南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (10): 1434-.

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三黄茵赤方含药血清对LO2细胞DNA氧化损伤的影响

代欢,刁建新,区锦莹,李海叶,杨运高   

  • 出版日期:2015-10-20 发布日期:2015-10-20

Protective effect of Sanhuangyinchi Fang drug serum on hydrogen peroxide-induced
DNA oxidative damage in LO2 cells

  • Online:2015-10-20 Published:2015-10-20

摘要: 目的探讨三黄茵赤方含药血清对H2O2诱导LO2 细胞DNA氧化损伤的保护作用。方法采集三黄茵赤方含药血清,以
LO2细胞株为研究对象,通过H2O2 诱导LO2细胞DNA氧化损伤模型,实验分正常对照组、H2O2模型组、三黄茵赤方含药血清
5%、10%、15%浓度组、维生素E组,采用倒置显微镜观察各组形态,CCK-8法测定细胞存活率,生化法检测细胞内SOD、CAT和
GSH-PX活性,流式细胞仪检测细胞内ROS水平,ELISA检测细胞内8-OHdG含量,彗星实验检测细胞DNA损伤情况。结果
与H2O2模型组比较,三黄茵赤方含药血清10%、15%浓度组、维生素E组均能提高细胞存活率(P<0.05),增强细胞内SOD、CAT、
GSH-PX活性(P<0.05),改善细胞对ROS清除能力(P<0.01),降低细胞内8-OHdG含量(P<0.01),减低细胞拖尾率、尾长、尾矩及
Olive尾矩(P<0.05)。结论三黄茵赤方含药血清对H2O2诱导LO2细胞DNA氧化性损伤具有保护作用,以15%浓度含药血清组
作用最显著。

Abstract: Objective To study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen
peroxide-mediated DNA oxidative damage in LO2 cells. Methods The LO2 cells were randomly divided into the control
group , H2O2 group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were
observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of
SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA
damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively. Results Compared with H2O2
group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT,
GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced
tailing ratio, tail length, tail moment and Olive tail moment (P<0.05). Conclusion SF drug serum, especially at the
concentration of 15%, can protect LO2 cells from H2O2-mediated DNAoxidative damage.