Different regulation of oestrogen receptors alpha and beta in the human cervix at term pregnancy.

During pregnancy, a cervical connective tissue remodelling takes place, clinically recognized as softening, effacement and dilatation. The role of oestrogens and their receptors (ER) in this process is not clear. ERalpha is a ligand-activated transcription factor involved in many physiological processes. The identification of a second oestrogen receptor, ERbeta, has led to a re-evaluation of oestrogen signalling and physiology. The aim of this study was to monitor the expression of the two ERs in the cervix from women at term pregnancy (TP) and after parturition (PP) compared with that of non-pregnant (NP). A solution hybridization assay showed that the level of ERalpha mRNA was significantly decreased in the PP group, when compared with the NP and TP groups. In contrast the ERbeta mRNA level was increased in the TP group compared with the NP and PP groups. These results were supported by reverse transcription-polymerase chain reaction (RT-PCR). Similar results were observed for the protein with immunohistochemistry. Intense ERbeta immunostaining was observed in neutrophils and the endothelial cells of blood vessels. In conclusion, this study supports a concept according to which oestrogen might be involved in the final remodelling of the cervix via the modulating effects of the two ERs. Furthermore, oestrogen may mediate some effects on cervical ripening via ERbeta present in the invading neutrophils. Further studies are needed to elucidate this finding.

The expression of glutaredoxin is increased in the human cervix in term pregnancy and immediately post-partum, particularly after prostaglandin-induced delivery.

Glutaredoxins are glutathione disulphide oxidoreductases catalysing disulphide reductions via a redox active disulphide. We have examined the presence of glutaredoxin in the human cervix, and its differential expression during cervical remodelling in term pregnancy and immediately post-partum as compared to the non-pregnant state. Cervical biopsies were obtained from 24 term-pregnant and 24 post-partal women, of which 10 were taken after spontaneous delivery, 10 after prostaglandin-induced delivery and four after mifepristone-induced delivery, all obtained within 15 min after delivery. Six non-pregnant women served as controls. The tissues were analysed for the glutaredoxin mRNA levels using a solution hybridization method. Glutaredoxin mRNA was expressed in the human cervix, the level increased > or =2-fold at term pregnancy and immediately post-partum. The level of cervical glutaredoxin mRNA from prostaglandin E(2)-treated women was 3-fold higher than after spontaneous ripening and delivery. Localization of glutaredoxin was visualized with immunohistochemistry in cervices from two post-partal women, and was compared to that of thioredoxin. We conclude that glutaredoxin may be involved in the regulation of cervical ripening in humans, particularly in the inflammatory reaction seen during this process. Glutaredoxin mRNA levels are up-regulated after prostaglandin treatment, which is effective and the most commonly used substance for cervical priming and induction of labour.

Neurochemical and cellular markers in human cervix of late pregnant, postpartal and non-pregnant women.

BACKGROUND: The aim was to evaluate the peptidergic innervation and the dendritic cell content in the cervix uteri. METHODS: Cervical biopsies were obtained from late pregnant (n=5), postpartal (n=5) and non-pregnant (n=5) women. The samples were prepared for immunohistochemistry using antibodies to protein S-100 (S-100), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), human peptide histidine isoleucine amide (PHM 27), neuropeptide tyrosine (NPY), and human histocompatibility complex class II subregion DR (HLA-DR). RESULTS: Nerve fibers positive for protein S-100, and dendritic cells positive for S-100 and HLA-DR were abundant in the cervix, especially at late pregnancy. CGRP, VIP, PHM-27 and NPY positive nerve fibers were present in non-pregnant, short nerve fibers and scattered immunoreactivity at term, and further scattered immunoreactivity after parturition. NPY positive nerve fibers were decreased at term, and after parturition a scattered immunoreactivity was observed. CONCLUSIONS: The abundant protein S-100 positive nerve fibers implies an impact of myelinated nerves in the cervix uteri during pregnancy. The abundant dendritic cells, positive for HLA-DR and S-100, especially at term, indicates a general activation of the immune system until late pregnancy and parturition. The changed occurrence and distribution of immunoreactivity for CGRP, VIP and PHM-27 suggest a release of these neuropeptides until term. The changes in NPY immunoreactivity indicate a release of NPY around parturition.

Cervical ripening after treatment with prostaglandin E2 or antiprogestin (RU486). Possible mechanisms in relation to gonadal steroids.

