Yonsei Med J. 1998 Feb;39(1):1-12. English.
Published online Feb 20, 2002.
Copyright © 1998 The Yonsei University College of Medicine
Original Article

Hyperoxia influences mRNA expression of cytokines in cultured human umbilical vein endothelial cells

Min Soo Park and Heather M Wallace
    • Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea. yonsei.ac.kr
    • Department of Medicine and Therapeutics, University of Aberdeen, Aberdeen, UK.

Abstract

High concentrations of oxygen, indispensable for the treatment of severe hypoxemia from neonatal as well as adult respiratory distress syndrome, increase the risk of oxygen toxicity. Biochemical mechanisms are lipid peroxidation, protein sulfhydryl oxidation, enzyme inactivation, and DNA damage. Recent reports suggest that cytokines might be involved in free radical injury as well as in adaptive response to hyperoxic injury. However, actual signal transduction pathways involving cytokines have not yet been clarified. In this study we exposed cultured human umbilical vein endothelial cells (HUVECs) to either ambient air or 100% oxygen, and compared for the rate of DNA synthesis ([3H]thymidine uptake) at different time points up to 72 h. After exposing the cells to each treatment condition, we extracted RNA, constructed complementary DNA using reverse transcriptase, amplified the specific DNA segments of cytokines by polymerase chain reaction (PCR), and used the PCR products for gel electrophoresis to examine the bands which signified mRNA levels of corresponding cytokines. There was a significant decrease in the rate of DNA synthesis as early as 24 h. The mRNA expression of IL-1 β and TNFa seemed less influenced by hyperoxia, while IL-8 and TGF β showed marked increase in mRNA levels at 6 h of 100% oxygen exposure.

Keywords
Hyperoxia; reactive oxygen species; HUVEC; cytokine; RT-PCR


Metrics
Share
PERMALINK