Research On The Antagonistic Effects Of Zinc Sulfate Against Oxidative Damage Caused By Ultraviolet Radiation

ROS and free radical have been playing essential roles on skin damage which caused by ultraviolet irradiated, such as on photoaging and even on the procedure of carcinogenesis. The skin acute damage which caused by ultraviolet exposed and the skin chronic damage which caused by UVB exposed could be prevented by supplied antioxidant which can eliminate free radical.Object: To discuss the expression of the inducer of zinc sulfate, and the antagonism effects and mechanism of the inducer to skin oxidative damage which caused by UVB exposure. To discuss the methods of disparted and cultured of keratinocytes. Method: To used methods of dispase combined with parenzyme to obtain mononuclearcell from back skins of newborn neonate rats. It was used adherence culture combined with method of digested time control to obtain high pure keratinocytes (KC). Then the keratinocytes was cultured with L-DMEM(contained 10%FBS)and F12 for amplification. When cell fusing rate reached 90%, it was added zinc sulfate for induction culture. The cells irradiated by UVB when the inducer of zinc sulfate expressed, then observed affects of the inducer to the effect of UVB in the situation of cell culture in vitro. The protein expression of inducer was determined by polyacrylamide gel electrophoresis.Result:1. It was obtained high pure adherence KC. The primary culture showed KC appeared road stone and had well-distributed. The homogeneity of KC obviously improved during subculturing. There was no morphology change after serial subculturing over 3 generation and KC was stable.2. When the inducer of 0.0001% zinc sulfate expressed after suuculturing over 3 generation, experiment group and control group were irradiated by UVB. Then, antioxidase (SOD, GSH and GSH-Px) and oxidant damage index (MDA, hydroxyl radical and superoxide anion) were determined. The differences between two groups were significate (P<0.05). The result showed: the concentrations of MDA and free radical in KC, which in the system of cell cultured in vitro that contained zinc sulfate, was obviously reduced. The concentration of hydroxyl radical of experiment group is 23.926±5.804, and that of control group is 63.643±4.723, the differences between groups was significant (P<0.05). And the concentration of MDA and superoxide anion had the same trend. The concentrations of SOD, GSH and the activities of GSH-Px in KC of experiment group were significant higher than that of the control group.Conclusion:1. It is considered to be convenient, efficient and useful method to used dispase combined with parenzyme, adherence culture and tissues culture under the strict control of digested time for obtaining KC which purity and activity are both high, after departed and cultured of KC of neonate rat, and observed and determined morphology and biology features of KC.2. the KCs inducted by zinc sulfate have the ability of protective and the antagonistic effects against oxidabive damage caused by UVB irradiation .The mechanism of those effects still needs to further investigate.