[Oxidative stress experimental model of rat with stria vascularis marginal cells injury induced by hydrogen peroxide in vitro].
OBJECTIVE
To set up the oxidative stress experimental model of rat cochlea with stria vascularis marginal cells injury induced by hydrogen peroxide in vitro.


METHODS
Cultured marginal cells of rat were treated by 200, 300, 400, 600 and 800 micromol/L hydrogen peroxide (H(2)O(2)) for 0.5, 1, 2, 4, 16 and 24 hours, respectively. Cell viability was assessed by the CCK-8 assay. The content of the lipid peroxidation production malondialdehyde (MDA) were detected in H(2)O(2) induced marginal cells injury with different concentration H(2)O(2). Apoptosis was assessed by flow cytometry by propidium sodium staining. The expression of the cleaved-caspase-3 was assessed by Western blot.


RESULTS
Being exposed to H(2)O(2), marginal cells displayed nuclear pyknosis and margination, cytoplasmic condensation, cell shrinkage and formation of membrane and bounded apoptotic bodies. A time-dependent and dose-dependent decrease of cellular viability was detected with the treatment of H(2)O(2). Cellular maleic dialdehyde was generated in proportion to the concentration of H(2)O(2) at 2 hours and the number of apoptotic cells increased significantly (P < 0. 05). Western blot showed the expression of the cleaved-caspase-3 increased when 200 micromol/L, 300 micromol/L and 400 micromol/L H(2)O(2) treated cultured marginal cells. Thereafter the expression of the cleaved-caspase-3 decreased with 600 micromol/L H(2)O(2) and with 800 micromol/L H(2)O(2) the expression of cleaved-caspase-3 was weak.


CONCLUSIONS
The findings indicated that the experimental model can be established successfully using cultured cells exposed to H(2)O(2) and activation of caspase-3 is associated with hydrogen peroxide induced rat marginal cells the oxidative stress injury.
Li
Jun
J
Department of Otorhinolaryngology, Wuhan Union Hospital, Tongji Medical College of Huazhong Universty of Science and Technology, China.
Kong
Wei-jia
WJ
Zhao
Xue-yan
XY
Hu
Yu-juan
YJ
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D004195:Disease Models, Animal; D006861:Hydrogen Peroxide / Q000378:metabolism*; D015227:Lipid Peroxidation; D018384:Oxidative Stress; D051381:Rats; D017208:Rats, Wistar; D013316:Stria Vascularis / Q000378:metabolism*; D014012:Tinnitus / Q000378:metabolism*
D006861:Hydrogen Peroxide; C505410:Casp3 protein, rat; D053148:Caspase 3