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Updated: Jul 17, 2025
A Tissue Culture Model of Estrogen-producing Primary Bovine Granulosa Cells
Published on: September 6, 2018
β-Hydroxybutyrate affects cell physiological parameters, inflammatory markers and hormone receptor expression in
Kim Kraushaar1, Julia Hollenbach1, Niklas Schmauch1
1Institute of Anatomy, University of Veterinary Medicine Hannover Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany.
Introduction:
After calving, dairy cows are commonly affected by negative energy balance (NEB), indicated by high β-Hydroxybutyrate (BHBA) blood levels. These are associated with subfertility frequently related to uterine inflammation. Since this could compromise functionality of endometrial glands that are essential for proper embryo implantation in sheep, we investigated effects of BHBA on bovine endometrial gland cells (BEGC) in vitro.
Material And Methods:
BEGC were stimulated with different concentrations of BHBA over different periods. Cell metabolism and motility were examined by MTT-assay and Live-cell-imaging. The mRNA expression of the receptors for estrogen (ESR1, ESR2), progesterone (PR) and IFNτ (IFNAR1, IFNAR2), and the inflammatory cytokines TNFα and IL-6 was determined by RT-qPCR. Protein expression for PR and ESR1 was analyzed by semiquantitative Western Blot.
Results:
BEGC metabolism was significantly decreased after stimulation with 1.2, 1.8 and 2.4 mM BHBA over 24 and 36 h. Cell motility was significantly reduced by 1.8 and 2.4 mM BHBA already after 11 h. After 24 h stimulation, the ESR1 mRNA expression was significantly increased in BEGC stimulated with 0.6 mM BHBA. PR and TNFα mRNA expressions were increased in cells stimulated with 2.4 mM BHBA. Protein expression of ESR1 and PR was not altered.
Discussion:
Treatment with BHBA leads to restriction of BEGC metabolism and motility, and increased expression of TNFα, ESR1 and PR in vitro. This could explain how increased BHBA blood levels might compromise functionality of uterine glands in vivo and thus could contribute to compromised reproductive success of cows suffering from NEB.
