[Advanced oxidation protein products promote expression of stromal-cell derived factor-1alpha of ECV304 cells through ERK signal pathway]
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Published on: July 20, 2014
Chun-Hong Shi1, Yi-Nong Jiang, Lu-Juan Shan
1Department of Endocrinology, Dalian Medical University, Dalian 116011.
Objective:
To explore the effects of advanced oxidation protein products (AOPP) on expressions of stromal cell-derived factor-1alpha (SDF-1alpha) in ECV304 cells and the signal pathway that mediated the effects.
Methods:
AOPP-BSA was made from bovine serum albumin (BSA) and sodium hypochlorite. After treated with AOPP-BSA of different concentrations (50, 100, 200 micromol/L), the expressions of SDF-1alpha mRNA in ECV304 cells were measured by reverse transcription-polymerase chain reaction (RT-PCR) and the expressions of SDF-1alpha protein and the levels of phosphorylated extracellular signal-regulated kinase (ERK) in ECV304 cells were analyzed by Western blot. In inhibition test, U0126, the special inhibitor of ERK of different concentrations (0.1, 1, 10 rmol/L) were added into ECV304 cells culture media for 1 hour, then the cells were treated with AOPP-BSA for 24 hours, at last the protein levels in supernatant were detected by enzyme-linked immunosorbent assay (ELISA).
Results:
AOPP-BSA obviously promoted the expressions of SDF-1alpha mRNA and increased the levels of SDF-1beta protein of ECV304 cells in dose-dependent manner (all P < 0.01), after 15 minutes treated with 200 micromol/L AOPP-BSA, the levels of phosphorylated ERK of ECV304 cells increased significantly (P < 0.01). When the ERK pathway was blocked by U0126, the promoting effects of AOPP-BSA on expressions of SDF-la protein in ECV304 cells were significantly inhibited in dose-dependent manner (P < 0.05).
Conclusion:
AOPP induced the expression of SDF-la of ECV304 cells, ERK signal pathway is an important pathway that mediated the effects.
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