title: Glucose-induced increase in circulating progenitor cells is blunted in polycystic amenorrhoeic subjects.
abstract: BACKGROUND
  Glucose-induced kinetics of bone marrow-derived stem cells in healthy females is presently unknown. The objectives of this study were to determine whether circulating levels of CD133(+), CD34(+) and CD133(+)CD34(+) cells increase in response to glucose load in healthy females and whether the kinetics is altered in amenorrhoeic women. The other objective of the work was to compare the endothelial differentiation potential of peripheral blood-derived endothelial progenitor cells (EPCs) from healthy versus amenorrhoeic women.
  
  
  METHODS
  In this case-control study, 44 amenorrhoeic subjects and 36 age-matched females with no menstrual disturbance were recruited at Apollo Hospitals, a Tertiary health care center in Chennai, India. Circulating bone marrow-derived stem cells were measured by two color direct flow cytometry. Cultured progenitor cells were characterized at Day 7 and 14 for expression of endothelial markers and production of nitric oxide (NO) via immunofluoroscence.
  
  
  RESULTS
  The amenorrhoeic subjects were insulin resistant with homeostatic model of assessment of insulin resistance values of 3.33 ± 0.3 versus 1.75 ± 0.148 observed for controls (P< 0.0001). Among the amenorrhoeic subjects, 38 subjects had polycystic ovaries with no signs of hyperandrogenism. Fasting levels of CD133(+), CD34(+) and CD133(+)CD34(+) cells were reduced in amenorrhoeic subjects (P< 0.001). There was a 1.5 to 2-fold increase in the circulating levels of these cells in response to 75 g oral glucose challenge at 1 and 2 h post-load conditions in controls, which was significantly blunted for CD133(+) (P< 0.001) and CD133(+)CD34(+) (P< 0.001) cells in amenorrhoeic subjects. A positive correlation was observed between estrogen and fasting CD133(+) (r= 0.205, P= 0.070), CD34(+) (r= 0.249, P= 0.027) and CD133(+)CD34(+) (r= 0.217, P= 0.055) cell counts. Additionally, fasting counts for CD34(+) and CD133(+)CD34(+) cells positively correlated with FSH and inversely correlated with LH and C-peptide in the polycystic group. Cultured cells from polycystic subjects exhibited reduced adherence to fibronectin and expressed lower levels of endothelial nitric-oxide synthase and NO.
  
  
  CONCLUSIONS
  Oral glucose-induced increase in circulating numbers of CD133(+) and CD133(+)CD34(+) cells and endothelial differentiation potential of peripheral blood-derived EPCs is attenuated in insulin resistant amenorrhoeic subjects.
authors.affiliation: Laboratory of Vascular Biology, Department of Biotechnology, Indian Institute of Technology Madras, BT 415, Chennai 600036, India.
authors.forename: Soumi
authors.initials: S
authors.lastname: Bairagi
authors.forename: Jayashree
authors.initials: J
authors.lastname: Gopal
authors.forename: Abel A
authors.initials: AA
authors.lastname: Nathan
authors.forename: Subash S
authors.initials: SS
authors.lastname: Babu
authors.forename: N Pavan
authors.initials: NP
authors.lastname: Kumar
authors.forename: Madhulika
authors.initials: M
authors.lastname: Dixit
mesh_terms: D000071916:AC133 Antigen; D000568:Amenorrhea / Q000097:blood / Q000150:complications / Q000473:pathology*; D015703:Antigens, CD / Q000097:blood; D016022:Case-Control Studies; D004967:Estrogens / Q000097:blood; D005215:Fasting; D005260:Female; D005434:Flow Cytometry; D051157:Frizzled Receptors / Q000097:blood; D005947:Glucose / Q000494:pharmacology*; D005951:Glucose Tolerance Test; D006023:Glycoproteins / Q000097:blood; D006801:Humans; D007333:Insulin Resistance; D010455:Peptides / Q000097:blood; D011085:Polycystic Ovary Syndrome / Q000097:blood / Q000150:complications / Q000473:pathology*; D013234:Stem Cells / Q000187:drug effects*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D004967:Estrogens; C549560:FZD9 protein, human; D051157:Frizzled Receptors; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D005947:Glucose