目的 探讨波棱芴酮促进诱导多能干细胞（induced pluripotent stem cells，iPSCs）向肝样细胞（hepatocyte-like cells，HLCs）分化及其协同修复药源性肝损伤（drug-induced liver injury，DILI）的作用。方法 通过碱性磷酸酶染色鉴定iPSCs，CCK-8法确定波棱芴酮安全浓度后，使用波棱芴酮和细胞生长因子分别诱导iPSCs分化，通过免疫荧光染色、qRT-PCR等考察波棱芴酮对iPSCs的诱导作用。构建DILI小鼠模型，设置对照组、模型组、iPSCs组和iPSCs＋波棱芴酮（80 mg/kg）组，采用苏木素-伊红染色观察肝组织病理变化，检测血清中肝功能相关生化指标水平，采用免疫荧光染色检测人甲胎蛋白和增殖细胞核抗原的表达。结果 体外实验结果显示，在波棱芴酮的诱导下，iPSCs中肝脏标志物白蛋白、甲胎蛋白、α1-抗胰蛋白酶、细胞角蛋白18、精氨琥珀酸合成酶1、氨甲酰磷酸合成酶1、细胞色素P450 3A4、细胞色素P450 3A7等基因的表达显著升高（P＜0.01），八聚体结合转录因子4的表达降低（P＜0.01），糖原和脂质的储存能力增强（P＜0.01）。体内实验结果显示，波棱芴酮与iPSCs的联合治疗改善了DILI模型小鼠肝脏损伤，显著降低了丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、碱性磷酸酶、总胆红素、直接胆红素、总胆汁酸的水平（P＜0.05），升高白蛋白水平（P＜0.05）；波棱芴酮能够促进DILI小鼠iPSCs细胞的定向迁移和分化，升高人甲胎蛋白的水平，上调增殖细胞核抗原的表达（P＜0.01）。结论 波棱芴酮能够有效诱导iPSCs向HLCs分化，并能够在体内与iPSCs协同发挥改善DILI作用。

Objective To investigate the role of herpetfluorenone in promoting the differentiation of induced pluripotent stem cells (iPSCs) into hepatocyte-like cells (HLCs) and their synergistic effects in repairing drug-induced liver injury (DILI). Methods iPSCs were identified by alkaline phosphatase staining, and the safe concentration of herpetfluorenone was determined by CCK-8 method. Then, herpetfluorenone and cell growth factor were used to induce differentiation of iPSCs. The induction effects of herpetfluorenone on iPSCs was investigated by immunofluorescence staining, qRT-PCR and other methods. A mouse DILI model was constructed, control group, model group, iPSCs group and iPSCs + herpetfluorenone (80 mg/kg) group were set up. The pathological changes of liver tissue were observed using hematoxylin eosin staining, the levels of liver function related biochemical indicators in serum were detected, the expressions of human alpha fetoprotein and proliferating cell nuclear antigen were detected using immunofluorescence staining. Results The results of the in vitro experiments showed that under the induction of herpetfluorenone, the expressions of liver-related genes such as albumin, alpha fetoprotein, alpha-1-antitrypsin, cytokeratin 18, argininosuccinate synthase 1, carbamoyl phosphate synthetase 1, cytochrome P450 3A4, and cytochrome P450 3A7 in iPSCs were significantly increased (P < 0.01), while the expression of octamer-binding transcription factor 4 was decreased (P < 0.01), the storage capacity of glycogen and lipids has been enhanced (P < 0.01). The results of the in vivo experiments showed that the combined treatment of herpetfluorenone and iPSCs improved liver injury in DILI model mice, significantly reduced the levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, direct bilirubin and total bile acid (P < 0.05), and increased the level of albumin (P < 0.05). Herpetfluorenone promoted the directed migration and differentiation of iPSCs in DILI mice, increased the level of human alpha-fetoprotein, and upregulated the expression of proliferating cell nuclear antigen (P < 0.01). Conclusion Herpetfluorenone could effectively induce the differentiation of iPSCs into HLCs and synergistically improve DILI in vivo with iPSCs.

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四川省中医药管理局中医药科研专项（2024ZD026）；国家自然科学基金面上项目（82174083）；西南民族大学青藏高原研究科技创新团队（2024CXTD16）