Malic enzyme 1 is important for uterine decidualization in response to progesterone/cAMP/PKA/HB‐EGF pathway

Abstract

Malic enzyme 1 (Me1), a member of the malic enzymes involving in glycolytic pathway and citric acid cycle, is essential for the energy metabolism and maintenance of intracellular redox balance state, but its physiological role and regulatory mechanism in the uterine decidualization are still unknown. Current study showed that Me1 was strongly expressed in decidual cells, and could promote the proliferation and differentiation of stromal cells followed by an accelerated cell cycle transition, indicating an importance of Me1 in the uterine decidualization. Silencing of Me1 attenuated NADPH generation and reduced GR activity, while addition of NADPH improved the defect of GR activity elicited by Me1 depletion. Further analysis found that Me1 modulated intracellular GSH content via GR. Meanwhile, Me1 played a role in maintaining mitochondrial function as indicated by these observations that blockadge of Me1 led to the accumulation of mitochondrial O2- level and decreased ATP production and mtDNA copy numbers accompanied with defective mitochondrial membrane potential. In uterine stromal cells, progesterone induced Me1 expression through PR‐cAMP‐PKA pathway. Knockdown of HB‐EGF might impede the regulation of progesterone and cAMP on Me1. Collectively, Me1 is essential for uterine decidualization in response to progesterone/cAMP/PKA/HB‐EGF pathway and plays an important role in preventing mitochondrial dysfunction.

TAZ as a novel regulator of oxidative damage in decidualization via Nrf2/ARE/Foxo1 pathway

“…Mouse uteri from days 1–8 of pregnancy and undergoing artificial decidualization were gathered as described previously 17 . All animal experimental procedures were approved by the Committee for the Ethics on Animal Care and Use of Jilin University (SY201905031).…”

“…In situ hybridization was performed as depicted previously 17 . Briefly, a TAZ cRNA probe was labeled with digoxigenin and then used for hybridization in frozen sections.…”

“…In vitro decidualization was performed by replenishing estradiol-17β (10 nM, Sigma) plus progesterone (1 mΜ, Sigma) in uterine stromal cells obtained on the 4th day of pregnancy as depicted previously 17 .…”

“…Decidual cells obtained on day 7 of pregnancy were seeded on glass coverslips to perform immunofluorescence as depicted previously 17 . Briefly, after fixation with 4% cold paraformaldehyde, the cells were incubated overnight with antibody against TAZ (1:100, Thermo Fisher Scientific) followed by the addition of goat anti-rabbit Alex Fluor 488 conjugated antibody (1:1000, Invitrogen) and DAPI counterstaining of the nuclei.…”

“…Plasmids overexpressing TAZ variant TAZa and TAZb were constructed as described previously 17 , and the primer sequences were provided in Supplementary Table 1 . Transfection of this overexpression plasmid was performed with Lipofectamine 3000 according to the manufacturer’s protocol.…”