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Abstract
Purpose
In mammals, α and β-defensins are antimicrobial peptides that are expressed in various epithelial and phagocytic cells. Human β-defensin-1 and -2 (hBD-1, hBD-2) have recently been shown to be expressed in various epithelial cells. Vaginal mucosa can be a target of vaginitis and the site of uropathogens' colonization that precedes urinary tract infections. Therefore, innate host defense mediators like antimicrobial peptides in the vaginal mucosa are important. Estrogen and progesterone receptors have been shown to be expressed in the vaginal squamous epithelium. Sex hormones like estrogen and progesterone may cause vaginal atrophy or susceptibility to uropathogens. So, we performed this study to investigate the expression patterns of hBD-1 and -2 mRNA in vaginal squamous epithelium (VSE) with using lipopolysaccharide (LPS), 17β-estradiol and progesterone.
Materials and Methods
Normal VSE cells that were retrieved from vaginal tissue during vaginoplasty were primarily cultured in keratinocyte growth medium and they we allowed to undergo their 3rd passage. Modulation of the expressions of hBD-1 and -2 mRNA by various stimuli (LPS 0.5 µg/ml, E2 2nM, P 1 µM) was measured by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).
Results
HBD-1 and -2 were constitutively expressed in the normal VSE cell lines, but the hBD-2 expression was not significant. A marked increase of the constitutive expression of hBD-2 mRNA was observed upon stimulation with LPS, but not upon stimulation with E2. A moderate decrease of the constitutive expression of hBD-2 mRNA upon stimulation with LPS was observed with administering progesterone.
Figures and Tables
Fig. 1
Expression of human β-defensin-1 mRNA. (A) RT-PCR analysis in the presence of the control (1), and 17β-estradiol 2nM (2), LPS 0.5 µg/ml (3), 17β-estradiol 2nM+LPS 0.5 µg/ml (4) after incubation for 12 hours in the vaginal squamous epithelial cells: the expression of hBD-1 mRNA was constitutive, but it showed no difference from (1)-(4). RT-PCR: reverse transcription polymerase chain reaction, LPS: lipopolysaccharide. (B) RT-PCR analysis in the presence of the control (1), and Progesteron 1 µM (2), LPS 0.5 µg/ml (3) and Progesteron 1 µM+LPS 0.5 µg/ml (4) after incubation for 12 hours in vaginal squamous epithelial cells: the expression of hBD-1 mRNA shown no difference from (1)-(4).
Fig. 2
Expression in human β-defensin-2 mRNA. (A) RT-PCR analysis of the presence of control (1) and 17β-estradiol 2nM (2), LPS 0.5 µg/ml (3) and 17β-estradiol 2nM+LPS 0.5 µg/ml (4) after incubation for 12 hours in the vaginal squamous epithelial cells: the constitutive expression of hBD-2 mRNA was insignificant and it showed a marked increase in (2), and there was no difference between (3) and (4). RT-PCR: reverse transcription polymerase chain reaction, LPS: lipopolysaccharide. (B) RT-PCR analysis in the presence of the control (1), and Progesterone 1 µM (2), LPS 0.5 µg/ml (3) and Progesterone 1 µM+LPS 0.5 µg/ml (4) after incubation for 12 hours in the vaginal squamous epithelial cells: the expression of hBD-2 mRNA in (4) was less than in (3).
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