目的 構(gòu)建人源結(jié)直腸腺瘤（CRA）復(fù)發(fā)類器官，評價幾種中藥活性成分對CRA復(fù)發(fā)類器官活性的影響。方法 收集內(nèi)鏡下切除后再發(fā)腺瘤的CRA患者組織，腺瘤數(shù)量數(shù)目超過2枚且少于6枚，直徑為5～10 mm，構(gòu)建CRA復(fù)發(fā)三維（3D）類器官；使用高內(nèi)涵智能分析儀和光學(xué)顯微鏡觀察類器官形態(tài)；運(yùn)用蘇木精-伊紅（HE）染色觀察類器官組織學(xué)形態(tài)；采用外顯子組測序檢測CRA復(fù)發(fā)類器官與源組織基因?qū)W特征的一致性；采用CellTiter-Glo檢測1 000.00、100.00、10.00、1.00、0.10、0.01 μmol·L^-1的鹽酸小檗堿、人參皂苷Rg₃、大黃素、姜黃素、丹參酮Ⅱ A、黃芩素、異甘草素對6例CRA復(fù)發(fā)類器官活性的影響，繪制活性曲線，并計算半數(shù)抑制濃度（IC₅₀）值。結(jié)果 建成CRA復(fù)發(fā)類器官，形態(tài)多以內(nèi)部透亮、近圓的囊樣結(jié)構(gòu)為主，且囊壁多有向外側(cè)隆起的蕈狀結(jié)構(gòu)；經(jīng)過多次傳代、凍存再復(fù)蘇后其形態(tài)結(jié)構(gòu)均能保持一致；每組類器官與源組織之間的突變類型比例基本一致，插入和缺失突變的長度和比例、拷貝變異數(shù)均具有較高度相似性； 7種中藥活性成分對6例CRA復(fù)發(fā)類器官的IC₅₀均值分別為鹽酸小檗堿69.79 μmol·L^-1、人參皂苷Rg₃ 110.92 μmol·L^-1、大黃素14.33 μmol·L^-1、姜黃素4 352.02 μmol·L^-1、丹參酮Ⅱ A 1 772.97 μmol·L^-1、黃芩素16.01 μmol·L^-1、異甘草素78.48 μmol·L^-1，CRA復(fù)發(fā)類器官對大黃素和黃芩素具有更高的敏感性。結(jié)論 成功構(gòu)建能夠穩(wěn)定傳代、凍存和復(fù)蘇的CRA復(fù)發(fā)類器官，其能維持源組織的組織學(xué)和基因?qū)W特征；經(jīng)CRA復(fù)發(fā)類器官活性評價，大黃素和黃芩素具有治療CRA復(fù)發(fā)的潛力，為中醫(yī)藥在治療CRA復(fù)發(fā)和防治結(jié)直腸癌（CRC）領(lǐng)域的有效應(yīng)用提供理論和方法學(xué)參考。

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

國家自然科學(xué)基金青年基金項目（82305156，82205024）；江蘇省自然科學(xué)基金青年基金項目（BK20230162）；江蘇省省級重點研發(fā)計劃（社會發(fā)展）項目（BE2021611）；南京市博士后優(yōu)秀科研項目（NJSBSH2023）