目的 探讨丹参酮II_A通过调控蛋白激酶B（Akt1）/铁氧化还原蛋白1（FDX1）信号抑制卵巢颗粒细胞的铜死亡，改善环磷酰胺诱导的大鼠卵巢早衰的作用机制。方法 将49只雌性成年SD大鼠随机分为对照组、模型组、丹参酮II_A（10、20 mg/kg）组、丹参酮II_A＋A-674563组、丹参酮II_A＋siNC组、丹参酮II_A＋siFDX1组。采用酶联免疫吸附测定（ELISA）检测血清雌二醇（E₂）和卵泡刺激素（FSH）的水平以及卵巢组织中二氢硫辛酰胺S-乙酰转移酶（DLAT）、二氢硫辛酰胺S-琥珀酰转移酶（DLST）的水平；原子吸收光谱法测定卵巢铜离子含量；荧光探针法检测脂质活性氧（ROS）的水平；Western blotting法分析p-Akt1/Akt1、p-FDX1/FDX1蛋白的表达；免疫荧光化学法与末端脱氧核苷酸转移酶dUTP缺口末端标记法评估细胞增殖（Ki67阳性率）与凋亡；苏木精-伊红（HE）染色评估卵巢组织病理学变化。结果 与模型组相比，丹参酮II_A各剂量组血清E₂水平和原始卵泡数显著升高、FSH显著降低；铜离子含量、脂质ROS、卵巢细胞凋亡率、组织纤维化/炎症评分均显著降低；p-Akt1/Akt1、p-FDX1/FDX1及Ki67阳性率显著升高（P＜0.05）。与丹参酮II_A 20 mg/kg组相比，丹参酮II_A＋A-674563可剂量相关性逆转上述指标（P＜0.05）。与丹参酮II_A＋siNC组相比，丹参酮II_A＋siFDX1组E₂水平、卵泡数、p-Akt1/Akt1、p-FDX1/FDX1及Ki67阳性率显著降低，而FSH、铜离子含量、脂质ROS、卵巢凋亡率及组织损伤评分均显著升高（P＜0.05）。结论 丹参酮II_A通过激活Akt1/FDX1信号，抑制卵巢颗粒细胞铜死亡及凋亡，改善激素失衡与组织病理损伤，从而缓解卵巢早衰进展。

Objective To investigate the mechanisms of tanshinone II_A regulates Akt1/FDX1 signaling to inhibit cuproptosis in ovarian granulosa cells, thereby ameliorating cyclophosphamide-induced premature ovarian failure in rats. Methods Forty-nine adult female SD rats were randomly divided into control group, model group, tanshinone II_A (10, 20 mg/kg) group, tanshinone II_A+A-674563 group, tanshinone II_A+siNC group, tanshinone II_A+siFDX1 group. The levels of serum E₂, FSH, and the levels of DLAT and DLST in ovarian tissue were detected using ELISA. Ovarian copper ion content was determined by atomic absorption spectrophotometry, ROS levels were assessed using a fluorescence probe, expression of p-Akt1/Akt1 and p-FDX1/FDX1 proteins was analyzed by Western blotting, cell proliferation (Ki67 positivity rate) and apoptosis were evaluated using immunofluorescence chemistry and terminal deoxynucleotidyl transferase dUTP nick end labeling assays, respectively. And ovarian histopathological changes were assessed by HE staining. Results Compared to the model group, the tanshinone II_A group significantly increase serum E₂ levels and primordial follicle count, decreased FSH, copper ion content, lipid ROS, apoptosis rate, and tissue fibrosis/inflammation scores (P < 0.05), while p-Akt1/Akt1 and p-FDX1/FDX1 ratios, and Ki67 positivity rate were significantly increased (P< 0.05). Compared with tanshinone II_A 20 mg/kg group, tanshinone II_A+A-674563 reversed above indicators (P < 0.05). Compared to the tanshinone II_A+siNC group, tanshinone II_A+siFDX1 group were significantly decreased E2 levels, follicle counts, p-Akt1/Akt1, p-FDX1/FDX1 ratios, and Ki67 positivity rates, while FSH, copper ion content, lipid ROS, apoptosis rates, and tissue damage scores were significantly increased (P < 0.05). Conclusion Tanshinone II_A ameliorates premature ovarian progression by activating the Akt1/FDX1 signaling, inhibiting granulosa cell cuproptosis and apoptosis, and improving hormonal imbalance and histopathological damage.

R285.5;R287.4

四川省基层卫生事业发展研究中心科研项目（SWFZ24-Y-32）