Establishment of oxidative stress model with human valve interstitial cells mediated by h2o2
10.3724/SP.J.1008.2015.00001
- Author:
Pu-Xi XIONG
1
Author Information
1. Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
Cell model;
Human valve interstitial cells;
Hydrogen peroxide;
Oxidative stress
- From:
Academic Journal of Second Military Medical University
2015;36(1):1-5
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an oxidative stress model of human valve interstitial cells (VICs) mediated by hydrogen peroxide (H2 O2), so as to provide cytology model for research on pathogenesis of valvular heart diseases and screening anti-oxidant drugs. Methods The isolated and cultivated primary human VICs were divided into different groups randomly: control group containing DMEM culture medium with 10% FBS, experimental groups were treated with different concentrations (0, 50, 100, 300, 500, 800, and 1 000 μmol/L) of H2O2. H-E staining was used to observe cell morphology, MTT assay was used to estimate cell viability, and Annexin /PI flow cytometer was employed to evaluate the apoptosis of VICs. Results MTT assay showed that the survival rates of VICs were significantly different 24 h after exposure to different concentrations of H2O2(P<0.01), with those in 50 and 100 μmol/L groups being significantly higher than that of control group (P<0.05), and the cell survival rate began to decrease with the increase of H2O2 concentrations; the decrease became quicker when H2O2 concentrations was 800 μmol/L, with a survival rate of (69.8±8.3)%; and the survival rate decreased to (14.3±11.0)% when the concentration reached 1 000 μmol/L. H-E staining showed that at 800 μmol/L, H2O2 resulted in crimple and karyopyknosis of the VICs. Flow cytometer results confirmed apparent apoptosis of VICs at the concentration of 800 μmol/L, with the apoptosis already in the middle and advanced stages. Conclusion Oxidative stress model of VICs can be successfully established with H2O2, with the optimum concentration of H2O2 being 800 μmol/L and the expose period being 24 h.
