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The protective effects of 6‐ C y S e CD with GP x activity against UVB ‐induced injury in H a C a T cells
Author(s) -
Li Xin,
Zhang Lin,
Xu YaWei,
Wang Cheng,
Zhao Yang,
Yu Ping,
Lv ShaoWu,
Yan GangLin,
Liu JunQiu,
Luo GuiMin
Publication year - 2013
Publication title -
australasian journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.67
H-Index - 53
eISSN - 1440-0960
pISSN - 0004-8380
DOI - 10.1111/j.1440-0960.2012.00962.x
Subject(s) - hacat , microbiology and biotechnology , viability assay , apoptosis , lipid peroxidation , reactive oxygen species , dna damage , cell cycle , flow cytometry , oxidative stress , glutathione peroxidase , dna fragmentation , keratinocyte , chemistry , biochemistry , programmed cell death , biology , superoxide dismutase , dna , in vitro
Background The generation of harmful reactive oxygen species ( ROS ) induced by UVB irradiation could induce cell apoptosis and change the cell cycle. 6A , 6A ′‐dicyclohexylamine‐ 6B , 6B ′‐diselenide‐bis‐β‐cyclodextrin (6‐ C y S e CD ) is a novel glutathione peroxidase ( GPx ; EC 1.11.1.9) mimic. The aim of this study was to investigate the anti‐oxidative effects of 6‐ C y S e CD in cultured immortalised human keratinocyte cells ( H a C a T ). Methods H a C a T cells were treated with 30 mJ /cm 2 UVB to establish a damage model. The cultured H a C a T cells were randomly assigned to the control, UVB and treatment groups. The treatment group was incubated with 20 μmol/L of GPx mimics before UVB irradiation. Cell viability was detected by (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay, the level of lipid peroxidation was determined by the formation of malondialdehyde ( MDA ), DNA fragmentation was observed using agarose gel electrophoresis and the levels of intracellular ROS and cell cycle progression were measured by flow cytometry. Results The levels of cytotoxicity, intracellular ROS , lipid peroxidation and oxidative DNA damage significantly increased after UVB irradiation in the H a C a T cells. UVB irradiation caused pre‐ G 1 ‐phase arrest in H a C a T cells and significantly reduced the number of HaCaT cells in the S phase. The GPx mimics 6‐ C y S e CD and 2‐phenyl‐l,2‐benzisoselenazol‐3( 2H )‐one (ebselen) significantly blocked UVB ‐induced apoptosis and changed the cell cycle of the H a C a T cells. The blocked effect of pretreatment 6‐ C y S e CD in UVB ‐irradiated H a C a T cells was better than that of pretreatment with ebselen. Conclusion 6‐ C y S e CD can relieve the damage induced by UVB irradiation in H a C a T cells.