title: Progesterone modulates endothelial progenitor cell (EPC) viability through the CXCL12/CXCR4/PI3K/Akt signalling pathway.
abstract: OBJECTIVE
  Progesterone treatment can effectively increase levels of circulating endothelial progenitor cells (EPCs) and improve neurological functional outcome in a traumatic brain injury (TBI) rat model. However, the mechanisms of progesterone's effects on EPC viability remain elusive. The CXCL12/CXCR4 (CXC chemokine ligand 12/CXC chemokine receptor 4) signalling pathway regulates cell proliferation; we hypothesize that it mediates progesterone-induced EPC viability.
  
  
  METHODS
  EPCs were isolated from bone marrow-derived mononuclear cells (BM-MNCs) and treated with progesterone (5, 10 and 100 nm). MTS assay was used to investigate EPC viability. Protein expression was examined by Western blotting, ELISA assay and flow cytometry. Cell membrane and cytoplasm proteins were extracted with membrane and cytoplasm protein extraction kits. CXCR4 antagonist (AMD3100) and phosphatidylinositol 3-kinases (PI3K) antagonist (LY294002) were used to characterize underlying mechanisms.
  
  
  RESULTS
  Progesterone-induced EPC viability was time- and dose-dependent. Administration of progesterone facilitated EPC viability and increased expression of CXCL12 and phosphorylated Akt (also known as protein kinase B, pAkt) activity (P < 0.05). Progesterone did not regulate CXCR4 protein expression in cultured EPC membranes or cytoplasm. However, progesterone-induced EPC viability was significantly attenuated by AMD3100 or LY294002. Inhibition of the signalling pathway with AMD3100 and LY294002 subsequently reduced progesterone-induced CXCL12/CXCR4/PI3K/pAkt signalling activity.
  
  
  CONCLUSIONS
  The CXCL12/CXCR4/PI3K/pAkt signalling pathway increased progesterone-induced EPC viability.
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Yu
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Zhifei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Shengjie
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wen
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Quan
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Qilong
authors.initials: Q
authors.lastname: Tian
authors.affiliation: Department of Neurology, Henry Ford Hospital, Detroit, Michigan, 48202, USA.
authors.forename: Jieli
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Jianning
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, China.|Tianjin Neurological Institute, Tianjin, 300052, China.|Key Laboratory of Post-Neurotrauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, 300052, China.
authors.forename: Rongcai
authors.initials: R
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001596:Benzylamines; D048430:Cell Shape; D002470:Cell Survival / Q000187:drug effects; D054377:Chemokine CXCL12 / Q000378:metabolism*; D002867:Chromones / Q000494:pharmacology; D000080027:Cyclams; D066026:Endothelial Progenitor Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006571:Heterocyclic Compounds / Q000494:pharmacology; D008297:Male; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D017208:Rats, Wistar; D019718:Receptors, CXCR4 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors
chemical_list: D001596:Benzylamines; D054377:Chemokine CXCL12; D002867:Chromones; D000080027:Cyclams; D006571:Heterocyclic Compounds; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D011374:Progesterone; D051057:Proto-Oncogene Proteins c-akt; D019718:Receptors, CXCR4; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C088327:plerixafor
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