Protective effect of paeoniflorin against oxidative stress in human retinal pigment epithelium in vitro.
OBJECTIVE
This study was conducted to determine whether paeoniflorin (PF) could prevent H₂O₂-induced oxidative stress in ARPE-19 cells and to elucidate the molecular pathways involved in this protection.


METHODS
Cultured ARPE-19 cells were subjected to oxidative stress with H₂O₂ in the presence and absence of PF. The preventive effective of PF on reactive oxygen species (ROS) production and retinal pigment epithelium (RPE) cell death induced by H₂O₂ was determined by 2',7'- dichlorodihydroﬂuorescein diacetate (H₂DCFDA) fluorescence and 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide (MTT) assay. The ability of PF to protect RPE cells against ROS-mediated apoptosis was assessed by caspase-3 activity and 4', 6-diamidino-2-phenylindole (DAPI) staining. Furthermore, the protective effect of PF via the mitogen-activated protein kinase (MAPK) pathway was determined by western blot analysis.


RESULTS
PF protected ARPE-19 cells from H₂O₂-induced cell death with low toxicity. H₂O₂-induced oxidative stress increased ROS production and caspase-3 activity, which was significantly inhibited by PF in a dose-dependent manner. Pretreatment with PF attenuated H₂O₂-induced p38MAPK and extracellular signal regulated kinase (ERK) phosphorylation in human RPE cells, which contributed to cell viability in ARPE-19 cells.


CONCLUSIONS
This is the first report to show that PF can protect ARPE-19 cells from the cellular apoptosis induced by oxidative stress. The results of this study open new avenues for the use of PF in treatment of ocular diseases, such as age-related macular degeneration (AMD), where oxidative stress plays a major role in disease pathogenesis.
Department of Ophthalmology, People's Eye Institute, Peking University People's Hospital, Key Laboratory of Vision Loss and Restoration, Ministry of Education, Beijing, China.
Xie
X
Wankun
Yu
Y
Wenzhen
Zhao
Z
Min
Zhou
Z
Weiyan
Chen
C
Huan
Du
D
Wei
Huang
H
Lvzhen
Yongsheng
Y
Xu
Li
L
Xiaoxin
D017209:Apoptosis / Q000187:drug effects; D001565:Benzoates / Q000494:pharmacology*; D001952:Bridged-Ring Compounds / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005960:Glucosides / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D039821:Monoterpenes; D018384:Oxidative Stress / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D006027:Glycosides
D001565:Benzoates; D001952:Bridged-Ring Compounds; D053148:Caspase 3; D048049:Extracellular Signal-Regulated MAP Kinases; D005960:Glucosides; D006861:Hydrogen Peroxide; D039821:Monoterpenes; D017382:Reactive Oxygen Species; D048051:p38 Mitogen-Activated Protein Kinases; C015423:peoniflorin; D006027:Glycosides