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Effects of ouabain on proliferation, intracellular free calcium and c-myc mRNA expression in vascular smooth muscle cells

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Abstract

Ouabain, a newly discovered mammalian steroid hormone, has been shown to correlate directly with mean blood pressure. In the present study, the mechanism by which ouabain may act on blood pressure was investigated using primary cultures of bovine vascular smooth muscle cells. 3-[4,5-Dimethylthiazol-2-yl]-2,5-dephenyl tetrazolium bromide (MTT) assay showed a biphasic effect on cell proliferation: at low concentrations ouabain stimulated cell growth while at higher concentrations it inhibited cell growth, which is similar to angiotensin II, an octapeptide hormone. Confocal laser scanning microscopy assay also indicated a biphasic nature on intracellular free calcium, which was decreased by low concentration of ouabain and increased by higher concentration of ouabain. Simultaneously, an increase of c-myc mRNA expression by low concentration of ouabain was obtained with reverse transcription-polymerase chain reaction (RT-PCR) assay. It is important to note that the low concentrations (10−15–10−12 M) are under the normal circulating levels (2.5–8.0 × 10−11 M) of endogenous ouabain. These results suggest that ouabain-induced proliferation might be attributed, at least in part, to the decrease of intracellular free calcium and the increase of c-myc mRNA expression, and that may be directly or indirectly involved in the regulation of blood pressure.

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Abbreviations

AngII:

Angiotensin II

[Ca2+]i :

Intracellular calcium concentration

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

dNTP:

Deoxy-nucleotidetriphosphate

FBS:

Fetal bovine serum

MTT:

3-[4,5-Dimethylthiazol-2-yl]-2,5-dephenyl tetrazolium bromide

VSMCs:

Vascular smooth muscle cells

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Acknowledgments

We express our sincere appreciation for the technical assistance and patience of Mr Xuegang Luo, a doctor of life sciences at Life Science College, China Pharmaceutical University. We would also like to thank Dr. Chenghua Zhou of China Pharmaceutical University for assistance on primer cultures of VSMCs. This work was supported by the Health Department of Heilongjiang Province, China, 2005-316. The experiments performed comply with current Chinese laws.

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Authors and Affiliations

  1. Department of Physiology, China Pharmaceutical University, 24 Tongjia Xiang, Nanjing, 210009, People’s Republic of China

    Mengquan Li & Qiujuan Wang

  2. Department of Pharmacology, Mudanjiang Medical College, 3 Tongxiang Street, Mudanjiang City, 157011, People’s Republic of China

    Lixin Guan

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  1. Mengquan Li
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  2. Qiujuan Wang
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Correspondence to Qiujuan Wang.

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Communicated by I.D. Hume.

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Li, M., Wang, Q. & Guan, L. Effects of ouabain on proliferation, intracellular free calcium and c-myc mRNA expression in vascular smooth muscle cells. J Comp Physiol B 177, 589–595 (2007). https://doi.org/10.1007/s00360-007-0157-4

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  • DOI: https://doi.org/10.1007/s00360-007-0157-4

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