Effect of cigarette smoke extract and nicotine on the expression of thrombomodulin and endothelial protein C receptor in cultured human umbilical vein endothelial cells

Wei,Yujie; Lai,Bin; Liu,Huiliang; Li,Yi; Zhen,Wang; Fu,Ling

doi:10.3892/mmr.2017.8070

Effect of cigarette smoke extract and nicotine on the expression of thrombomodulin and endothelial protein C receptor in cultured human umbilical vein endothelial cells

Authors:
- Yujie Wei
- Bin Lai
- Huiliang Liu
- Yi Li
- Wang Zhen
- Ling Fu
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Affiliations: Institute of Cardiology, the General Hospital of Chinese People's Armed Police Forces, Beijing 100039, P.R. China, Department of Emergency, the General Hospital of Chinese People's Armed Police Forces, Beijing 100039, P.R. China, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, P.R. China
Published online on: November 15, 2017 https://doi.org/10.3892/mmr.2017.8070
Pages: 1724-1730
Copyright: © Wei et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study investigated the influence of cigarette smoke extract (CSE) and nicotine on the expression of thrombomodulin (TM) and endothelial protein C receptor (EPCR) in human umbilical vein endothelial cells (HUVECs). Smoking is associated with intravascular thrombosis. As a vital anticoagulation cofactor, TM is located on the endothelial cell surface and regulates intravascular coagulation by binding to thrombin, hence activating protein C. Activated protein C is a natural anticoagulant that interacts with EPCR to enhance the function of anticoagulation system. The effects of CSE (0.5‑5%) and nicotine (10‑3‑10‑9 mol/l) on the expression of TM and EPCR in HUVECs were observed. Reverse transcription‑quantitative polymerase chain reaction and flow cytometric analysis techniques were used for detecting TM and EPCR mRNA and protein expression levels, respectively. After 6‑h exposure, TM protein and mRNA expression levels decreased in a dose‑dependent manner. Stimulation with 5% CSE for 0, 6, 10, 12 and 24 h led to a decrease in the levels of TM mRNA and protein over time, which reached a peak at 12 h. The levels were significantly reduced compared with the control group (P<0.001). However, CSE had no effect on EPCR. Furthermore, nicotine had no influence on TM and EPCR. In conclusion, the present study supports a novel molecular mechanism of cigarette smoking‑associated thrombosis by the decreased expression of TM. Further studies are required to identify specific components in CSE responsible for decreasing TM expression and its associated consequences.

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