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Author:

Ren, Yan-Ping (Ren, Yan-Ping.) | Zhang, Ming-Juan (Zhang, Ming-Juan.) | Zhang, Ting (Zhang, Ting.) | Huang, Ruo-Wen (Huang, Ruo-Wen.)

Indexed by:

SCIE

Abstract:

Purpose: To elucidate the effect of ouabain on the regulation of proliferation and apoptosis of HUVECs and involvement of different Na+-K+-ATPase alpha-subunits and NF-kappa B. Methods: HUVECs were isolated by collagenase perfusion, and MTT assays and cell cycle analysis were performed to study proliferation. NF-kappa B expression and function were examined by immunohistochemical staining and western blotting. Na+-K+-ATPase activity was determined by measuring released ouabain inhibitable inorganic phosphate (Pi). The expression of different alpha-subunits was investigated by real RT-PCR, western blotting and cell immunofluorescence. Results: 0.3 nM ouabain treatment for 0.5 h triggered the proliferation of HUVECs, peaking at 1-2 h. At 1.8 nM for 0.5 h, ouabain induced an increase of cell proliferation for a short time, and then triggered a decrease after 1 h. Cell cycle analysis show that 37% of HUVECs were in G2/M phase of the cell cycle following incubation with 1.8 nM ouabain, compared with 18% with 0.3 nM ouabain. NF-kappa B activity was assessed by western blot analysis of I kappa B expression, which was significantly reduced with 0.3 nM ouabain treatment; there was no different between 1.8 nM ouabain treatment and untreated cells. Na+-K+-ATPase activity in HUVECs was markedly reduced after treatment with 0.3 nM and 1.8 nM ouabain. Real RTPCR and western blotting indicated that Na+-K+-ATPase alpha(1)-subunit mRNA expression levels increased after 0.3 nM ouabain treatment and decreased after 1.8 nM ouabain treatment. However, alpha(2)- and alpha(3)-subunit mRNA decreased after 0.3 nM ouabain treatment and increased after 1.8 nM ouabain treatment. Conclusion: Ouabain at different concentrations caused dual effects on proliferation and apoptosis in HUVECs.

Keyword:

NF-kappa B Ouabain alpha subunit endothelium

Author Community:

  • [ 1 ] [Ren, Yan-Ping; Zhang, Ting; Huang, Ruo-Wen] Xi An Jiao Tong Univ, Dept Geriatr Cardiovasc Dis, Affiliated Hosp 1, Coll Med, Xian 710061, Peoples R China
  • [ 2 ] [Zhang, Ming-Juan] Xi An Jiao Tong Univ, Coll Med, Affiliated Hosp 2, Dept Cardiol, Xian 710004, Peoples R China

Reprint Author's Address:

  • Xi An Jiao Tong Univ, Dept Geriatr Cardiovasc Dis, Affiliated Hosp 1, Coll Med, Xian 710061, Peoples R China.

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Source :

INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE

ISSN: 1940-5901

Year: 2014

Issue: 5

Volume: 7

Page: 1214-1222

1 . 2 7 7

JCR@2014

0 . 1 8 1

JCR@2018

ESI Discipline: BIOLOGY & BIOCHEMISTRY;

ESI HC Threshold:122

JCR Journal Grade:4

CAS Journal Grade:4

Cited Count:

WoS CC Cited Count: 10

SCOPUS Cited Count:

ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 12

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