Type 2 diabetes mellitus induces congenital heart defects in murine embryos by increasing oxidative stress, endoplasmic reticulum stress, and apoptosis

Am J Obstet Gynecol. 2016 Sep;215(3):366.e1-366.e10. doi: 10.1016/j.ajog.2016.03.036. Epub 2016 Mar 31.

Abstract

Background: Maternal type 1 and 2 diabetes mellitus are strongly associated with high rates of severe structural birth defects, including congenital heart defects. Studies in type 1 diabetic embryopathy animal models have demonstrated that cellular stress-induced apoptosis mediates the teratogenicity of maternal diabetes leading to congenital heart defect formation. However, the mechanisms underlying maternal type 2 diabetes mellitus-induced congenital heart defects remain largely unknown.

Objective: We aim to determine whether oxidative stress, endoplasmic reticulum stress, and excessive apoptosis are the intracellular molecular mechanisms underlying maternal type 2 diabetes mellitus-induced congenital heart defects.

Study design: A mouse model of maternal type 2 diabetes mellitus was established by feeding female mice a high-fat diet (60% fat). After 15 weeks on the high-fat diet, the mice showed characteristics of maternal type 2 diabetes mellitus. Control dams were either fed a normal diet (10% fat) or the high-fat diet during pregnancy only. Female mice from the high-fat diet group and the 2 control groups were mated with male mice that were fed a normal diet. At E12.5, embryonic hearts were harvested to determine the levels of lipid peroxides and superoxide, endoplasmic reticulum stress markers, cleaved caspase 3 and 8, and apoptosis. E17.5 embryonic hearts were harvested for the detection of congenital heart defect formation using India ink vessel patterning and histological examination.

Results: Maternal type 2 diabetes mellitus significantly induced ventricular septal defects and persistent truncus arteriosus in the developing heart, along with increasing oxidative stress markers, including superoxide and lipid peroxidation; endoplasmic reticulum stress markers, including protein levels of phosphorylated-protein kinase RNA-like endoplasmic reticulum kinase, phosphorylated-IRE1α, phosphorylated-eIF2α, C/EBP homologous protein, and binding immunoglobulin protein; endoplasmic reticulum chaperone gene expression; and XBP1 messenger RNA splicing, as well as increased cleaved caspase 3 and 8 in embryonic hearts. Furthermore, maternal type 2 diabetes mellitus triggered excessive apoptosis in ventricular myocardium, endocardial cushion, and outflow tract of the embryonic heart.

Conclusion: Similar to those observations in type 1 diabetic embryopathy, maternal type 2 diabetes mellitus causes heart defects in the developing embryo manifested with oxidative stress, endoplasmic reticulum stress, and excessive apoptosis in heart cells.

Keywords: apoptosis; endoplasmic reticulum stress; heart defects; oxidative stress; type 2 diabetes mellitus.

MeSH terms

  • Animals
  • Apoptosis*
  • Caspase 3 / metabolism
  • Caspase 8 / metabolism
  • Diabetes Mellitus, Experimental
  • Diabetes, Gestational*
  • Embryo, Mammalian
  • Endoplasmic Reticulum / metabolism
  • Endoplasmic Reticulum Chaperone BiP
  • Endoplasmic Reticulum Stress*
  • Endoribonucleases / metabolism
  • Female
  • Heart Defects, Congenital / embryology*
  • Heart Defects, Congenital / pathology
  • Heat-Shock Proteins / metabolism
  • Lipid Peroxidation
  • Mice, Inbred C57BL
  • Myocardium / metabolism
  • Myocardium / pathology
  • Oxidative Stress*
  • Phosphorylation
  • Pregnancy
  • Protein Kinases / metabolism
  • Protein Serine-Threonine Kinases / metabolism
  • RNA Splicing
  • Transcription Factor CHOP / metabolism
  • X-Box Binding Protein 1 / genetics

Substances

  • Endoplasmic Reticulum Chaperone BiP
  • Heat-Shock Proteins
  • X-Box Binding Protein 1
  • Xbp1 protein, mouse
  • Transcription Factor CHOP
  • Protein Kinases
  • Ern1 protein, mouse
  • Protein Serine-Threonine Kinases
  • eIF2alpha kinase, mouse
  • Endoribonucleases
  • Casp8 protein, mouse
  • Caspase 3
  • Caspase 8