1. Academic Validation
  2. NETs promote ROS production to induce human amniotic epithelial cell apoptosis via ERK1/2 signaling in spontaneous preterm birth

NETs promote ROS production to induce human amniotic epithelial cell apoptosis via ERK1/2 signaling in spontaneous preterm birth

  • Am J Reprod Immunol. 2022 Nov 21;e13656. doi: 10.1111/aji.13656.
Min Hu 1 2 Hao Li 1 2 Guangzhen Li 3 Yuan Wang 1 2 Jing Liu 1 2 Meihua Zhang 2 Di Shen 4 1 2 Xietong Wang 4 1 2
Affiliations

Affiliations

  • 1 Department of Obstetrics and Gynaecology, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, China.
  • 2 Key Laboratory of Birth Regulation and Control Technology of National Health Commission of China, Shandong Provincial Maternal and Child Health Care Hospital, Jinan, Shandong, China.
  • 3 Department of General Surgery, Qilu Hospital of Shandong University, Jinan, China.
  • 4 Department of Obstetrics and Gynaecology, Maternal and Child Health Care Hospital of Shandong Provincial, Jinan, Shandong, China.
Abstract

Problem: Premature birth is a common obstetric complication but its pathogenesis is unclear. Inflammation at the maternal-fetal interface in preterm labor leads to the infiltration of neutrophils, which promotes inflammatory responses and induces the degradation of extracellular matrix and cell Apoptosis, thus contributing to preterm labor. It is unclear whether neutrophil extracellular traps (NETs), a functional form of neutrophils, are involved in preterm labor.

Methods of study: After collecting amniotic membranes from research objects, we localized NETs by immunofluorescence and evaluated the expression of matrix metalloproteinase (MMP)-9 and MMP-2 by western blotting. Primary human amniotic epithelial cells (hAECs) subjected to treatment with NETs, 5-ethynyl-20-deoxyuridine cell proliferation assay, Lactate Dehydrogenase (LDH) assay, western blotting, and flow cytometry Apoptosis assay were used to determine the effects of NETs on hAECs. We also elucidated possible mechanisms underlying the effects.

Results: Compared with normal term women, NETs infiltration and MMP-9 expression in the amniotic membrane from preterm women had increased. Thereafter, NETs might suppress the proliferation and promote the Apoptosis of hAECs. Furthermore, after NETs treatment, the mitochondrial membrane potential was significantly decreased, ERK1/2 phosphorylation expression was upregulated and Reactive Oxygen Species (ROS) production was increased in hAECs. Changes in cell proliferation, LDH release, and cell Apoptosis level due to NETs could be reversed by ROS inhibitor or ERK phosphorylation inhibitors.

Conclusions: NETs can promote the Apoptosis of hAECs via ERK1/2 pathways dependent on ROS release.

Keywords

ERK1/2 signaling; ROS; apoptosis; human amniotic epithelial cells (hAECs); neutrophil extracellular traps (NETs); spontaneous preterm birth (sPTB).

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