Involvement of heme oxygenase in PM2.5-toxicity in human umbilical vein endothelial cells
10.3760/cma.j.issn.0253-3758.2013.11.013
- VernacularTitle:血红素氧合酶-1在空气细颗粒物PM2.5致人脐静脉内皮细胞损伤过程中作用的实验研究
- Author:
Jing-Lu YANG
1
;
Ji-Yuan L(U)
;
Ming-Sheng ZHANG
;
Gang QIN
;
Cai-Ping LI
Author Information
1. 山西医科大学第一附属医院心内科
- Keywords:
Endothelial cells;
Reactive oxygen species;
Heme oxygenase-1;
PM2.5
- From:
Chinese Journal of Cardiology
2013;41(11):955-961
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the involvement of heme oxygenase (HO-1) in PM2.5 induced toxic responses in human umbilical vein endothelial cells (HUVECs).Methods The experiment groups are as follows:(1) control group; (2) PM2.5 groups:the cells were cultured with various concentrations of PM2.5 (200,400,800 μg/ml) for 24 h and 400 μg/ml was chosen for the main study; (3) PM2.5 + Trion group:the cells were pre-treated by 10 μmol/L Trion [a scavenger of reactive oxygen species(ROS)] for 1 h before PM2.5 (400 μg/ml) treatment for 24 h; (4) PM2.5 +ZnPP group:the cells were pretreated by HO-1 inhibitor ZnPP (10 μmol/L) for 1 h before treatment with PM2.5 (400 μg/ml) for 24 h.MTT assay was used to detect cell viability.Reverse transcription polymerase chain reaction (RT-PCR) and indirect immunofluorescence assay were used to determine the mRNA and protein expressions of HO-1.Fluorescence labeling probe method was used to measure intracellular ROS level and flow cytometry was used for cell apoptosis.Colorimetric assay was used to detect intracellular caspase-3 activity.Results Compared with control,PM2.5 significantly decreased cell viability,increased intracellular ROS,cell apoptosis and caspase-3 activity (all P ＜ 0.05),these effects were significantly attenuated in PM2.5 + Tiron group while enhanced in PM2.5 + ZnPP group (all P ＜ 0.05 vs.PM2.5 group).PM2.5 upregulated HO-1 mRNA and protein expressions in HUVECs which was downregulated in both PM2.5 + Tiron group and PM2.5 + ZnPP group.Conclusion PM2.5 could induce oxidative injury through increasing ROS production via modulating HO-1 mRNA and protein expressions,the injury could be aggravated with inhibition of the activity of HO-1 suggesting a potential protective role of HO-1 against PM2.5 induced oxidative stress in HUVECs.
