title: MiR 20a,-20b and -200c are involved in hydrogen sulfide stimulation of VEGF production in human placental trophoblasts.
abstract: Hydrogen sulfide (H2S) has been implicated to angiogenesis in various tissues. We sought to investigate the role of hydrogen sulfide (H2S) in regulating production of vascular endothelial growth factor (VEGF) proteins, the key factors of angiogenesis and vasculogenesis, in placenta.
  
  
  METHODS
  Placental tissues were obtained from pregnant women with preeclampsia and healthy pregnant women who underwent elective cesarean section. Explants and trophoblasts were isolated from healthy placentas and treated with H2S donor and precursor. Western blotting was used to determine the levels of cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE). The levels of VEGF mRNA, miR miR-200c,-20a and -20b were determined by quantitative real time PCR.
  
  
  RESULTS
  NaHS and l-cysteine increased VEGF but not placenta growth factor (PlGF) production in cultured explants and trophoblasts. Transfection of CBS and CSE siRNA reversed the stimulatory effect of l-cysteine on VEGF production in placental cells. H2S prolonged the half-life of VEGF mRNA and decreased the expression of miR-200c,-20a and -20b in placental cells. MiR-200c mimic and inhibitor affected VEGF mRNA and protein level, whereas miR-20a or -20b mimic and inhibitor affect VEGF protein release but not mRNA expression. The expression level of miR-200c,-20a and -20b as well as the level of CBS, CSE and VEGF were downregulated in preeclamptic placentas.
  
  
  CONCLUSIONS
  H2S produced via CSE and CBS plays a critical role in VEGF production in human placenta. Reduced expression of CSE and CBS may contribute to the abnormal production of angiogenic factors in preeclamptic placenta.
authors.affiliation: Department of Physiology, Second Military Medical University, Shanghai, China.
authors.forename: Tian-Xiao
authors.initials: TX
authors.lastname: Hu
authors.affiliation: Department of Physiology, Second Military Medical University, Shanghai, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: Maternity and Child Health Hospital of Pudong New District, Shanghai, China.
authors.forename: Xue-Jing
authors.initials: XJ
authors.lastname: Guo
authors.affiliation: Department of Obstetrics and Gynecology, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors.forename: Qian-Qian
authors.initials: QQ
authors.lastname: Sun
authors.affiliation: Department of Physiology, Second Military Medical University, Shanghai, China.
authors.forename: Ping
authors.initials: P
authors.lastname: He
authors.affiliation: Department of Obstetrics and Gynecology, Changhai Hospital, Second Military Medical University, Shanghai, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Gu
authors.affiliation: Maternity and Child Health Hospital of Pudong New District, Shanghai, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Physiology, Second Military Medical University, Shanghai, China. Electronic address: lu.gao@smmu.edu.cn.
authors.forename: Lu
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Physiology, Second Military Medical University, Shanghai, China. Electronic address: nixin@smmu.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Ni
mesh_terms: D002478:Cells, Cultured; D003541:Cystathionine beta-Synthase / Q000378:metabolism; D003542:Cystathionine gamma-Lyase / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism / Q000494:pharmacology*; D035683:MicroRNAs / Q000502:physiology*; D010920:Placenta / Q000187:drug effects / Q000378:metabolism; D011247:Pregnancy; D014327:Trophoblasts / Q000187:drug effects* / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: C533896:MIRN200 microRNA, human; C534451:MIRN20a microRNA, human; D035683:MicroRNAs; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003541:Cystathionine beta-Synthase; D003542:Cystathionine gamma-Lyase; D006862:Hydrogen Sulfide
keywords: Cystathionine β-synthase; Cystathionine γ-lyase; Placenta; Vascular endothelial growth factor; miRNAs