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Native low density lipoprotein increases the production of both nitric oxide and reactive oxygen species in the human umbilical vein endothelial cells

Genes & Genomics volume 41, pages 373–379 (2019)Cite this article

Abstract

Background

Nitric oxide synthases (NOSs) are a unique family of enzymes that catalyze the production of nitric oxide (NO) from l-arginine. Atherogenic action of oxidized low-density lipoproteins (oxLDL) may be mediated partly by the formation of NO in endothelial cells.

Objective

The objective of this study was to identify sources of reactive oxygen species (ROS) causing native LDL (nLDL)-induced senescence of cultured human umbilical vein endothelial cells (HUVECs).

Methods

HUVECs were treated with nLDL and NO production was assessed using Griess reagent as substrate and spectrophotometry in the absence or presence of specific inhibitors of endothelial NOS (eNOS) and inducible NOS (iNOS). In addition, expression levels of eNOS and iNOS were measured with ELISA and western blotting, and ROS was evaluated using 2′,7′-dichlorofluorescin diacetate (DCF-DA) and a fluorescence microplate reader.

Results

NO formation in nLDL-treated HUVECs was significantly increased. Long-term treatment with nLDL up-regulated both eNOS and iNOS proteins. Such increase of NO production in HUVECs induced by nLDL was significantly suppressed by treatment with iNOS-selective inhibitor 1400 W, but not by the eNOS-selective inhibitor L-NIO. Native LDL treatment uncoupled Hsp90, the regulatory binding protein of eNOS, from the enzyme in HUVECs. Native LDL also significantly increased ROS production in HUVECs.

Conclusion

These findings suggest that oxidative stress originated from induction of iNOS and eNOS could be a causative factor for nLDL-induced senescence of HUVECs.

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Author notes

  1. Kee Oh Chay and Sung Yeul Yang have contributed equally to this work.

Affiliations

  1. Department of Biochemistry, Medical School, The Research Institute of Medical Science, Chonnam National University, 264 Seoyang-ro, Hwasun-eup, Hwasun-gun, Jeollanam-do, 58128, South Korea

    Hyun Joong Yoon, Kee Oh Chay & Sung Yeul Yang

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  1. Hyun Joong Yoon
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  2. Kee Oh Chay
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Correspondence to Sung Yeul Yang.

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Yoon, H. J., Chay, K.O., and Yang, S.Y. declares that they have no conflicts of interest.

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Yoon, H.J., Chay, K.O. & Yang, S.Y. Native low density lipoprotein increases the production of both nitric oxide and reactive oxygen species in the human umbilical vein endothelial cells. Genes Genom 41, 373–379 (2019). https://doi.org/10.1007/s13258-018-00777-4

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Keywords

  • Native low density lipoprotein
  • Human umbilical vein endothelial cell
  • Nitric oxide
  • Reactive oxygen species
  • Inducible nitric oxide synthase