南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (9): 2046-2055.doi: 10.12122/j.issn.1673-4254.2026.09.05

• • 上一篇    

CaMKKβ/AMPK/HIF-1α信号通路通过调控线粒体自噬诱导子宫内膜异位症血管生成

陈元欢1(), 岳斌1, 武权生1(), 毛海燕2, 吉秀家3, 褚翠君1, 陈琳1, 张雨知1, 李冰霞1, 张小花1, 冯鹏4   

  1. 1.甘肃中医药大学，中医临床学院
    2.甘肃省人民医院中医二科，甘肃 兰州 730030
    3.甘肃中医药大学，护理学院，甘肃 兰州 730030
    4.河西学院医学院，甘肃 张掖 734000
  • 收稿日期:2025-12-16 出版日期:2026-09-20 发布日期:2026-09-30
  • 通讯作者: 武权生 E-mail:2334347343@qq.com;wqslanzhou@126.com
  • 作者简介:陈元欢，在读博士研究生，E-mail: 2334347343@qq.com
  • 基金资助:
    国家自然科学基金(82260949);国家自然科学基金(82560954);“岐黄英才”导师专项基金(ZYXKBD-202405);甘肃中医药大学科学研究与创新基金项目(2023KCZD-10);甘肃省研究生“创新之星”项目(2025CXZX-912);甘肃省中医药科研课题(GZKG-2020-49);兰州市科技发展指导计划项目(2023-ZD-237)

Activation of the CaMKKβ/AMPK/HIF-1α signaling pathway promotes angiogenesis in endometriosis by regulating mitophagy

Yuanhuan CHEN1(), Bin YUE1, Quansheng WU1(), Haiyan MAO2, Xiujia JI3, Cuijun CHU1, Lin CHEN1, Yuzhi ZHANG1, Bingxia LI1, Xiaohua ZHANG1, Peng FENG4   

  1. 1.Clinical School of Chinese Medicine
    2.Second Department of Traditional Chinese Medicine, Gansu Provincial People's Hospital, Lanzhou 730030, China
    3.College of Nursing, Gansu University of Traditional Chinese Medicine, Lanzhou 730030, China
    4.Medical College, Hexi University, Zhangye 734000, China
  • Received:2025-12-16 Online:2026-09-20 Published:2026-09-30
  • Contact: Quansheng WU E-mail:2334347343@qq.com;wqslanzhou@126.com
  • Supported by:
    National Natural Science Foundation of China(82260949)

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摘要：

目的 探讨CaMKKβ/AMPK/HIF-1α信号通路是否通过调控线粒体自噬参与子宫内膜异位症（EMs）血管生成的分子机制。 方法 分别培养EMs患者的在位内膜间质细胞（hEM15A）与非EMs女性的正常内膜间质细胞（HESCs）。通过CCK-8法、划痕实验及体外血管生成实验分别检测细胞的增殖、侵袭/迁移及成管能力。采用Western blotting和qPCR技术检测CaMKKβ、AMPK、HIF-1α、BNIP3、LC3、VEGF及CD31的蛋白与mRNA表达水平。使用MitoTracker Green、MitoSOX Red荧光探针及免疫荧光染色观察线粒体形态、mtROS水平及BNIP3/LC3共定位情况。通过使用CaMKKβ激动剂和抑制剂进行干预，以验证该通路的功能。 结果 与HESCs相比，hEM15A细胞表现出更强的增殖、侵袭、迁移及血管生成能力（P<0.01），且CaMKKβ、AMPK、HIF-1α、BNIP3、LC3及VEGF、CD31表达上调（P<0.01）。线粒体形态观察显示EMs细胞线粒体数量增多、结构完整，但BNIP3与LC3的定位信号增强，同时mtROS水平升高（P<0.01）。机制上，抑制CaMKKβ可降低AMPK、BNIP3、LC3、HIF-1α、VEGF、CD31的表达（P<0.01）；反之，激动CaMKKβ则能逆转上述效应，促进线粒体自噬并诱导血管生成（P<0.01）。 结论 在EMs的发生过程中，CaMKKβ/AMPK的活化被异常激活，导致HIF-1α过度积累，进而升高其下游靶基因BNIP3的表达，继而促进线粒体自噬的发生，从而诱导异常血管生成，抑制CaMKKβ的表达则可抑制EMs的疾病进程。

关键词: 子宫内膜异位症, 血管生成, 线粒体自噬, CaMKKβ/AMPK/HIF-1α信号通路

Abstract:

Objective To investigate whether the CaMKKβ/AMPK/HIF-1α signaling pathway participates in angiogenesis in endometriosis by regulating mitophagy. Methods Human endometrial stromal cells (hEM15A) isolated from endometriosis patients and normal endometrial stromal cells (HESCs) from non-endometriosis women were tested for proliferation, invasion, migration and tube formation abilities using CCK-8 assay, scratch assay and in vitro angiogenesis test. The protein and mRNA expression levels of CaMKKβ, AMPK, HIF-1α, BNIP3, LC3, VEGF and CD31 in the cells were detected by Western blotting and qPCR. MitoTracker Green, MitoSOX Red fluorescent probes and immunofluorescence staining were used to observe mitochondrial morphology, mtROS level and BNIP3/LC3 co-localization. CaMKKβ agonists and inhibitors were used to verify the function of the CaMKKβ/AMPK/HIF-1α pathway. Results Compared with HESCs, hEM15A cells showed stronger proliferation, invasion, migration and angiogenesis abilities with significantly up-regulated expressions of CaMKKβ, AMPK, HIF-1α, BNIP3, LC3, VEGF and CD31. hEM15A cells showed increased number of mitochondria with intact structure, but the localization signals of BNIP3 and LC3 and mtROS level increased significantly. Mechanistically, inhibition of CaMKKβ significantly reduced expressions of AMPK, BNIP3, LC3, HIF-1α, VEGF and CD31, while activation of CaMKKβ increased expressions these proteins, down-regulated mitochondrial autophagy and inhibited angiogenesis. Conclusion CaMKKβ/AMPK is abnormally activated in endometriosis to result in excessive accumulation of HIF-1α, which in turn increases the expression of its downstream target gene BNIP3, thus promoting mitophagy to trigger abnormal angiogenesis, which can be inhibited by inhibiting abnormal activation of CaMKKβ.

Key words: endometriosis, angiogenesis, mitochondrial autophagy, CaMKKβ/AMPK/HIF-1α signaling pathway