title: Attenuation of glomerular endothelial cells from high glucose-induced injury by blockade of MAD2B.
abstract: OBJECTIVE
  To assess the role of mitotic arrest-deficient 2-like protein 2 (MAD2B) in high glucose-induced injury in mouse glomerular endothelial cells (GEnCs).
  
  
  METHODS
  GEnCs were cultured in vitro, and MAD2B protein levels were measured by Western blot in cells stimulated with high glucose (30 mM) for various periods of time. MAD2B and scrambled shRNA were introduced into GEnCs by liposomal transfection. Cell proliferation, apoptosis, nitric oxide (NO) production, and monolayer permeability were then measured in cells grown in the following conditions: control, high glucose treatment, MAD2B shRNA transfection with high glucose treatment, and scrambled shRNA transfection with high glucose treatment.
  
  
  RESULTS
  High glucose increased the protein levels of MAD2B in GEnCs. Compared with control cells, apoptosis was increased by high glucose treatment, which was attenuated by transfection with MAD2B shRNA transfection. Cells treated with high glucose produced less NO than control cells, whereas MAD2B shRNA transfection increased NO production. Cell monolayer permeability was enhanced in high glucose treated cells, but MAD2B shRNA transfection reduced permeability.
  
  
  CONCLUSIONS
  High glucose levels induced the expression of MAD2B in GEnCs, whereas suppressing its expression reduced high glucose-induced endothelial cell apoptosis and high permeability, and promoted cell proliferation and NO production.
authors.affiliation: Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Yu-Mei
authors.initials: YM
authors.lastname: Wang
authors.forename: Yu
authors.initials: Y
authors.lastname: Hao
authors.forename: Xian-Fang
authors.initials: XF
authors.lastname: Meng
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: He
authors.forename: Shan
authors.initials: S
authors.lastname: Chen
authors.forename: Pan
authors.initials: P
authors.lastname: Gao
authors.forename: Hui
authors.initials: H
authors.lastname: Tang
authors.forename: Hua
authors.initials: H
authors.lastname: Su
authors.forename: Chun
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology*; D064246:Mad2 Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D064246:Mad2 Proteins; C577699:Mad2l2 protein, mouse; D034741:RNA, Small Interfering; D009569:Nitric Oxide; D005947:Glucose