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Natural and synthetic flavonoids, novel blockers of the volume-regulated anion channels, inhibit endothelial cell proliferation

  • Ion channels, receptors and transporters
  • Published:
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Abstract

Natural flavonoids are ubiquitous in dietary plants and vegetables and have been proposed to have antiviral, antioxidant, cardiovascular protective, and anticancer effects. Volume-regulated anion channels (VRACs), which are essential for cell volume regulation, have been proposed to play a key role in cell proliferation and migration, apoptosis, transepithelial transport, and cancer development. In this study, we screened a group of 53 structurally related natural flavonoids and three synthetic flavonoids for their inhibitory activities on VRAC currents. A whole-cell patch technique was used to record VRAC currents in the human embryonic kidney (HEK) 293 and human umbilical vein endothelial (HUVEC) cells. The 5′-bromo-2-deoxyuridine (BrdU) assay technique was used to investigate cell proliferation. At 100 μM, 34 of 53 compounds significantly inhibited hypotonic extrasolution-induced VRAC currents by > 50% in HEK293 cells. Among these compounds, luteolin, baicalein, eupatorin, galangin, quercetin, fisetin, karanjin, Dh-morin, genistein, irisolidone, and prunetin exhibited the highest efficacy for VRAC blockade (the mean inhibition > 80%) with IC50s of 5–13 μM and Emaxs of about 87–99%. We also studied the effects of three synthetic flavonoids on VRAC currents in HEK293 cells. Flavoxate showed high inhibition efficacy toward VRAC currents (IC50 = 2.3 ± 0.3 μM; Emax = 91.8% ± 2.7%). Finally, these flavonoids inhibited endogenous VRAC currents and cell proliferation in endothelial cells. This study demonstrates that natural and synthetic flavonoids are potent VRAC current inhibitors, and VRAC inhibition by flavonoids might be responsible for their anti-angiogenic effects.

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Abbreviations

VRACs:

volume-regulated anion channels

HEK:

human embryonic kidney

DCPIB:

4-(2-butyl-6, 7-dichloro-2-cyclopentyl-indan-1-on-5-yl) oxobutyric acid

HUVECs:

human umbilical vein endothelial cells

EGTA:

ethylene glycol-O,O′-bis(2-aminoethyl)-N,N,N′,N′-tetraacetic acid

HEPES:

N-(hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)

IC50 :

half-maximal concentrations

E max :

maximum effect

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Acknowledgements

This work was supported by the Research Foundation of Education Bureau of Hebei Province (BJ2018030 to XZ), the Natural Science Foundation of Hebei Province (H2015423014 to XZ), the National Natural Science Foundation of China (31401003 to XZ, 81500057 to HZ), the project funded by China Postdoctoral Science Foundation (2015M581313 to XZ), the Research Foundation of Education Bureau of Hebei Province (QN2017113 to PC, QN2016166 to WL), and the Research Foundation of Administration of Traditional Chinese Medicine of Hebei Province (2017007 to YL).

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Authors and Affiliations

  1. Department of Pharmacology, Hebei University of Chinese Medicine, No. 326 South Xinshi Road, Shijiazhuang, 050091, Hebei, China

    Yucong Xue, Xue Han, Gaohua Zhang, Yue Lin, Pingping Chen, Xiaorun Sun, Yalei Liu, Li Chu, Jianping Zhang & Xuan Zhang

  2. Department of Respiratory, The Second Hospital of Hebei Medical University, Shijiazhuang, China

    Honglin Li & Huiran Zhang

  3. Department of Medicinal Chemistry, Hebei University of Chinese Medicine, Shijiazhuang, China

    Yuanyuan Zhang & Li Chu

  4. Department of Physiology, Hebei University of Chinese Medicine, Shijiazhuang, China

    Wenya Li

  5. Department of Radiotherapy, Cangzhou Central Hospital, No. 16 West Xinhua Road, Cangzhou, China

    Mingyun Zhang

Authors
  1. Yucong Xue
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  2. Honglin Li
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Contributions

Participated in research design: X. Zhang, M. Zhang, J. Zhang, Xue, and Chu

Conducted experiments: Xue, H. Li, Y. Zhang, Han, G. Zhang, W. Li, Lin, H. Zhang, Chen, and Sun

Performed data analysis: Xue, G. Zhang, and Liu

Wrote or contributed to the writing of the manuscript: X. Zhang, M. Zhang, J. Zhang, and Xue

Corresponding authors

Correspondence to Jianping Zhang, Mingyun Zhang or Xuan Zhang.

Additional information

This article is part of the Topical Collection on Ion channels, receptors and transporters

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Figure S1

The effects of five natural flavonoids on VRAC currents in HUVEC cells. (PDF 309 kb)

(A) Representative current traces for the effects of five natural flavonoids (100 μM) on VRAC currents tested at −100 mV in HUVEC cells.

(B) The mean inhibition of VRAC current tested at −100 mV by five natural flavonoids (100 μM) in HUVEC cells. *p < 0.05 compared with the current amplitudes in the absence of drugs.

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Xue, Y., Li, H., Zhang, Y. et al. Natural and synthetic flavonoids, novel blockers of the volume-regulated anion channels, inhibit endothelial cell proliferation. Pflugers Arch - Eur J Physiol 470, 1473–1483 (2018). https://doi.org/10.1007/s00424-018-2170-8

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  • DOI: https://doi.org/10.1007/s00424-018-2170-8

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