Page 20 - 南京医科大学自然版

南京医科大学学报（自然科学版） 第46卷第10期
·1450 · Journal of Nanjing Medical University（Natural Sciences） 2026年10月

·专题研究：生殖医学·

过氧化物还原酶4蛋白在衰老小鼠卵巢颗粒细胞中的核转位研究

段学诚 ，孟 艳 1，2，3*
1，2
南京医科大学生殖医学与子代健康全国重点实验室，江苏 南京 211166；南京医科大学第一附属医院生殖医学中心，江
1 2
苏 南京 210036；宿迁市第一人民医院（南京医科大学附属宿迁医院），江苏 宿迁 223800
3

［摘 要］ 目的：探讨衰老状态下卵巢颗粒细胞（granulosa cell，GC）中过氧化还原酶4（peroxiredoxin 4，PRDX4）的核转位现象
与细胞衰老表型的关系。方法：分别选取6周龄（年轻组）和9月龄（老龄组）的雌性C57BL/6小鼠，取卵巢组织进行免疫荧光染
色，观察PRDX4在GC中的核转位现象。同时培养人卵巢颗粒细胞系KGN，并采用毒胡萝卜素（thapsigargin，Tg）诱导建立体外
衰老模型。通过CCK⁃8法筛选较优诱导条件，利用Western blot 和RT⁃qPCR 检测衰老相关蛋白（P16、P21、P53）及衰老相关分
泌表型（senescence⁃associated secretory phenotype，SASP）因子［白介素（interleukin，IL）⁃1α、IL⁃1β、IL⁃6、IL⁃8）］的表达水平，以验
证衰老模型。在此基础上，进一步验证衰老表型对PRDX4核转位的影响。结果：免疫荧光结果显示，与年轻组相比，老龄组小
鼠卵巢GC中PRDX4细胞核内的荧光信号显著增强（P < 0.01）。在体外实验中，正常KGN细胞中PRDX4主要定位于胞质；经
Tg 诱导衰老后，衰老组中 P16、P21、P53 蛋白表达水平显著升高（P 均<0.05），SASP 相关因子 mRNA 表达水平也显著上调（P <
0.001），证实衰老模型构建成功。核质分离实验进一步表明，衰老组细胞核中PRDX4蛋白含量较年轻组显著增加（P < 0.01），
而胞质中相对应减少，表明PRDX4由胞质向细胞核转位。结论：衰老可诱导卵巢GC中PRDX4由胞质向细胞核转位，其核转
位现象的改变可能参与调控卵巢衰老进程。
［关键词］ 卵巢衰老；颗粒细胞；细胞衰老；PRDX4；核转位
［中图分类号］ R714.8 ［文献标志码］ A ［文章编号］ 1007⁃4368（2026）10⁃1450⁃07
doi：10.7655/NYDXBNSN260729

Nuclear translocation of peroxiredoxin 4 protein in ovarian granulosa cells of aging mice
1，2
Duan Xuecheng ，Meng Yan 1，2，3*
State Key Laboratory of Reproductive Medicine and Offspring Health，Nanjing Medical University，Nanjing 211166；
1
2 Clinical Center of Reproductive Medicine，the First Affiliated Hospital of Nanjing Medical University，Nanjing
210036；Suqian First Hospital（Suqian Hospital Affiliated to Nanjing Medical University），Suqian 223800，China
3

［Abstract］ Objective： To investigate the relationship between the nuclear translocation of peroxiredoxin 4（PRDX4）in ovarian
granulosa cells（GCs）and the cell senescence phenotype in aging state. Methods： The ovarian tissues of 6⁃week⁃old（young group）and
9⁃month⁃old（aged group）female C57BL/6 mice were taken for immunofluorescence staining to observe the nuclear translocation of
PRDX4 in GCs. At the same time，human ovarian granulosa cell line KGN was cultured，and thapsigargin（Tg）was used to induce the
establishment of in vitro aging model. The optimal induction conditions were screened by CCK ⁃ 8 assay. The expression levels of
senescence⁃related proteins（P16，P21，P53）and senescence⁃associated secretory phenotype（SASP）factors［interleukin（IL）⁃1α，IL⁃
1β，IL⁃6，IL⁃8］were detected by western blot and（real⁃time quantitative PCR，RT⁃qPCR）to validate the aging model. On this basis，
the effect of the senescence phenotype on the nuclear translocation of PRDX4 was further examined. Results： Immunofluorescence
results showed that compared with the young group，the fluorescence signal in the nucleus of PRDX4 in the ovarian GCs of the aged
group was significantly enhanced（P < 0.01）. In vitro，PRDX4 was mainly localized in the cytoplasm of normal KGN cells. After Tg⁃
induced aging，the expression levels of P16，P21，and P53 proteins in the aging group were significantly increased（all P < 0.05），and
the mRNA expression levels of SASP⁃related factors were also significantly up⁃regulated（P < 0.001），confirming that the aging model

［基金项目］ 国家自然科学基金（82271688）
通信作者（Corresponding author），E⁃mail：ctmengyan@hotmail.com（ORCID：0000⁃0001⁃5956⁃8387）
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