Genoprotective effect of hyaluronic acid against benzalkonium chloride-induced DNA damage in human corneal epithelial cells.
OBJECTIVE
The aim of this study was to investigate hyaluronic acid (HA) protection on cultured human corneal epithelial cells (HCEs) against benzalkonium chloride (BAC)-induced DNA damage and intracellular reactive oxygen species (ROS) increase.


METHODS
Cells were incubated with different concentrations of BAC with or without the presence of 0.2% HA for 30 min. DNA damage to HCEs was examined by alkaline comet assay and by immunofluorescence microscopic detection of the phosphorylated form of histone variant H2AX (γH2AX) foci. ROS production was assessed by the fluorescent probe, 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Cell apoptosis was determined with annexin V staining by flow cytometry.


RESULTS
HA significantly reduced BAC-induced DNA damage as indicated by the tail length (TL) and tail moment (TM) of alkaline comet assay and by γH2AX foci formation, respectively. Moreover, HA significantly decreased BAC-induced ROS increase and cell apoptosis. However, exposure to HA alone did not produce any significant change in DNA damage, ROS generation, or cell apoptosis.


CONCLUSIONS
BAC could induce DNA damage and cell apoptosis in HCEs, probably through increasing oxidative stress. Furthermore, HA was an effective protective agent that had antioxidant properties and could decrease DNA damage and cell apoptosis induced by BAC.
Eye Center of the 2nd Affiliated Hospital, Medical College of Zhejiang University, Hangzhou, Zhejiang, China.
Han
H
Wu
Huina
H
Zhang
Changjun
C
Wang
Yihua
Y
Wu
Jiajun
J
Xie
Xiuming
X
Jin
Jun
J
Yang
Juan
J
Ye
D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001548:Benzalkonium Compounds / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D003315:Cornea / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019610:Cytoprotection / Q000502:physiology*; D004249:DNA Damage / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005452:Fluoresceins; D006657:Histones / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D018384:Oxidative Stress; D010766:Phosphorylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism; D013050:Spectrometry, Fluorescence
C110400:2',7'-dichlorodihydrofluorescein diacetate; D000975:Antioxidants; D001548:Benzalkonium Compounds; D005452:Fluoresceins; C468783:H2AX protein, human; D006657:Histones; D017382:Reactive Oxygen Species; D006820:Hyaluronic Acid