Fine particulate matter reduces the pluripotency and proliferation of human embryonic stem cells through ROS induced AKT and ERK signaling pathway

Reprod Toxicol. 2020 Sep:96:231-240. doi: 10.1016/j.reprotox.2020.07.010. Epub 2020 Jul 31.

Abstract

Epidemiological investigations have found that air fine particulate matter (PM) exposure not only causes respiratory and cardiovascular diseases in adults and children, but also affects embryonic development during pregnancy, leading to poor pregnancy outcomes. However, its exact molecular mechanism is still unclear. In this study, human embryonic stem cells (hESCs) were treated with PM at different concentrations then the morphology and proliferation capacity were measured. The mRNA and protein expression of NANOG and OCT4 were detected using quantitative PCR, immunofluorescence, western blotting, and flow cytometry. Reactive oxygen species (ROS) generation and AKT/ERK activation were also measured. Meanwhile, changes in ROS, the expression of NANOG, OCT4, and the AKT/ERK pathways were measured in the hESCs with or without pretreatment of ROS scavenger N-acetylcysteine (NAC) prior to PM exposure. After PM exposure, the proliferation capacity and expression of OCT4 and NANOG at the mRNA and protein levels were downregulated. The ROS level in the hESCs increased after PM exposure, but this increase in ROS was attenuated by pretreatment with NAC. Further analysis showed that the levels of phosphorylated AKT and ERK increased after PM exposure. After pretreatment with NAC, the phosphorylation levels of AKT and ERK, which are crucial for regulating the proliferation, pluripotency, and differentiation of hESC, were significantly attenuated compared with the non-NAC pretreated exposure group. These results suggest that PM exposure may reduce the proliferation and pluripotency of hESC through ROS-mediated AKT/ERK pathways, thereby affecting the long-term development of embryos.

Keywords: AKT/ERK; PM; Pluripotency; ROS; hESCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Air Pollutants / toxicity*
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / drug effects*
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • MAP Kinase Signaling System / drug effects
  • Nanog Homeobox Protein / genetics
  • Nanog Homeobox Protein / metabolism
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Particulate Matter / toxicity*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Reactive Oxygen Species / metabolism

Substances

  • Air Pollutants
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Particulate Matter
  • Reactive Oxygen Species
  • Proto-Oncogene Proteins c-akt