目的 构建人源结直肠腺瘤（CRA）复发类器官，评价几种中药活性成分对CRA复发类器官活性的影响。方法 收集内镜下切除后再发腺瘤的CRA患者组织，腺瘤数量数目超过2枚且少于6枚，直径为5～10 mm，构建CRA复发三维（3D）类器官；使用高内涵智能分析仪和光学显微镜观察类器官形态；运用苏木精-伊红（HE）染色观察类器官组织学形态；采用外显子组测序检测CRA复发类器官与源组织基因学特征的一致性；采用CellTiter-Glo检测1 000.00、100.00、10.00、1.00、0.10、0.01 μmol·L^-1的盐酸小檗碱、人参皂苷Rg₃、大黄素、姜黄素、丹参酮Ⅱ A、黄芩素、异甘草素对6例CRA复发类器官活性的影响，绘制活性曲线，并计算半数抑制浓度（IC₅₀）值。结果 建成CRA复发类器官，形态多以内部透亮、近圆的囊样结构为主，且囊壁多有向外侧隆起的蕈状结构；经过多次传代、冻存再复苏后其形态结构均能保持一致；每组类器官与源组织之间的突变类型比例基本一致，插入和缺失突变的长度和比例、拷贝变异数均具有较高度相似性； 7种中药活性成分对6例CRA复发类器官的IC₅₀均值分别为盐酸小檗碱69.79 μmol·L^-1、人参皂苷Rg₃ 110.92 μmol·L^-1、大黄素14.33 μmol·L^-1、姜黄素4 352.02 μmol·L^-1、丹参酮Ⅱ A 1 772.97 μmol·L^-1、黄芩素16.01 μmol·L^-1、异甘草素78.48 μmol·L^-1，CRA复发类器官对大黄素和黄芩素具有更高的敏感性。结论 成功构建能够稳定传代、冻存和复苏的CRA复发类器官，其能维持源组织的组织学和基因学特征；经CRA复发类器官活性评价，大黄素和黄芩素具有治疗CRA复发的潜力，为中医药在治疗CRA复发和防治结直肠癌（CRC）领域的有效应用提供理论和方法学参考。

Objective To construct patient-derived colorectal adenoma (CRA) recurrence organoids, and to evaluate the effects of the ingredients of several traditional Chinese medicines on the activity of CRA recurrence organoids. Methods Tissues from CRA patients with recurrent adenomas after endoscopic resection were collected, and the number of adenomas was more than two and less than six, with diameters of 5—10 mm, to construct CRA recurrence 3D organoids; the morphology of the organoids was observed by using a high-content intelligent analyser and a light microscope; the histological morphology of the organoids was observed by using hematoxylin-eosin (HE) staining; whole-exome sequencing was used to test the consistency of the genetic characteristics of CRA recurrence organoids with those of the source tissues; CellTiter-Glo was used to test the effects of berberine, ginsenoside Rg₃, emodin, curcumin, tanshinone IIA, baicalein, and isoliquiritigenin of 1 000.00, 100.00, 10.00, 1.00, 0.10, 0.01 μmol·L^-1 on the activity of six CRA recurrence organoids, and the activity curves were plotted and the IC₅₀ values were calculated. Results The morphology of CRA recurrence organoids was mostly dominated by internal translucent, nearly rounded sac-like structures, and the sac walls were mostly myxoid with outward bulging; their morphology and structure could be maintained consistently after multiple passages, freezing and resuscitation; the proportions of mutation types were basically the same between the organoids and the source tissues of each group, and the lengths and proportions of insertion and deletion mutations as well as the number of copy variations had a high degree of similarity; the effects of the seven traditional Chinese medicines on the activity of six cases of CRA recurrence organoids were investigated. The mean IC₅₀ values of the seven active ingredients on the six CRA recurrence organoids were berberine 69.79 μmol·L^-1, ginsenoside Rg₃ 110.92 μmol·L^-1, emodin 14.33 μmol·L^-1, curcumin 4 352.02 μmol·L^-1, tanshinone IIA 1 772.97 μmol·L^-1, baicalein 16.01 μmol·L^-1, and soliquiritigenin 78.48 μmol·L^-1, and CRA recurrence organoids had higher sensitivity to emodin and baicalein. Conclusion The CRA recurrence organoids were successfully constructed to maintain the histological and genetic characteristics of the source tissues. Evaluated by the activities of CRA recurrence organoids, emodin and baicalein could be used as the effective ingredients for the treatment of CRA recurrence. These results provided theoretical and methodological references for the effectiveness of traditional Chinese medicine (TCM) in the treatment of CRA recurrence and the prevention of colorectal cancer (CRC).

R965

国家自然科学基金青年基金项目（82305156，82205024）；江苏省自然科学基金青年基金项目（BK20230162）；江苏省省级重点研发计划（社会发展）项目（BE2021611）；南京市博士后优秀科研项目（NJSBSH2023）