title: [Hypoxia reduces the proliferation and occludin expression of primary sertoli cells].
abstract: OBJECTIVE
  To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli
  
  
  METHODS
  We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the Sertoli cells into five groups to be cultured in oxygen at the concentrations of 20%, 15%, 10%, 5% and 1%, respectively, for 6, 12, 24, 48 and 72 hours. We detected the proliferation of the Sertoli cells by CCK-8 assay, determined the expression of occludin by Western blot, and analyzed the differences among the five groups.
  
  
  RESULTS
  Oil red O staining revealed red lipid droplets in the cytoplasm of the Sertoli cells, and immunofluorescence showed the positive expression of the FasL protein, with the purity of Sertoli cells over 95% in vitro. Compared with the 20% normoxic group, the proliferation of the Sertoli cells was gradually reduced in the 15% and 10% hypoxia groups, and significantly declined in the 5% and 1% groups (P < 0.01). At 12 hours, the expression of occludin began to decrease with the prolonging of time and reduction of oxygen concentration (P < 0.01).
  
  
  CONCLUSIONS
  Hypoxia suppresses the proliferation of Sertoli cells and reduces the expression of occludin. It could be inferred that hypoxia could damage the integrity of blood-testis barrier and spermatogenesis of the testis.
authors.affiliation: Department of Urology, The Affiliated Hospital of Qingdao University School of Medicine, Qingdao, Shandong 266021, China.
authors.forename: Wei-Yu
authors.initials: WY
authors.lastname: Hao
authors.forename: Cui-Hua
authors.initials: CH
authors.lastname: Shao
authors.forename: You-Liang
authors.initials: YL
authors.lastname: Feng
authors.forename: Jian-Ting
authors.initials: JT
authors.lastname: Hu
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Hong-Qiang
authors.initials: HQ
authors.lastname: Wang
authors.forename: Pei-Tao
authors.initials: PT
authors.lastname: Wang
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002478:Cells, Cultured; D008297:Male; D062793:Occludin / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012708:Sertoli Cells / Q000378:metabolism*
chemical_list: D062793:Occludin