Pilot Study on the Effect of Patient Condition and Clinical Parameters on Hypoxia-Induced Factor Expression: HIF1A, EPAS1 and HIF3A in Human Colostrum Cells

Abstract

Hypoxia-inducible factor 1 (HIF-1) may play a role in mammary gland development, milk production and secretion in mammals. Due to the limited number of scientific reports on the expression of HIF genes in colostrum cells, it was decided to examine the expression of HIF1A, HIF3A and EPAS1 in the these cells, collected from 35 patients who voluntarily agreed to provide their biological material for research, were informed about the purpose of the study and signed a consent to participate in it. The expression of HIF genes was assessed using qPCR. Additionally, the influence of clinical parameters (method of delivery, occurrence of stillbirths in previous pregnancies, BMI level before pregnancy and at the moment of delivery, presence of hypertension during pregnancy, presence of Escherichia coli in vaginal culture, iron supplement and heparin intake during pregnancy) on the gene expression was assessed, revealing statistically significant correlations. The expression of HIF1A was 3.5-fold higher in the case of patients with the presence of E. coli in vaginal culture (p = 0.041) and 2.5 times higher (p = 0.031) in samples from women who used heparin during pregnancy. Approximately 1.7-fold higher expression of the EPAS1 was observed in women who did not supplement iron during pregnancy (p = 0.046). To our knowledge, these are the first studies showing the relationship between HIF expression in cells from breast milk and the method of delivery and health condition of women giving birth. The assessment of HIF expression requires deeper examination in a larger study group, and the results of further studies will allow to determine whether HIF can become biomarkers in pregnancy pathology states.

Keywords: EPAS1; HIF1A; HIF3A; colostrum; gene expression; hpoxia-inducible factor; human milk.

Figure 1

(A) Average expression of the HIF3 gene (RQ ± SE) in the cellular fraction of breast milk depending on the method of delivery, (B) Average expression of the EPAS1 gene (RQ ± SE) in the cellular fraction of breast milk depending on the occurrence of stillbirths in previous pregnancies, (C) Average expression of the HIF3A gene (RQ ± SE) in the cellular fraction of breast milk depending on the BMI before pregnancy, (D) Average expression of the HIF3A gene (RQ ± SE) in the cellular fraction of breast milk depending on the BMI at the moment of delivery, (E) Average expression of the EPAS1 gene (RQ ± SE) in the cellular fraction of breast milk depending on the occurrence of hypertension during the pregnancy, (F) Average expression of the HIF1A gene (RQ ± SE) in the cellular fraction of breast milk depending on the presence of Escherichia coli in vaginal culture, (G) Average expression of the HIF3 gene (RQ ± SE) in the cellular fraction of breast milk depending on the presence of comorbidities, (H) Average expression of the EPAS1 gene (RQ ± SE) in the cellular fraction of breast milk depending on the iron supplementation by the patient, (I) Average expression of the HIF1A gene (RQ ± SE) in the cellular fraction of breast milk depending on the heparin treatment, * p < 0.05, ** p < 0.01, *** p ≤ 0.001 as determined by a Mann–Whitney U test.

Figure 2

(A) Scatter graph of the body weight before pregnancy and the expression (RQ) of the HIF3A gene in the cellular fraction of breast milk (r = −0.509 p < 0.05); (B) Scatter graph of the BMI before pregnancy and the expression (RQ) of the HIF3A gene in the cellular fraction of breast milk (r = −0.509 p < 0.05); (C) Scatter graph of the current BMI and the expression (RQ) of the HIF3A gene in the cellular fraction of breast milk (r = −0.509 p < 0.05); (D) Scatter graph of the PLT (child) and the expression (RQ) of the HIF3A gene in the cellular fraction of breast milk (r = −0.509 p < 0.05); (E) Scatter graph of the glucose (child) and the expression (RQ) of the HIF1A gene in the cellular fraction of breast milk (r = −0.509 p < 0.05) Spearman’s Rank Correlation.