采用液滴重叠法构建类器官3D培养模型,评价何首乌易感物质顺式二苯乙烯苷(cis-SG)的肝损伤作用。结果表明,相对于普通2D培养肝细胞模型,所构建的类器官3D培养模型的L02细胞和HepG2细胞白蛋白表达分别提高2.5和6.7倍;在第21天时,尿素生成水平分别提高8.3和15.5倍,且HepG2细胞构建的类器官模型显著优于L02细胞;相对于2D培养肝细胞模型,HepG2细胞构建的类器官模型的药物Ⅰ相和Ⅱ相代谢酶表达量显著上调,如CYP2C9、CYP3A4和CYP2D6表达上调分别为381.9、87.0和312.6倍,药物转运体相关基因也明显上调,提示所建立的类器官3D培养模型的肝脏合成和代谢功能显著优于普通2D培养肝细胞模型。肝毒性评价结果表明,相对于普通2D培养肝细胞模型,类器官3D培养模型可以更灵敏地评价对乙酰氨基酚等肝毒性阳性药物的毒性差异,且在类器官3D培养模型上重复给药评价的半数抑制浓度(IC_(50))值显著低于单次给药方式;何首乌易感物质cis-SG在普通2D培养肝细胞模型未检测到IC_(50),在类器官3D培养模型上单次给药方式的IC_(50)是肝毒性阳性药环孢霉素的1.9倍,而其光学异构体反式二苯乙烯苷(trans-SG)的IC_(50)比cis-SG高4.1倍,与前期整体动物水平的毒性强弱基本一致;cis-SG在类器官3D培养模型上多次给药方式的IC_(50)进一步降低,提示长期用药可能增大何首乌肝损伤风险。综上,何首乌易感物质cis-SG对类器官的肝毒性大于trans-SG;类器官3D培养模型可长期培养且保留更多的肝脏合成和代谢功能,适用于中药成分低浓度、长时间暴露产生肝毒性评价和机制研究。 The 3D organotypic organogenesis model was established by drop overlap method to evaluate the hepatic injury induced by cis-SG, a susceptible substance of Radix Polygoni Multiflori. The results showed that the expression of albumin in L02 cells and HepG2 cells increased by 2.5 and 6.7 times, respectively, compared with the normal 2D cultured hepatocytes model. The urea production level increased by 8.3 and 15.5 times respectively on the 21st day , And the organotypic model constructed by HepG2 cells was significantly better than that of L02 cells. Compared with the 2D cultured hepatocyte model, the expression levels of phase I and phase Ⅱ metabolic enzymes in organ-like organs constructed by HepG2 cells were significantly increased, such as CYP2C9, CYP3A4 and CYP2D6 The up-regulation was up to 381.9, 87.0 and 312.6 respectively, and the related genes of drug transporter were also significantly up-regulated, which indicated that the liver synthesis and metabolism function of the established organoid 3D culture model was significantly better than the normal 2D culture hepatocyte model. The results of hepatotoxicity evaluation showed that the organoid 3D culture model can more sensitively evaluate the toxicity difference of the hepatotoxicity-positive drugs such as acetaminophen, compared with the common 2D culture hepatocyte model, and evaluate the repeated administration in the organoid 3D culture model (IC 50) was significantly lower than that of single administration. The cis-SG, a susceptible substance of Polygonum multiflorum, did not detect IC 50 in the normal 2D culture hepatocyte model, IC50 of sub-administration was 1.9 times higher than that of cyclosporine, a positive hepatotoxic drug, and the IC50 of trans-SG was higher than that of cis-SG Fold, which is consistent with that of the whole animal in the previous period. The IC50 of multiple administrations of cis-SG in the organoid 3D culture model is further reduced, suggesting that long-term administration may increase the risk of hepatic injury. In conclusion, cis-SG, a susceptible substance of Polygonum multiflorum, has a higher hepatotoxicity than trans-SG on organ-like organs. 3D organotypic organotypic models can be cultured for long term and retain more liver synthesis and metabolism functions. It is suitable for low concentrations and prolonged exposure of traditional Chinese medicine Assessment of hepatotoxicity and its mechanism.