Scholarlink

Objective <p>This study investigated whether the mechanism by which diethylstilbestrol (DES) causes abnormal development of the gubernaculum testis is related to the AMP-activated protein kinase (AMPK) pathway, from the perspective of DES-induced oxidative stress inhibiting the proliferation and promoting the apoptosis of gubernaculum testis cells in mice.</p> Methods <p>Pregnant mice received subcutaneous DES (0.02, 0.5, 25&#xa0;µg/kg/day) from gestational day 9 to 17 to assess offspring gubernaculum testis development. Immunofluorescence, immunohistochemistry and western blot were used to detect proliferation (BrdU, Ki-67, PCNA), apoptosis (caspase-3, Bax, Bcl-2), AMPK/p-AMPK, and oxidative stress (gp91phox, p47phox) markers in gubernaculum testis tissues. Primary gubernaculum testis cells were cultured and divided into control, DES (low/medium/high), AMPK inhibitor (Compound C), and ROS scavenger (NAC) intervention groups. CCK-8, BrdU, TUNEL, flow cytometry, and DHE staining were used to detect cell proliferation, apoptosis, cycle and intracellular ROS. Western blot analyzed related protein and AMPK/p-AMPK expression.</p> Results <p>DES increased mouse stillbirth rate in a dose-dependent manner (<i>P</i> &lt; 0.05 for medium/high doses), impaired gubernaculum testis development and inhibited testicular descent. DES suppressed proliferation markers but upregulated apoptosis and oxidative stress markers in gubernaculum testis tissues. In vitro, DES elevated intracellular ROS, induced G<sub>1</sub>/S phase arrest and apoptosis, and inhibited proliferation. DES also downregulated p-AMPK; Compound C further exacerbated oxidative stress, while NAC reversed DES-induced oxidative stress and restored AMPK phosphorylation, indicating a bidirectional regulation between AMPK and oxidative stress.</p> Conclusion <p>DES inhibits the AMPK pathway, relieving its negative regulation of NADPH oxidase and causing excessive ROS production and oxidative stress; accumulated oxidative stress feedback inhibits AMPK phosphorylation, forming a vicious cycle that ultimately leads to gubernacular cell injury.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Role of AMP-activated protein kinase in oxidative damage of the gubernaculum testis induced by diethylstilbestrol

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月7日

查看全文
  • Shuiping Tang,
  • Yao Xie,
  • Hanyue Ding,
  • Anqi Huang,
  • Lingjuan Weng,
  • Maxian Fu,
  • Zhuo Li,
  • Lijun Ren,
  • Weifang Wu,
  • Kehua Huang,
  • Shouxing Duan

摘要

Objective

This study investigated whether the mechanism by which diethylstilbestrol (DES) causes abnormal development of the gubernaculum testis is related to the AMP-activated protein kinase (AMPK) pathway, from the perspective of DES-induced oxidative stress inhibiting the proliferation and promoting the apoptosis of gubernaculum testis cells in mice.

Methods

Pregnant mice received subcutaneous DES (0.02, 0.5, 25 µg/kg/day) from gestational day 9 to 17 to assess offspring gubernaculum testis development. Immunofluorescence, immunohistochemistry and western blot were used to detect proliferation (BrdU, Ki-67, PCNA), apoptosis (caspase-3, Bax, Bcl-2), AMPK/p-AMPK, and oxidative stress (gp91phox, p47phox) markers in gubernaculum testis tissues. Primary gubernaculum testis cells were cultured and divided into control, DES (low/medium/high), AMPK inhibitor (Compound C), and ROS scavenger (NAC) intervention groups. CCK-8, BrdU, TUNEL, flow cytometry, and DHE staining were used to detect cell proliferation, apoptosis, cycle and intracellular ROS. Western blot analyzed related protein and AMPK/p-AMPK expression.

Results

DES increased mouse stillbirth rate in a dose-dependent manner (P < 0.05 for medium/high doses), impaired gubernaculum testis development and inhibited testicular descent. DES suppressed proliferation markers but upregulated apoptosis and oxidative stress markers in gubernaculum testis tissues. In vitro, DES elevated intracellular ROS, induced G1/S phase arrest and apoptosis, and inhibited proliferation. DES also downregulated p-AMPK; Compound C further exacerbated oxidative stress, while NAC reversed DES-induced oxidative stress and restored AMPK phosphorylation, indicating a bidirectional regulation between AMPK and oxidative stress.

Conclusion

DES inhibits the AMPK pathway, relieving its negative regulation of NADPH oxidase and causing excessive ROS production and oxidative stress; accumulated oxidative stress feedback inhibits AMPK phosphorylation, forming a vicious cycle that ultimately leads to gubernacular cell injury.