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The contrasting effect of estrogen on mRNA expression of VEGF in bovine retinal vascular endothelial cells under different oxygen conditions

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Abstract

Objective

To investigate the influence of Estradial (E2) at physiological concentration on mRNA expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α(HIF-α1) in bovine retinal vascular endothelial cells (BRECs) under different oxygen conditions.

Methods

The mRNA expression of VEGF, HIF-1α in BRECs was studied by quantitative reverse-transcription PCR (qRT-PCR), and protein levels were assayed by Western Blot. Different concentrations of E2 (10−12, 10−10, 10−8 mol/l) and estrogen receptor antagonist Tamoxifen (10−7 mol/l) were added into the cell culture medium of different groups, while the phosphate-buffered saline (PBS) was added in the control group instead. Culture conditions were set to be under normoxia and hypoxia. The results of each group were detected after 8 hours and 24 hours.

Results

(1) Under hypoxia, gene expression of VEGF, HIF-1α in BRECs increased more than that of the control group (P < 0.05). There is evident positive correlation between the mRNA and protein levels of VEGF and HIF-1α (r = 0.82). (2) Treated with 10–8 mol/l E2, the expression of VEGF mRNA was increased in a time–dependent manner (P < 0.05). Treated with the indicated to concentrations of E2 (10− 12 ∼ − 8 mol/l) under normoxia for twenty-four hours, the mRNA expression of VEGF in BRECs was increased in a dose–dependent manner (P < 0.05), whereas E2 did not influence the mRNA expression of HIF-1α (P > 0.05). (3) Under hypoxia , E2 reduced the mRNA and protein levels of HIF-1α and VEGF in a concentration-dependent manner (P < 0.05), and the decrease developed in a time-dependent manner (P < 0.05). (4) An overdose of tamoxifen (10−7 mol/l) reversed the effect of E2 (10−8 mol/l) (P < 0.05).

Conclusions

E2 increases the gene expression of VEGF in BRECs under normoxia, whereas E2 reduces the gene expression of VEGF in BRECs through HIF-1α under hypoxic conditions in a dose-dependent and time-dependent manner. The contrasting effect of E2 on mRNA expression of VEGF may play a prophylactic role in retinopathy of prematurity (ROP).

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Acknowledgments

The present study was conducted thanks to the staff of the central laboratory of Shandong Provincial Hospital for cell culture assistance.

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Authors and Affiliations

  1. Shandong Shierming Eye Hospital, 48 Jinan Ying Xiong Shan Road, Jinan, 250002, People’s Republic of China

    Xiaohua Ma, Hongsheng Bi & Xiaofeng Xie

  2. Eye Institute of Shandong University of TCM, 48 Jinan Ying Xiong Shan Road, Jinan, 250002, People’s Republic of China

    Yi Qu

  3. Department of Ophthalmology, Qilu Hospital of Shan dong University, Jinan, 250012, People’s Republic of China

    Jinghai Li

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  1. Xiaohua Ma
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  2. Hongsheng Bi
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  3. Yi Qu
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  4. Xiaofeng Xie
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Corresponding author

Correspondence to Hongsheng Bi.

Additional information

There is no financial relationship with the origination that sponsored our research. We certify that we have full control of all primary data and agree to allow Graefes Archive for Clinical and Experimental Ophthalmology to review our data upon request.

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Ma, X., Bi, H., Qu, Y. et al. The contrasting effect of estrogen on mRNA expression of VEGF in bovine retinal vascular endothelial cells under different oxygen conditions. Graefes Arch Clin Exp Ophthalmol 249, 871–877 (2011). https://doi.org/10.1007/s00417-010-1594-4

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  • DOI: https://doi.org/10.1007/s00417-010-1594-4

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