Liu Xin, Gong Qianqian, Deng Xianglan, Li Longxuan, Luo Ruiyi, Li Xuemin, Guo Dale, Deng Fang
State Key Laboratory of Southwestern Chinese Medicine Resources, The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.
State Key Laboratory of Southwestern Chinese Medicine Resources, The Ministry of Education Key Laboratory of Standardization of Chinese Herbal Medicine, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 2):118926. doi: 10.1016/j.jep.2024.118926. Epub 2024 Oct 10.
ETHNOPHARMACOLOGICAL RELEVANCE: The prevalence of gastrointestinal motility disorders (GMD) is increasing and is characterized by long-term recurrence. Citri Sarcodactylis Fructus (CSF), as a traditional Chinese medicine (TCM) known in "regulating qi and harmonizing the stomach", has therapeutic effects on GMD. However, the material basis of its efficacy is not clear. AIM OF THE STUDY: The aim of this study was to evaluate the gastrointestinal motility-promoting activity of CSF extracts and to screen their active ingredients and to perform a preliminary validation. METHODS: The chemical composition spectrum of different extracts of CSF were established by ultra high-performance liquid chromatography coupled with quadrupole orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS). The gastrointestinal motility-promoting activities of CSF were investigated by determining the intestinal propulsion rate, gastric emptying rate, acetylcholinesterase activity, and motilin content in L-arginine-induced GMD mice. Spectrum-effect relationship and network pharmacology analysis were used for the screening of potential active ingredients. A zebrafish gastrointestinal motility model traced with Nile Red was established to validate the active ingredients. Molecular docking prediction was used to explore the mechanism of action of the active ingredient. Finally, Western blotting and TUNEL staining were performed to validate the molecular docking predictions. RESULTS: In total, 42 shared components were identified. The main active fraction of CSF to promote gastrointestinal motility was 70% ethanol elution fraction. Eleven potential active ingredients were screened by grey correlation analysis, orthogonal partial least squares analysis, and "active ingredient-target" network. Six compounds were confirmed as the pharmacodynamic substances of CSF by zebrafish gastrointestinal motility model, namely, quercetin, kaempferol, isorhamnetin, diosmetin, hesperetin, and 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone. Molecular docking predictions and Western blotting assays indicated that CSF may act on AKT and MMP9 targets to exert gastrointestinal motility-promoting activity. CONCLUSION: This study provided a foundation for elucidating the gastrointestinal motility-promoting activity of CSF and its material basis by integrating spectrum-effect relationship and network pharmacology. It also provided a theoretical basis for quality control of CSF and a new idea for the discovery and validation of pharmacodynamic substances in TCM.
民族药理学相关性：胃肠动力障碍（GMD）的患病率正在上升，且具有长期复发的特点。佛手（CSF）作为一种以“理气和胃”著称的传统中药，对GMD具有治疗作用。然而，其疗效的物质基础尚不清楚。 研究目的：本研究旨在评估CSF提取物促进胃肠动力的活性，筛选其活性成分并进行初步验证。 方法：采用超高效液相色谱-四极杆轨道阱高分辨质谱联用（UHPLC-Q/Orbitrap HRMS）建立CSF不同提取物的化学成分谱。通过测定L-精氨酸诱导的GMD小鼠的肠推进率、胃排空率、乙酰胆碱酯酶活性和胃动素含量，研究CSF促进胃肠动力的活性。采用谱效关系和网络药理学分析筛选潜在活性成分。建立尼罗红标记的斑马鱼胃肠动力模型验证活性成分。利用分子对接预测探索活性成分的作用机制。最后，进行蛋白质免疫印迹法（Western blotting）和末端脱氧核苷酸转移酶介导的缺口末端标记法（TUNEL）染色验证分子对接预测结果。 结果：共鉴定出42种共有成分。CSF促进胃肠动力的主要活性部位为70%乙醇洗脱部位。通过灰色关联分析、正交偏最小二乘法分析和“活性成分-靶点”网络筛选出11种潜在活性成分。通过斑马鱼胃肠动力模型确认6种化合物为CSF的药效物质，即槲皮素、山柰酚、异鼠李素、香叶木素、橙皮素和5,7,3'-三羟基-6,4',5'-三甲氧基黄酮。分子对接预测和蛋白质免疫印迹法检测表明，CSF可能作用于AKT和MMP9靶点发挥促进胃肠动力的活性。 结论：本研究通过整合谱效关系和网络药理学，为阐明CSF促进胃肠动力的活性及其物质基础提供了依据。也为CSF的质量控制提供了理论基础，为中药药效物质的发现和验证提供了新思路。
J Chromatogr B Analyt Technol Biomed Life Sci. 2024-2-1
Zhongguo Zhong Yao Za Zhi. 2024-7
Zotero 插件