Hepatocyte growth factor protects against apoptosis induced by advanced glycation end products in endothelial cells.
OBJECTIVE
To investigate the effects of hepatocyte growth factor (HGF) on vascular endothelial cells apoptosis induced by advanced glycation end products (AGEs) and its possible mechanism.


METHODS
Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and intervened by different concentrations of AGEs and HGF. The cell inhibitory rates of each group with different culture time (12, 24, 48, and 72 hours) were measured by methyl thiazolyl tetrazolium (MTT) assay. The early stage apoptosis was detected by flow cytometry with Annexin V-FITC/PI double staining, morphology of cell apoptosis was observed by hoechst 33258 fluorescence staining, and the expression of apoptosis-associated genes Bax and Bcl-2 were determined by Western blotting. The activity of caspase-3 was detected by enzyme-linked immunosorbent assay (ELISA).


RESULTS
Morphological observation indicated that high concentration of AGEs induced characteristic apoptotic changes in HUVECs. Within a certain concentration range, HUVECs apoptosis inducing rates by AGEs were in both dose- and time-dependent manners. HGF significantly inhibited the apoptosis of HUVECs induced by AGEs (P < 0.05). AGEs significantly promoted expression of Bax protein, but not Bcl-2. Whereas HGF significantly promoted the expression of Bcl-2 (P < 0.01) and decreased the activity of caspase-3 (P < 0.05) without affecting Bax level.


CONCLUSIONS
AGEs can induce the apoptosis of endothelial cells in vitro. HGF may effectively attenuate AGEs-induced endothelial cells apoptosis through upregulating Bcl-2 gene expression and inhibiting caspase-3 activation.
Zhou
Yi-jun
YJ
Department of Endocrinology and Metabolism, the First Affiliated Hospital, China Medical University, Shenyang 110001.
Wang
Jia-he
JH
Zhang
Jin
J
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D017127:Glycation End Products, Advanced; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D017228:Hepatocyte Growth Factor; D053148:Caspase 3