目的 采用微沉淀法制备大黄-牡丹皮新型自组装纳米粒（Rhei Radix et Rhizoma-MoutanCortex novel self-assembled nanoparticles，RMN-SAN）并开展表征、形成机制与抗结直肠癌（colorectal cancer，CRC）药效作用探究。方法 通过单因素实验优化工艺参数，开展动态光散射、ζ电位和透射电子显微镜等表征；利用UPLC-Q-TOF-MS技术鉴定核心活性成分，借助傅里叶变换红外光谱（Fourier transform infrared spectroscopy，FTIR）与UV光谱揭示RMN-SAN自组装机制；通过体外CT26细胞实验与体内荷瘤小鼠模型验证其抗CRC药效。结果 优化后RMN-SAN最佳制备方法为大黄、牡丹皮各10 g，加10倍量水，浸泡30 min，回流提取30 min，滤过。药渣加8倍70%乙醇回流提取30 min，滤过。合并2次提取液，600 r/min、25 ℃条件下磁力搅拌30 min，60 ℃旋转蒸发除醇，并浓缩至0.2 g/mL，梯度离心（4 ℃，6 000、8 000、10 000 r/min各10 min）后透析（截留相对分子质量3 500）12 h，即得RMN-SAN。RMN-SAN的平均粒径为（109.30±4.20）nm，PDI为0.353±0.009，ζ电位为（-24.5±1.3）mV，呈均一球形结构；共鉴定出大黄酸、芦荟大黄素、番泻苷B、丹皮酚和芍药苷5种核心成分，其自组装依赖氢键与π-π堆积作用驱动并得到了分子对接验证。体外实验证实RMN-SAN可浓度相关性抑制CT26细胞增殖、诱导凋亡和G₁期阻滞；体内实验中RMN-SAN高剂量（6.0 g/kg）组抑瘤作用显著高于传统煎煮法组，且能更高效地调节免疫炎症因子水平。结论 通过现代工艺赋能中药配伍优势，为中药自组装纳米粒的制备优化提供了新思路，也为CRC的中医药治疗提供了高效稳定的新型制剂。

Objective Novel self-assembled nanoparticles (RMN-SAN) of Dahuang (Rhei Radix et Rhizoma) and Mudanpi (Moutan Cortex) were prepared by the micro-precipitation method, and their characterization, formation mechanism and anti-colorectal cancer (CRC) efficacy were investigated. Methods The process parameters were optimized through single-factor experiments, and characterization methods such as dynamic light scattering, ζ potential and transmission electron microscopy were performed. The core active components were identified by UPLC-Q-TOF-MS, and the self-assembly mechanism of RMN-SAN was revealed by FTIR and UV spectroscopy. The in vitro CT26 cell experiments and in vivo tumor-bearing mouse models were used to verify its anti-CRC efficacy. Results The optimized preparation method for RMN-SAN involves taking 10 g each of Rhei Radix et Rhizoma and Moutan Cortex, adding 10 times their volume of water, soaking for 30 min, and refluxing for 0.5 h, followed by filtration. The residue is then refluxed with 8 times its volume of 70% ethanol for 0.5 h and filtered again. The two extracts are combined, magnetically stirred at 600 r/min and 25 ℃ for 30 min, and the alcohol is removed by rotary evaporation at 60 ℃, concentrating to a final concentration of 0.2 g/mL. The solution is subjected to gradient centrifugation (4 ℃, 6 000, 8 000, and 10 000 r/min, each for 10 min), followed by dialysis (cut-off molecular weight 3 500) for 12 h, yielding the final product. The results showed that the average particle size of the optimized RMN-SAN was (109.30 ± 4.20) nm, the PDI was 0.353 ± 0.009, and the ζ potential was (-24.5 ± 1.3) mV, presenting a uniform spherical structure. Five core components such as rhein, aloe-emodin, sennoside B, paeonol, and paeoniflorin were identified, and their self-assembly was driven by hydrogen bonds and π-π stacking interactions, which was verified by molecular docking. The in vitro experiments confirmed that RMN-SAN could inhibit the proliferation of CT26 cells in a concentration-dependent manner, induce apoptosis and block the cell cycle at the G₁ phase. In the in vivo experiments, the tumor suppression effect of the high-dose RMN-SAN group (6.0 g/kg) was significantly higher than that of the traditional decoction group, and it could regulate the levels of immune and inflammatory factors more efficiently. Conclusion This study enabled the utilization of the therapeutic advantages of traditional Chinese medicine through modern processing, providing new ideas for the optimization of the preparation of self-assembled nanoparticles of traditional Chinese medicine, and also providing an efficient and stable new formulation for the treatment of CRC with traditional Chinese medicine.

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2024年度博士后创新人才支持计划“A档”（原国家“博新计划”，BX20240048）；国家自然科学基金青年项目（82505368）；中国博士后科学基金第77批面上项目（2025M773871）；四川省科学技术厅青年项目（2024NSFSC1821）；成都市科技局技术创新研发项目（2026-YF05-01025-SN）；成都中医药大学2024年度“杏林学者”学科人才科研提升计划（BSZ2024033）