目的 探明三黄泻心汤（Sanhuang Xiexin Decoction，SXD）中存在自组装纳米粒，并分析其主要有效成分，进一步选择小檗碱、黄芩苷和鞣酸3个有效成分自组装形成纳米粒，同时探究该纳米粒自组装机制及其对胃溃疡的治疗作用。方法 首先采用高速低温离心分离出SXD合煎液中的自组装纳米粒，通过激光粒度仪、场发射透射电子显微镜（field emission transmission electron microscope，FETEM）等方法对其形貌进行表征，验证复方合煎液中自组装纳米粒的存在，再利用超高效液相色谱-四级杆-飞行时间串联质谱（UPLC-quadrupole-time of flight mass spectrometry，UPLC-Q-TOF-MS）技术，对SXD中自组装纳米粒进行成分分析。选择小檗碱、黄芩苷、鞣酸3个有效成分，制备成小檗碱-黄芩苷-鞣酸自组装纳米粒（berberine-baicalin-tannic acid self-assembled nanoparticles，BBT@SAN），采用TEM、X射线单晶衍射（X-ray single crystal diffraction，XRD）、差示扫描量热（differential scanning calorimetry，DSC）分析等方法，对其开展表征研究。采用分子模拟对接技术推测分子结合位点，进一步通过¹H-NMR验证自组装机制。然后进行抗溃疡作用评价，建立乙醇胃溃疡模型，对黏膜损伤程度进行评分；采用苏木精-伊红（hematoxylin-eosin，HE）染色和过碘酸-雪夫（periodic acid-Schiff，PAS）染色观察小鼠胃黏膜损伤情况；使用ELISA试剂盒测定小鼠血清中的肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-1β、IL-10、谷胱甘肽（glutathione，GSH）、超氧化物歧化酶（superoxide dismutase，SOD）、过氧化氢酶（catalase，CAT）含量，探究BBT@SAN抗炎抗氧化活性。结果 粒径、ζ电位、TEM等结果表明SXD中存在自组装纳米粒，且分布均匀稳定，其中主要含有酸性的黄酮类和碱性的生物碱类有效成分，以小檗碱和黄芩苷为代表。基于SXD中3种自组装有效成分的BBT@SAN呈球形，且表面饱满光滑。XRD、DSC、分子模拟对接及¹H-NMR等结果表明，SXD中3种有效成分之间主要通过氢键和共轭作用发生结合，其中小檗碱的异喹啉环、黄芩苷的羧基和羟基以及鞣酸的葡萄糖环和酚羟基等参与了自组装形成。抗溃疡作用结果表明，BBT@SAN能够显著降低胃黏膜病理评分，减轻小鼠胃黏膜剥离程度，减少了炎症细胞浸润，调节胃溃疡小鼠血清中的炎症因子TNF-α、IL-1β、IL-6、IL-10以及氧化应激指标GSH、SOD、CAT水平，发挥抗炎抗氧化应激作用，减轻胃黏膜损伤。结论 SXD中主要有效成分小檗碱、黄芩苷和鞣酸自组装形成的纳米粒BBT@SAN，能够减轻小鼠胃黏膜损伤，具有抗炎、抗氧化作用。有利于从汤液自组装角度揭示SXD制剂特征与药效的相关性，为传统中药汤剂药效物质基础形态与药效的关联性提供一定依据。

Objective This study aims to elucidate the presence of self-assembled nanoparticles in Sanhuang Xiexin Decoction (SXD, 三黄泻心汤), and analyze the main active ingredients. Further, berberine, baicalin, and tannic acid were selected as the three active ingredients to self assemble into nanoparticles, and the self-assembly mechanism of the nanoparticles and their therapeutic effect on gastric ulcers were explored. Methods Firstly, self-assembled nanoparticles in SXD decoction were separated by high-speed and low-temperature centrifugation, and then analyzed using laser particle size analyzer and field emission transmission electron microscope (FETEM), characterization of its morphology using methods such as microscope and transmission electron microscopy (TEM) was performed to verify the presence of self-assembled nanoparticles in the compound decoction, followed by the use of UPLC-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS) perform compositional analysis of self-assembled nanoparticles in SXD using the UPLC-Q-TOF-MS technique. The self-assembled nanoparticles of berberine, baicalin and tannic acid, the main active ingredient in SXD, was characterized using TEM, X-ray single crystal diffraction (XRD), differential scanning calorimetry (DSC), and other methods. Using molecular simulation docking technology to speculate molecular binding sites, and further verifying the self-assembly mechanism through ¹H-NMR. Then, a preliminary evaluation of the anti-ulcer effect was conducted, and an ethanol gastric ulcer model was established. After euthanizing the mice, the intact stomach was exposed, and the gastric mucosal surface was unfolded and photographed to score the damage. hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining were used to observe the gastric mucosal damage in mice; To investigate the anti-gastric ulcer activity of BBT@SAN, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, IL-10, glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) in mouse serum was measured using ELISA kits. Results The results of particle size, ζ potential, TEM, etc. indicate the presence of self-assembled nanoparticles in SXD, which are uniformly and stably distributed. They mainly contain acidic flavonoids and alkaline alkaloids, represented by berberine and baicalin. It is spherical in shape and has a full and smooth surface. The three active ingredients in SXD mainly bind through hydrogen bonding and conjugation, among which the isoquinoline ring of berberine, the carboxyl and hydroxyl groups of baicalin, and the glucose and phenolic hydroxyl groups of tannic acid participate in the self-assembly process. Preliminary anti-gastric ulcer results indicate that BBT@SAN can significantly reduce the pathological score of gastric mucosa and regulate the levels of TNF-α, IL-1β, IL-6, IL-10, GSH, SOD and CAT in the serum of mice with gastric ulcers, improve inflammation and oxidative stress response in mice, and alleviate gastric mucosal damage. Conclusion The main active ingredients in SXD, berberine, baicalin, and tannic acid, can efficiently self-assemble into nanoparticles, which can alleviate gastric mucosal damage in mice and have anti-inflammatory and antioxidant effects. This study helps reveal the correlation between the formulation characteristics and pharmacological effects of SXD from the perspective of self-assembled herbal decoctions, providing evidence for the relationship between the morphological forms of active substances in traditional Chinese herbal decoctions and their therapeutic efficacy.

R283.6

国家自然科学基金-区域创新发展联合基金重点项目（U21A201100）；四川省自然科学基金（2023NSFSC0688）；四川省自然科学基金（2025ZNSFSC0599）；四川省中医药管理局项目（2023MS532）；成都中医药大学“杏林学者”青基人才专项（QJRC2023017）