目的 基于调控固醇调节元件结合蛋白 1（SREBP1）/溶质载体家族 7 成员 11（SLC7A11）/谷胱甘肽过氧化 物酶 4（GPX4）轴促进铁死亡，探讨健脾化瘀方含药血清抑制胰腺癌 PANC-1 细胞增殖、侵袭的机制。 方法 将 20 只 SD 大鼠随机分为给药组、对照组（每组 10 只），分别灌胃健脾化瘀方药液（26.8 g·kg-1 ·d-1 ）及生 理盐水（每日 1 次，连续 7 d），制备健脾化瘀方含药血清及空白血清。体外培养 PANC-1 细胞，采用 CCK-8 法检测不同浓度（0%、0.5%、2.5%、5%、10%、15%、20%）健脾化瘀方含药血清干预 24 h 后的细胞存活率， 并选择合适的药物浓度进行后续实验。采用克隆形成实验检测细胞增殖能力；划痕实验检测细胞迁移能力； Transwell 实验检测细胞侵袭能力；荧光探针法检测细胞内中性脂质、活性氧（ROS）及脂质过氧化水平；微量法 检测细胞亚铁离子（Fe2+ ）、谷胱甘肽（GSH）、丙二醛（MDA）水平；RT-qPCR 及 Western Blot 法检测细胞 SREBP1、SLC7A11、GPX4、N-cadherin、E-cadherin、Vimentin mRNA 及蛋白表达水平。结果 （1）与空白对 照组比较，2.5%、5%、10%、15%、20% 健脾化瘀方含药血清可显著抑制 PANC-1 细胞存活率（P＜0.05，P＜ 0.01），且呈浓度依赖性；健脾化瘀方含药血清的 IC50值为 15.40%，选取 5%、10%、15% 含药血清浓度作为健 脾化瘀方低、中、高剂量进行后续实验。（2）与空白对照组比较，健脾化瘀方低、中、高剂量组及吉西他滨 （5 μmol·L-1 ）组 PANC-1 细胞的相对克隆形成数、24 h 细胞迁移率及侵袭细胞数均显著降低（P＜0.05，P＜ 0.01）。与空白对照组比较，健脾化瘀方低、中、高剂量组 PANC-1 细胞中 ROS、Fe2+ 、MDA 水平显著升高 （P＜0.05，P＜0.01），GSH 水平显著降低（P＜0.01），细胞内脂质过氧化水平显著升高（P＜0.05，P＜0.01），中 性脂质积累显著减少（P＜0.05，P＜0.01），SREBP1、GPX4、SLC7A11、N-cadherin、Vimentin mRNA 及蛋白表 达水平显著降低（P＜0.05，P＜0.01），E-cadherin mRNA 及蛋白表达水平显著升高（P＜0.05，P＜0.01）。 结论 健脾化瘀方含药血清可以有效地抑制胰腺癌 PANC-1 细胞的增殖、迁移及侵袭，其作用机制可能与抑 制 SREBP1 表达，减少细胞内脂质累积，诱导胰腺癌细胞铁死亡有关。

Objective To investigate the mechanism by which the medicated serum of Jianpi Huayu Formula （JPHYF） inhibits the proliferation and invasion of pancreatic cancer PANC-1 cells，based on its role in regulating the SREBP1/ SLC7A11/GPX4 axis to promote ferroptosis. Methods Twenty SD rats were randomly divided into a control group and a drug-administered group （n=10 per group）. These rats were intragastrically administered either JPHYF decoction （26.8 g·kg-1 ·d-1 ） or an equal volume of normal saline once daily for 7 consecutive days to prepare JPHYF medicated serum and blank control serum， respectively. PANC-1 cells were cultured in vitro. The CCK-8 assay was used to measure changes in cell viability after 24-hour intervention with different concentrations （0%， 0.5%， 2.5%， 5%， 10%，15%，20%） of JPHYF medicated serum，and appropriate concentrations were selected for subsequent experiments. Colony formation assay was employed to detect cell proliferation capacity；scratch wound healing assay was applied to assess cell migration ability；Transwell assay was applied to evaluate cell invasion ability；fluorescent probes were used to measure intracellular neutral lipid content，reactive oxygen species （ROS） levels，and lipid peroxidation levels； micro-methods were applied to detect cellular ferrous iron （Fe2+ ），glutathione （GSH），and malondialdehyde （MDA） levels；RT-qPCR and Western Blot were performed to determine the mRNA and protein expression levels of SREBP1， SLC7A11，GPX4，N-cadherin，E-cadherin，and Vimentin. Results （1） Compared with the blank control group， JPHYF medicated serum at concentrations of 2.5%，5%，10%，15%，and 20% significantly inhibited the survival rate of PANC-1 cells （P＜0.05，P＜0.01） in a concentration-dependent manner. The IC50 value of JPHYF medicated serum was 15.40%. Consequently，concentrations of 5%，10%，and 15% medicated serum were selected as the low-， medium-，and high-dose JPHYF groups for subsequent experiments.（2） Compared with the blank control group，the relative colony formation number，24-hour cell migration rate，and number of invasive cells were significantly reduced in the JPHYF low-，medium-，and high-dose groups and the gemcitabine group （5 μmol·L-1 ）（P＜0.05，P＜0.01）. Compared with the blank control group， the JPHYF low- ， medium- ， and high-dose groups showed significantly increased ROS，Fe2+，and MDA levels of PANC-1 cells （P＜0.05，P＜0.01），significantly decreased GSH levels （P＜0.01）， significantly elevated intracellular lipid peroxidation levels （P＜0.05， P＜0.01）， and significantly reduced neutral lipid accumulation （P＜0.05， P＜0.01）. Furthermore， the mRNA and protein expression levels of SREBP1，GPX4，SLC7A11，N-cadherin，and Vimentin were significantly decreased （P＜0.05，P＜0.01），while the mRNA and protein expression levels of E-cadherin were significantly increased （P＜0.05，P＜0.01） in the JPHYF treatment groups. Conclusions JPHYF medicated serum can effectively inhibit the proliferation， migration， and invasion of pancreatic cancer PANC-1 cells. Its mechanism of action may be related to the suppression of SREBP1 expression，reduction of intracellular lipid accumulation，and induction of ferroptosis in pancreatic cancer cells.

R285.5

国家自然科学基金项目（82274526）；广东省自然科学基金项目（2023A1515011069）；广州中医药大学“双一流”与高水平大学学科 后备人才培育项目（A1-2601-22-415-023）；中医证候重点实验室科研项目；广州中医药大学第一临床医学院“揭榜挂帅”研究生创新能力提升 项目（A3-0317-25-110-005）。