Increased cystic fibrosis transmembrane conductance regulators expression and decreased epithelial sodium channel alpha subunits expression in early abortion: Findings from a mouse model and clinical cases of abortion

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Abstract

The status of the maternal endometrium is vital in regulating humoral homeostasis and for ensuring embryo implantation. Cystic fibrosis transmembrane conductance regulators (CFTR) and epithelial sodium channel alpha subunits (ENaC-α) play an important role in female reproduction by maintaining humoral and cell homeostasis. However, it is not clear whether the expression levels of CFTR and ENaC-α in the decidual component during early pregnancy are related with early miscarriage. CBA6DBA/2 mouse mating has been widely accepted as a classical model of early miscarriage. The abortion rate associated with this mating was 33.33% in our study. The decidua of abortion-prone CBA female mice (DBA/2 mated) had higher CFTR mRNA and protein expression and lower ENaC-α mRNA and protein expression, compared to normal pregnant CBA mice (BLAB/C mated). Furthermore, increased CFTR expression and decreased ENaC-α expression were observed in the uterine tissue from women with early miscarriage, as compared to those with successful pregnancy. In conclusion, increased CFTR expression and decreased ENaC-α expression in the decidua of early abortion may relate with failure of early pregnancy. © 2014 Zhou et al.

Figures

  • Table 1. Sequences of the primers used for qRT-PCR analysis.
  • Figure 1. Findings for the CBA6DBA/2 abortion model. (A) In the CBA6DBA/2 group (n = 11), 30 abortion-prone embryos were obtained, while in the control CBA6BALB/C group (n = 10), six abortion-prone embryos were obtained. (B) Implantation rates in the experimental group (8.1862.14) and control group (7.9061.79) (P.0.05). (C) Vaginal bleeding was observed on days 7–10 of pregnancy. (D) Representative image of abortion-prone and normal embryos: the arrows indicate the abortion-prone embryos with dysplasia appearance, in which fetuses and placentas can be hardly identified from each other. (E, F) Cross-sections of uterine tissue at the implantation site showed typical ischemia, hemorrhage, and necrosis in abortion-prone embryos (E), but not in healthy embryos (F) (scale bar: 300 mm). Green arrows indicate hemorrhage, and blue arrows indicate necrosis. doi:10.1371/journal.pone.0099521.g001
  • Figure 2. CFTR and ENaC-a mRNA expression in decidua tissue. CFTR mRNA expression increased and ENaC-a mRNA expression decreased in the decidua of CBA6DBA/2 mice, and the difference in expression was statistically significant compared to the CBA6BALB/C mice (A). Similarly, higher CFTR mRNA expression and lower ENaC-a mRNA expression were detected in the decidua of women who underwent abortion (B). **P,0.01. doi:10.1371/journal.pone.0099521.g002
  • Figure 3. Immunohistochemical analysis of CFTR expression in mouse and human tissue samples. CFTR protein is indicated by arrows in the image. Representative images (4006 for the insets) are shown. CFTR was detected as brown signals located mainly to the cell membrane. The expression of CFTR protein was stronger in the decidua of CBA6DBA/2 mice than CBA6BALB/C mice; moreover, it was stronger in women who had a miscarriage than normal pregnant women. Rat lung tissue was used as the positive control. Negative controls were obtained by replacing the first antibody with PBS. (scale bar: 50 mm). doi:10.1371/journal.pone.0099521.g003
  • Figure 4. Immunohistochemical analysis of ENaC-a staining in mouse and human tissue samples. ENaC-a is indicated by arrows in the image. Representative images (4006 for the insets) are shown. ENaC-a was detected as brown signals located mainly to the cell membrane. The expression of ENaC-a protein was weaker in the decidua of the CBA6DBA/2 mice compared to the CBA6BALB/C mice; moreover, it was weaker in the women who underwent miscarriage than normal pregnant women. Rat lung tissue was used as the positive control. Negative controls were obtained by replacing the first antibody with PBS (scale bar: 50 mm). doi:10.1371/journal.pone.0099521.g004
  • Figure 5. CFTR and ENaC-a protein detection by western blot analysis. The expression of CFTR protein was stronger and the expression of ENaC-a protein was weaker in the decidua of CBA6DBA/2 mice compared to CBA6BALB/C mice (A). Moreover, the expression of CFTR protein was stronger and the expression of ENaC-a protein was weaker in the decidua of women who underwent a miscarriage than normal pregnant women (B). b-tubulin was used as the internal control (approximately 43 kD). Representative images are shown. Densitometry analysis data for CFTR and ENaC-a expression are normalized to the control (100%) for comparison. *P,0.05. doi:10.1371/journal.pone.0099521.g005

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Zhou, M., Fu, J., Huang, W., Shen, L., Xiao, L., Song, Y., & Liu, Y. (2014). Increased cystic fibrosis transmembrane conductance regulators expression and decreased epithelial sodium channel alpha subunits expression in early abortion: Findings from a mouse model and clinical cases of abortion. PLoS ONE, 9(6). https://doi.org/10.1371/journal.pone.0099521

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