title: Up-regulated expression of WNT5a increases inflammation and oxidative stress via PI3K/AKT/NF-κB signaling in the granulosa cells of PCOS patients.
abstract: BACKGROUND
  Polycystic ovary syndrome (PCOS) is a heterogeneous endocrine disorder accompanied by chronic low-grade inflammation, but the molecular mechanism remains unclear.
  
  
  OBJECTIVE
  We investigated the action of WNT5a in the development of chronic inflammation in PCOS and the related molecular signaling pathways.
  
  
  METHODS
  This was a prospective study conducted at the Division of Reproduction Center, Peking University Third Hospital.
  
  
  METHODS
  A total of 35 PCOS patients and 87 control women who reported to the clinic for the in vitro procedure and the cause of marital infertility was male azoospermia were included.
  
  
  METHODS
  Mural granulosa cells (GCs) of 35 PCOS patients and 37 controls were collected during oocyte retrieval and gene expression was analyzed. The human KGN cells and mural GCs from 50 control subjects (six to eight samples were pooled together for each experiment) were cultured in vitro. The regulation of inflammation and oxidative stress was confirmed by quantitative PCR, flow-cytometric assay, and dual-luciferase reporter assay after inflammatory stimuli or WNT5a overexpression. Relevant signaling pathways were identified using specific inhibitors.
  
  
  RESULTS
  Our data demonstrate significantly elevated WNT5a expression in the mural GCs of PCOS patients compared with the controls. Lipopolysaccharide stimulation increased WNT5a expression in KGN cells and mural GCs, and BAY-117082 and pyrrolidinedithiocarbamic acid [nuclear factor-κB (NF-κB) inhibitor] treatments suppressed WNT5a mRNA below the control level. WNT5a overexpression also enhanced the expression of inflammation-related genes and increased intracellular reactive oxygen species, whereas both BAY-117082 and LY-294002 (phosphatidylinositol 3-kinase inhibitor) significantly inhibited WNT5a-induced inflammation and oxidative stress.
  
  
  CONCLUSIONS
  WNT5a acts as a proinflammatory factor in human ovarian GCs. The up-regulated expression of WNT5a in PCOS increases inflammation and oxidative stress predominantly via the phosphatidylinositol 3-kinase/AKT/NF-κB signaling pathway. The proinflammatory cytokines induced might further enhance WNT5a expression via NF-κB-dependent regulation, indicating a novel regulatory system for chronic inflammation in PCOS.
authors.affiliation: Reproductive Medical Center (Y.Z., C.Z., Y.H., Y.Y., R.L., M.L., N.L., P.L., J.Q.), Department of Obstetrics and Gynecology, Peking University Third Hospital, and Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology (Y.Z., Y.H., M.L., P.L., J.Q.), Beijing, China 100191; and Key Laboratory of Assisted Reproduction (Y.Z., C.Z., Y.Y., R.L., N.L., J.Q.), Ministry of Education, Beijing, China 100191.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
authors.forename: Chunmei
authors.initials: C
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Nana
authors.initials: N
authors.lastname: Liu
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Qiao
mesh_terms: D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D018384:Oxidative Stress / Q000235:genetics*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011085:Polycystic Ovary Syndrome / Q000235:genetics / Q000378:metabolism*; D011446:Prospective Studies; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics*; D015854:Up-Regulation; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*; D000071818:Wnt-5a Protein
chemical_list: D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt