Hepatocyte growth factor stimulates proliferation and migration during wound healing of retinal pigment epithelial cells in vitro.
OBJECTIVE
A defect in retinal pigment epithelial (RPE) cells may cause dysfunction of the neural retina, so rapid recovery of differentiated RPE cells is required after RPE injury. We investigated the effect of hepatocyte growth factor (HGF) on wound healing in RPE cells.


METHODS
Confluent monolayers of bovine RPE cells were denuded, and the cells were allowed to recover in the presence or absence of HGF. The effect of HGF on RPE cell proliferation was evaluated by a 3-(4;5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetraz olium assay. In a migration assay, mitomycin C was used to inhibit proliferation, and the number of migrated cells was counted. The signaling pathways involved were examined using inhibitors of mitogen-activated protein kinase (MAPK), phosphatidylinositol-3 (PI3) kinase and protein kinase C pathways.


RESULTS
At 80 ng/mL, HGF stimulated the wound closure of RPE monolayers and rendered the restituted cells more epithelioid in shape. HGF at 10 ng/mL stimulated RPE cell migration the most, whereas 80 ng/mL of HGF inhibited migration, but stimulated proliferation the most. In particular, PI3 kinase and MAPK inhibitor inhibited PRE cell migration and proliferation, respectively.


CONCLUSIONS
HGF stimulated wound closure in cultured RPE cells, and rendered restituted cells epithelioid in shape. HGF may become a therapeutic candidate for RPE wound healing.
Miura
Yoko
Y
Department of Ophthalmology and Visual Sciences, Osaka City University Graduate School of Medicine, Osaka, Japan.
Yanagihara
Nobuyo
N
Imamura
Hitoshi
H
Kaida
Mayumi
M
Moriwaki
Mitsuyasu
M
Shiraki
Kunihiko
K
Miki
Tokuhiko
T
D000818:Animals; D002417:Cattle; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D015398:Signal Transduction; D014945:Wound Healing / Q000187:drug effects*
D004791:Enzyme Inhibitors; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017228:Hepatocyte Growth Factor; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases