Protective Role of Hinokitiol Against H2O2-Induced Injury in Human Corneal Epithelium.
We recently found that hinokitiol has anti-inflammatory activity in human corneal epithelial (HCE) cells. Herein, we investigated the protective role of hinokitiol against H2O2-induced injury in HCE cells and the mechanisms that underlie its action.



HCE cells were incubated with different concentrations of hinokitiol or dimethylsulfoxide (DMSO), which served as a vehicle control, before H2O2stimulus. The cell viability was evaluated using a cell counting kit-8 (CCK-8) assay. TUNEL, phosphorylated histone γH2A.X, cleaved caspase-3 expression analyses, and location of cytochrome c were conducted to detect cell injury and apoptosis. Reactive oxygen species (ROS), catalase (CAT), superoxide dismutase (SOD), methane dicarboxylic aldehyde (MDA), and total antioxidative capacity (T-AOC) were used to determine oxidative stress. Bcl-2 and Bax protein expressions were measured by western blotting.



Hinokitiol significantly improved the cell viability, decreased the apoptosis rate, inhibited DNA damage, and reduced cleaved caspase-3 expression and the leakage of cytochrome c from mimitochondrion to cytoplasm of HCE cells against the oxidative stress induced by H2O2. Generation of ROS and MDA and decreased activity of CAT, SOD, and T-AOC were also ameliorated by hinokitiol administration. Moreover, Bcl-2 expression was down-regulated while Bax was up-regulated by H2O2stimulus, which were reversed by hinokitiol application.



Hinokitiol protects HCE cells against H2O2-induced injury likely by its antioxidant activity and modulating the Bcl-2 signaling pathway.
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Yufeng
Y
Xu
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Shengzhan
S
Wang
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Qi
Q
Miao
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Kai
K
Jin
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Lixia
L
Lou
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Xin
X
Ye
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Yan
Y
Xi
a Department of Ophthalmology , The Second Affiliated Hospital of Zhejiang University, College of Medicine , Hangzhou , Zhejiang , China.
Juan
J
Ye
D000890:Anti-Infective Agents / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D019610:Cytoprotection; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D020287:In Situ Nick-End Labeling; D039821:Monoterpenes / Q000494:pharmacology*; D016877:Oxidants / Q000633:toxicity*; D018384:Oxidative Stress; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D014334:Tropolone / Q000031:analogs & derivatives* / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D000890:Anti-Infective Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; C494826:BAX protein, human; D039821:Monoterpenes; D016877:Oxidants; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D014334:Tropolone; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D053148:Caspase 3; C009479:beta-thujaplicin; D004121:Dimethyl Sulfoxide
Antioxidant; ROS; apoptosis; hinokitiol; human corneal epithelial cells