title: [Impact and related mechanisms of stromal cell-derived factor-1α on serum deprivation-induced cardiac stem cells apoptosis].
abstract: OBJECTIVE
  To explore the impact and related mechanisms of stromal cell-derived factor-1α (SDF-1α) on serum deprivation-induced apoptosis of cardiac stem cells (CSCs).
  
  
  METHODS
  CSCs were isolated from adult mouse heart tissue and cultured in vitro. Obtained cells were purified using magnetic-activated cell sorting (MACS) with c-kit magnetic beads. C-kit(+)CSCs were divided into five groups: normal control group, serum deprivation group, serum deprivation+SDF-1α group, serum deprivation+SDF-1α+AMD3100 group, serum deprivation+SDF-1α+LY294002 group. Cell apoptosis was assessed using the DeadEnd Colorimetric TUNEL System and flow cytometry analyses with an Annexin V-FITC Apoptosis Detection Kit. The viability of CSCs was assessed by CCK-8. The protein expression of Bcl-2 and phosphorylated Akt were detected by Western blot. The caspase-3 activity was determined using caspase-3 Colorimetric Assay Kit.
  
  
  RESULTS
  After magnetic separation, more than 85% of cardiosphere derived cells were positive for c-kit expression. Compared with the normal control group, the apoptosis rate of serum deprivation group was significantly increased[(27.03 ± 0.80)% vs. (1.51 ± 0.54)%, P < 0.01], which could be significantly reduced by SDF-1α in a concentration dependent manner and peak effect was seen with 100 ng/ml SDF-1α[(10.67 ± 1.06)% vs. (27.03 ± 0.80)%, P < 0.01]. The expressions of p-Akt and Bcl-2 were significantly increased and the activity of caspase-3 was significantly decreased in serum deprivation+SDF-1α group compared to serum deprivation group (P < 0.01). Further more, the expression of p-Akt and Bcl-2 were significantly decreased and the activity of caspase-3 was increased in both serum deprivation+SDF-1α+AMD3100 group and serum deprivation+SDF-1α+LY294002 group compared to serum deprivation+SDF-1α group (P < 0.01).
  
  
  CONCLUSIONS
  SDF-1α reduces serum deprivation induced CSCs apoptosis via modulating PI3K/Akt signaling pathway.
authors.affiliation: Department of Cardiology, Zhongda Hospital Affiliated Southeast University, Nanjing 210009, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.affiliation: Email: magenshan@hotmail.com.
authors.forename: Gen-shan
authors.initials: GS
authors.lastname: Ma
authors.forename: Xiao-dong
authors.initials: XD
authors.lastname: Pan
authors.forename: Zhong-pu
authors.initials: ZP
authors.lastname: Chen
authors.forename: Zu-long
authors.initials: ZL
authors.lastname: Sheng
authors.forename: Sheng-da
authors.initials: SD
authors.lastname: Hu
authors.forename: Yu-yu
authors.initials: YY
authors.lastname: Yao
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000494:pharmacology*; D003470:Culture Media / Q000737:chemistry; D051379:Mice; D009206:Myocardium / Q000166:cytology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D015398:Signal Transduction; D013234:Stem Cells / Q000187:drug effects*
chemical_list: D054377:Chemokine CXCL12; D003470:Culture Media; D019253:Proto-Oncogene Proteins c-bcl-2; C052617:Bcl2 protein, mouse; D051057:Proto-Oncogene Proteins c-akt; C505409:Casp3 protein, mouse; D053148:Caspase 3