, 201203, China; State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
School of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
J Ethnopharmacol. 2025 Feb 10;338(Pt 1):119025. doi: 10.1016/j.jep.2024.119025. Epub 2024 Nov 1.
Ulcerative colitis (UC), a chronic inflammatory bowel disease, has become a significant public health challenge due to the limited effectiveness of available therapies. Huoxiang Zhengqi (HXZQ), a well-established traditional Chinese formula, shows potential in managing UC, as suggested by clinical and pharmacological studies. However, the active components and mechanisms responsible for its effects remain unclear.
This study aimed to identify the bioactive components of HXZQ responsible for its therapeutic effects on UC and to elucidate their underlying mechanisms.
The effect of HXZQ against dextran sodium sulfate (DSS)-induced colitis was investigated. Ingredients in HXZQ were characterized and analyzed in colitic mice using liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS). In vitro, biological activity of compounds was assessed using lipopolysaccharide (LPS)-induced Ana-1 cells and bone marrow-derived macrophages (BMDMs), tumor necrosis factor-alpha (TNF-α)-induced Caco-2 cells, and isolated intestinal crypts from colitic mice. These results were confirmed in vivo. The targets of the components were identified through bioinformatics analysis and validated via molecular docking, enzyme inhibition assays, and in vivo experiments. Hematoxylin and eosin (HE) staining, periodic acid-Schiff (PAS) staining, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), western blotting, and quantitative real-time polymerase chain reaction (qPCR) were employed to confirm the pharmaceutical effects.
A clinical equivalent dose of HXZQ (2.5 mL/kg) effectively treated DSS-induced colitis. A total of 113 compounds were identified in HXZQ, with 35 compounds detected in colitic mice. Glycyrrhizic acid (GA) and patchouli alcohol (PA) emerged as key contributors to the anti-colitic effects of HXZQ. Further investigation revealed that HXZQ and its active components decreased the levels of pro-inflammatory cytokines TNF-α, interleukin-1β (IL-1β), and interleukin-6 (IL-6) in colon, likely by inhibiting nuclear factor kappa-B (NF-κB) signaling pathway. This inhibition indirectly activated the intestinal farnesoid X receptor (FXR) signaling pathway, correcting bile acid imbalances caused by colitis. Additionally, these components significantly enhanced the expression of tight junction proteins ZO-1 and Occludin, as well as the adhesion protein E-cadherin, and reduced goblet cell loss, thereby repairing intestinal barrier injury. Mechanistically, GA and PA were found to inhibit 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) activity, leading to increased local active corticosterone levels in the intestine to exert anti-inflammatory effects. Notably, the inhibition of 11β-HSD1 with the selective inhibitor BVT2733 (BVT) ameliorated colitis in mice.
HXZQ exhibits therapeutic effects on UC, primarily through GA and PA inhibiting 11β-HSD1. This suggests new natural therapy approaches for UC and positions 11β-HSD1 as a potential target for colitis treatment.
溃疡性结肠炎（UC）是一种慢性炎症性肠病，由于现有治疗方法效果有限，已成为一项重大的公共卫生挑战。临床和药理学研究表明，藿香正气（HXZQ）这一成熟的传统中药配方在治疗UC方面具有潜力。然而，其发挥作用的活性成分和机制仍不清楚。
本研究旨在确定HXZQ对UC具有治疗作用的生物活性成分，并阐明其潜在机制。
研究了HXZQ对葡聚糖硫酸钠（DSS）诱导的结肠炎的作用。使用液相色谱 - 质谱联用（LC-MS）和气相色谱 - 质谱联用（GC-MS）对结肠炎小鼠体内的HXZQ成分进行表征和分析。在体外，使用脂多糖（LPS）诱导的Ana-1细胞和骨髓来源的巨噬细胞（BMDM）、肿瘤坏死因子-α（TNF-α）诱导的Caco-2细胞以及来自结肠炎小鼠的分离肠隐窝评估化合物的生物活性。这些结果在体内得到证实。通过生物信息学分析确定成分的靶点，并通过分子对接、酶抑制试验和体内实验进行验证。采用苏木精和伊红（HE）染色、过碘酸希夫（PAS）染色、免疫组织化学、酶联免疫吸附测定（ELISA）、蛋白质印迹法和定量实时聚合酶链反应（qPCR）来确认药物效果。
临床等效剂量的HXZQ（2.5 mL/kg）有效治疗了DSS诱导的结肠炎。在HXZQ中总共鉴定出113种化合物，在结肠炎小鼠中检测到35种化合物。甘草酸（GA）和广藿香醇（PA）是HXZQ抗结肠炎作用的关键贡献者。进一步研究表明，HXZQ及其活性成分可能通过抑制核因子κB（NF-κB）信号通路降低结肠中促炎细胞因子肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的水平。这种抑制间接激活肠道法尼酯X受体（FXR）信号通路，纠正结肠炎引起的胆汁酸失衡。此外，这些成分显著增强紧密连接蛋白ZO-1和闭合蛋白以及黏附蛋白E-钙黏蛋白的表达，并减少杯状细胞损失，从而修复肠道屏障损伤。从机制上讲，发现GA和PA抑制11β-羟基类固醇脱氢酶1（11β-HSD1）的活性，导致肠道局部活性皮质酮水平升高以发挥抗炎作用。值得注意的是，用选择性抑制剂BVT2733（BVT）抑制11β-HSD1可改善小鼠结肠炎。
HXZQ对UC具有治疗作用，主要是通过GA和PA抑制1β-HSD1。这为UC提出了新的天然治疗方法，并将11β-HSD1定位为结肠炎治疗的潜在靶点。
