目的 基于“种子-土壤”学说探讨化瘀消癥颗粒调控铁死亡治疗输卵管妊娠（TP）的作用机制。方法 采用 铁死亡诱导剂 Erastin 诱导滋养细胞系 HTR-8/SVneo 细胞及原代输卵管上皮细胞，模拟 TP 铁死亡微环境，给 予化瘀消癥颗粒含药血清干预。采用免疫荧光法检测原代输卵管上皮细胞 PAX8 表达；CCK-8 法检测细胞增 殖活力；Western Blot 法检测细胞中铁死亡相关蛋白（TFRC、ACSL4、GPx4）的表达水平；DCFH-DA 探针检测 细胞内活性氧（ROS）水平。结果 （1）与对照组比较，2 μmol·L-1 Erastin 干预 24 h 后，滋养细胞（HTR-8/ SVneo）的铁死亡相关蛋白 TFRC、ACSL4 表达水平显著升高（P＜0.05，P＜0.01），GPx4 蛋白表达水平显著降低 （P＜0.01），细胞内 ROS 水平显著升高（P＜0.01）。与 Erastin 组比较，Erastin+15% 化瘀消癥颗粒含药血清组 滋养细胞的 TFRC 蛋白表达水平显著升高（P＜0.01），GPx4 蛋白表达水平明显降低（P＜0.05），细胞内 ROS 水 平进一步显著升高（P＜0.001）。（2）与对照组比较，3 μmol·L-1 Erastin 干预 24 h 后，原代输卵管上皮细胞的铁 死亡相关蛋白 TFRC、ACSL4 表达水平显著升高（P＜0.01，P＜0.001），GPx4 蛋白表达水平显著降低（P＜ 0.001），细胞内 ROS 水平显著升高（P＜0.01）。与 Erastin 组比较，Erastin+5% 化瘀消癥颗粒含药血清组输卵管 上皮细胞的 TFRC、ACSL4 蛋白表达水平明显降低（P＜0.05），GPx4 蛋白表达水平显著升高（P＜0.01），细胞内 ROS 水平显著降低（P＜0.01）。结论 化瘀消癥颗粒含药血清一方面能协同 Erastin 促进滋养细胞铁死亡，另一 方面能减轻 Erastin 诱导的输卵管上皮细胞损伤。表明化瘀消癥颗粒可能通过调节“种子（滋养细胞）”和“土 壤（输卵管上皮细胞）”的铁死亡敏感性，发挥治疗 TP 的作用。

Objective To explore the mechanism of Huayu Xiaozheng Granules（HYXZ）on regulating ferroptosis in treating tubal pregnancy（TP）based on the“seed-soil theory”. Methods The ferroptosis inducer erastin was used to induce trophoblast cell line HTR-8/SVneo and primary fallopian tube epithelial cells and to simulate TP ferroptosis microenvironment. HYXZ-containing serum was used to intervene in cells. PAX8 expression in primary fallopian tube epithelial cells was detected by immunofluorescence assay. Cell proliferation activity was analyzed by CCK-8， the expressions of ferroptosis related proteins（TFRC，ACSL4 and GPx4）were tested by Western Blot，and intracellular reactive oxygen species（ROS）were detected by DCFH-DA probe. Results （1）Compared with control group，after 24-h intervention of 2 μmol·L-1 Erastin，the expressions of ferroptosis related proteins TFRC and ACSL4 in HTR-8/SVneo increased significantly（P＜0.05，P＜0.01），while GPx4 expression level obviously reduced（P＜0.01），intracellular ROS level increased significantly（P＜0.001）. Compared with Erastin group，the protein expression level of TFRC in HTR-8/SVneo remarkably increased（P＜0.01）， but the protein expression level of GPx4 obviously reduced（P＜ 0.01）， intracellular ROS level significantly increased（P＜0.001）in Erastin+15% HYXZ-containing serum group. （2）Compared with control group，after 24-h intervention of 3 μmol·L-1 Erastin，the expressions of ferroptosis related proteins TFRC and ACSL4 in primary fallopian tube epithelial cells increased significantly（P＜0.01， P＜0.001）， while GPx4 expression level obviously reduced（P＜0.001），intracellular ROS level increased significantly（P＜0.01）. Compared with Erastin group， the protein expression level of TFRC and ACSL4 in fallopian tube epithelial cells decreased obviously（P＜0.05），while GPx4 expression level obviously increased（P＜0.01），intracellular ROS level decreased significantly（P＜0.01）in Erastin+5% HYXZ-containing serum group. Conclusion HYXZ-containing serum can cooperate with Erastin to promote trophoblast ferroptosis and protect fallopian tube epithelial cells from Erastin damage. The results showed that HYXZ achieve the therapeutic effect of TP by regulating the ferroptosis sensitivity of “seed（trophoblast）” and “soil（fallopian tube epithelial cells）”.

R285.5

国家自然科学基金项目（82174417）