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Evaluating lipopolysaccharide-induced oxidative stress in bovine granulosa cells

  • Reproductive Physiology and Disease
  • Published:
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Abstract

Purpose

The purpose of this study was to evaluate the capacity of bovine granulosa cells to generate reactive oxygen intermediates in response to lipopolysaccharide. We hypothesized that granulosa cells increase reactive oxygen intermediates in response to Gram-negative lipopolysaccharide in a similar manner to immune cells.

Methods

Bovine peripheral blood mononuclear cells and granulosa cells were cultured in the presence of lipopolysaccharide. Oxidative stress was evaluated using the fluorescent marker dye CellROX, and oxidative stress-related genes were measured using real-time RT-PCR.

Results

As expected, peripheral blood mononuclear cells increased oxidative stress in response to lipopolysaccharide as measured by accumulation of the fluorescent marker dye CellROX. While granulosa cells demonstrate the capacity to increase accumulation of CellROX dye in response to a positive control menadione, lipopolysaccharide had no effect on accumulation of CellROX dye. The expression of GSR, SOD1, and SOD2 were variable in peripheral blood mononuclear cells treated with lipopolysaccharide but were consistently upregulated when co-incubated with the antioxidant, N-acetyl cysteine. The expression of oxidative stress­related genes was not altered in granulosa cells, with the exception of elevated SOD1 following lipopolysaccharide exposure in the absence of antioxidant.

Conclusions

Combined, these data suggest that while reactive stress is important in pathogen killing and inflammation in immune cells, granulosa cells do not increase oxidative stress in response to lipopolysaccharide.

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Acknowledgements

The authors wish to thank Dr. John Driver for his support and valuable guidance in completing the flow cytometry analysis. Research reported in this publication was supported by the Eunice Kennedy Shriver National Institute of Child Health & Human Development of the National Institutes of Health under Award Number R01HD084316.

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Authors and Affiliations

  1. Department of Animal Sciences, University of Florida, PO Box 110910, Gainesville, FL, 32611-0910, USA

    John J. Bromfield & Sossi M. Iacovides

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  1. John J. Bromfield
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  2. Sossi M. Iacovides
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Correspondence to John J. Bromfield.

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The authors declare that they have no conflict of interest.

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Bromfield, J.J., Iacovides, S.M. Evaluating lipopolysaccharide-induced oxidative stress in bovine granulosa cells. J Assist Reprod Genet 34, 1619–1626 (2017). https://doi.org/10.1007/s10815-017-1031-2

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  • DOI: https://doi.org/10.1007/s10815-017-1031-2

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