JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2015, Vol. 50 ›› Issue (09): 1-6.doi: 10.6040/j.issn.1671-9352.0.2015.197

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UVB damage and protection effect of ascorbic acid on human corneal endothelial cells in vitro

WEN Qian, FAN Ting-jun   

  1. Key Laboratory for Corneal Tissuse Engineering, Ocean University of China, Qingdao 266003, Shandong, China
  • Received:2015-04-27 Revised:2015-06-27 Online:2015-09-20 Published:2015-09-26

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Abstract: To postulate the damage effect of UVB and the protection effect of antioxidant ascorbic acid(Asc), the oxidantive effect of UVB, the protective effect of Asc, and the related molecular mechanisms were investigated systematically using an in vitro model of non-transfected human corneal endothelial(HCE) cells in this study. After HCE cells were exposed to UVB irradiation and/or Asc, the viability and morphology were examined by MTT assay and light microscopy, DNA oxidative damage was detected by 8-hydroxydeoxyguanosine immunofluorescent staining, and the level of reactive oxygen species(ROS) was assayed by dihydroethidium staining. Our results showed that 100~800 mj/cm2 UVB irradiation could decline the viability of HCE cells in a dose-and time-dependent manner. UVB at 200 mj/cm2(the average dose of UVB in natural sunlight) could induce cell shrinkage, oxidative damage of DNA, and elevation of ROS level significantly. Whereas, 1 mmol/L Asc, with significant enhancing effects on the viability and proliferation of HCE cells, could significantly reduce the oxidative damage of DNA and the ROS level in 200 mj/cm2 UVB-irradiated HCE cells. In conclusion, UVB irradiation has a significant damaging effect on HCE cells by inducing DNA oxidative damage via ROS generation, while Asc has an obvious protecting effect on UVB-irradiated HCE cells by eleminatingDNA oxidative damage via inhibiting ROS generation. These findings can be used as certain theory guidance in protecting HCE cells from UVB-induced oxidative damage by employing Asc and the other antioxidants.

Key words: oxidative damage, ascorbic acid, ultraviolet B, antioxidant, human corneal endothelial cell, reactive oxygen species

CLC Number: 

  • Q28
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