title: Puerarin suppresses proliferation of endometriotic stromal cells partly via the MAPK signaling pathway induced by 17ß-estradiol-BSA.
abstract: BACKGROUND
  Puerarin is a major isoflavonoid compound extracted from Radix puerariae. It has a weak estrogenic action by binding to estrogen receptors (ERs). In our early clinical practice to treat endometriosis, a better therapeutic effect was achieved if the formula of traditional Chinese medicine included Radix puerariae. The genomic and non-genomic effects of puerarin were studied in our Lab. This study aims to investigate the ability of puerarin to bind competitively to ERs in human endometriotic stromal cells (ESCs), determine whether and how puerarin may influence phosphorylation of the non-genomic signaling pathway induced by 17ß-estradiol conjugated to BSA (E(2)-BSA).
  
  
  METHODS
  ESCs were successfully established. Binding of puerarin to ERs was assessed by a radioactive competitive binding assay in ESCs. Activation of the signaling pathway was screened by human phospho-kinase array, and was further confirmed by western blot. Cell proliferation was analyzed according to the protocol of CCK-8. The mRNA and protein levels of cyclin D1, Cox-2 and Cyp19 were determined by real-time PCR and western blotting. Inhibitor of MEK1/2 or ER antagonist was used to confirm the involved signal pathway.
  
  
  RESULTS
  Our data demonstrated that the total binding ability of puerarin to ERs on viable cells is around 1/3 that of 17ß-estradiol (E(2)). E(2)-BSA was able to trigger a rapid, non-genomic, membrane-mediated activation of ERK1/2 in ESCs and this phenomenon was associated with an increased proliferation of ESCs. Treating ESCs with puerarin abrogated the phosphorylation of ERK and significantly decreased cell proliferation, as well as related gene expression levels enhanced by E(2)-BSA.
  
  
  CONCLUSIONS
  Puerarin suppresses proliferation of ESCs induced by E(2)-BSA partly via impeding a rapid, non-genomic, membrane-initiated ERK pathway, and down-regulation of Cyclin D1, Cox-2 and Cyp19 are involved in the process. Our data further show that puerarin may be a new candidate to treat endometriosis.
authors.affiliation: Department of Traditional Chinese Medicine, Changhai Hospital and Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Cheng
authors.forename: Lizao
authors.initials: L
authors.lastname: Chen
authors.forename: Shengsheng
authors.initials: S
authors.lastname: Yang
authors.forename: Jie
authors.initials: J
authors.lastname: Han
authors.forename: Dongxia
authors.initials: D
authors.lastname: Zhai
authors.forename: Jian
authors.initials: J
authors.lastname: Ni
authors.forename: Chaoqin
authors.initials: C
authors.lastname: Yu
authors.forename: Zailong
authors.initials: Z
authors.lastname: Cai
mesh_terms: D000328:Adult; D001141:Aromatase / Q000235:genetics / Q000378:metabolism; D001667:Binding, Competitive; D049109:Cell Proliferation / Q000187:drug effects; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D004717:Endometrium / Q000187:drug effects* / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D008875:Middle Aged; D011485:Protein Binding / Q000187:drug effects; D011960:Receptors, Estrogen / Q000378:metabolism; D017154:Stromal Cells / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D007529:Isoflavones; D011960:Receptors, Estrogen; D019938:Cyclin D1; D004958:Estradiol; D001141:Aromatase; D051546:Cyclooxygenase 2; C033607:puerarin