Open AccessProgesterone modulates endothelial progenitor cell ( EPC ) viability through the CXCL 12/ CXCR 4/ PI 3K/Akt signalling pathway
Author(s) -
Yu Peng,
Zhang Zhifei,
Li Shengjie,
Wen Xiaolong,
Quan Wei,
Tian Qilong,
Chen Jieli,
Zhang Jianning,
Jiang Rongcai
Publication year - 2016
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12231
Subject(s) - viability assay , cxc chemokine receptors , microbiology and biotechnology , chemokine , protein kinase b , biology , pi3k/akt/mtor pathway , progenitor cell , signal transduction , chemokine receptor , receptor , cell , stem cell , biochemistry
Objectives Progesterone treatment can effectively increase levels of circulating endothelial progenitor cells ( EPC s) 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 CXCL 12/ CXCR 4 ( CXC chemokine ligand 12/ CXC chemokine receptor 4) signalling pathway regulates cell proliferation; we hypothesize that it mediates progesterone‐induced EPC viability. Materials and methods EPC s were isolated from bone marrow‐derived mononuclear cells ( BM ‐ MNC s) and treated with progesterone (5, 10 and 100 n m ). 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. CXCR 4 antagonist ( AMD 3100) and phosphatidylinositol 3‐kinases ( PI 3K) antagonist ( LY 294002) 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 CXCL 12 and phosphorylated Akt (also known as protein kinase B, pA kt) activity ( P < 0.05). Progesterone did not regulate CXCR 4 protein expression in cultured EPC membranes or cytoplasm. However, progesterone‐induced EPC viability was significantly attenuated by AMD 3100 or LY 294002. Inhibition of the signalling pathway with AMD 3100 and LY 294002 subsequently reduced progesterone‐induced CXCL 12/ CXCR 4/ PI 3K/ pA kt signalling activity. Conclusions The CXCL 12/ CXCR 4/ PI 3K/ pA kt signalling pathway increased progesterone‐induced EPC viability.