OBJECTIVE: To compare the mechanisms for cervical ripening after treatment with prostaglandin E2 or antiprogestin (RU486) to spontaneous cervical ripening, with focus on gonadal steroid receptors. STUDY DESIGN: Cervical biopsies were obtained from postpartal women after treatment with prostaglandin E2 (n=10), or antiprogestin (n=5). Postpartal women after spontaneous cervical ripening (n=10) served as controls. Levels of estrogen and progesterone receptors, their mRNAs, insulin-like growth factor I mRNA and serum estradiol and progesterone were quantitated. The collagen concentration and solubility by pepsin were determined. Statistical tests used were Kruskal-Wallis and Mann-Whitney U test. RESULTS: After prostaglandin E2 treatment the collagen concentration was higher (P<0.05) as compared to spontaneous ripening. After antiprogestin treatment the estrogen receptor concentration was higher (P<0.05) in comparison to spontaneous ripening. CONCLUSION: The elevated estrogen receptor concentration after antiprogestin treatment, in contrast to spontaneous ripening, and prostaglandin E2 treatment, indicates a that a receptor-mediated progesterone withdrawal does not explain the events behind spontaneous cervical ripening at parturition.

Protein gene product 9.5-immunoreactive nerve fibers and cells in human cervix of late pregnant, postpartal and non-pregnant women.

BACKGROUND: The aim of this study was to examine the occurrence and distribution of the general neuronal marker protein gene product (PGP) 9.5 in the cervix uteri. METHODS: Cervical biopsies were obtained from late pregnant (n=5), postpartal (n=5) and non-pregnant (n=5) women. The samples were prepared for immunohistochemistry using antibodies to PGP 9.5. RESULTS: Nerve fibers were found consistently in all biopsies. There were differences in the occurrence and distribution of PGP 9.5 immunoreactive nerve fibers and cells between the three groups. Immunoreactive nerve fibers were observed at a moderate to abundant frequency in the stroma and around arterial vessel walls, in all groups. Immunoreactive nerve fibers were also observed at high frequency within and around glandular epithelium in the late pregnant and postpartal groups. PGP 9.5 immunoreactive cells were seen occasionally in the stroma of the non-pregnant group, but at a high frequency in the stroma, around arterial blood vessel walls, around and within the glandular epithelium in the late pregnant and postpartal groups. The total frequency of immunoreactive nerve fibers and cells was the highest in the late pregnant group, slightly lower in the postpartal group, and the lowest in the non-pregnant group. CONCLUSIONS: These findings show that changes in the general innervation take place during human cervical ripening until late pregnancy and parturition.

Differential expressions of mRNA for proteoglycans, collagens and transforming growth factor-beta in the human cervix during pregnancy and involution.

During pregnancy and involution, an extensive remodelling of the human cervical connective tissue occurs. This cervical ripening is one of the most pronounced physiological remodelling processes known in human connective tissue. To investigate how the remodelling is accomplished, the levels of mRNA for collagen I and III, versican and three small proteoglycans, biglycan, decorin and fibromodulin, were evaluated using Northern blots at different stages of cervical ripening. In the corresponding biopsies the concentration of collagen and of small and large proteoglycans were determined. The role of transforming growth factor-beta (TGF-beta) as a mediator of the remodelling process was also investigated. The concentration of collagen decreased and 1 week before partus, 50% of the nonpregnant level was attained. No further decrease was noted after partus. The mRNA for collagen I and III did, however, not decrease in the term pregnant cervix 1 week before partus. Only 20-30% decrease during the final ripening just before partus was recorded. Neither did the mRNA levels of the small proteoglycans change significantly during the ripening, despite an almost 50% decrease in the concentration of the small proteoglycans. The message for versican was, however, 5-fold increased at partus and then gradually returned to nonpregnant levels within 4 days after delivery. These changes corresponded to similar changes in the concentration of the large proteoglycan. Thus, the remodelling of the cervical connective tissue is achieved by two different mechanisms, on one hand an increased turnover of collagen and the small proteoglycans, on the other a changed transcription followed by an increased production of versican. During the involution 2- to 3-fold increases in the messages for collagen I and III, and the small proteoglycans, biglycan and decorin, corresponded to increases in the concentration of the small proteoglycans and non-extractable collagen. The message for TGF-beta was increased 2-fold immediately after delivery compared with the term pregnant state. Thus, TGF-beta may be of importance for the reconstruction of the cervix, which starts immediately after partus.

Potential roles for gonadal steroids and insulin-like growth factor I during final cervical ripening.

OBJECTIVE: To determine whether gonadal steroids and insulin-like growth factor I influence the final cervical remodeling during parturition. METHODS: Cervical biopsies were obtained transvaginally before labor (n = 10) and after spontaneous cervical ripening and vaginal delivery (n = 20). Levels of estrogen and progesterone receptors, their messenger RNAs, insulin-like growth factor I messenger RNA, and serum estradiol and progesterone were measured. Collagen and proteoglycan concentrations and compositions were measured to estimate the degree of cervical ripeness. RESULTS: The concentrations of estrogen and progesterone receptors decreased in comparison with the clinically unripe cervix before labor. The median estrogen receptor concentration (range) decreased from 10 (2-18) to 4.5 (2-14) fmol/mg protein (P < .01), and the progesterone receptor concentration from 105.5 (32-153) to 74 (30-115) fmol/mg protein (P < .05), whereas their messenger RNA levels were unchanged. The insulin-like growth factor I messenger RNA concentration declined from 16.1 (8.4-20.4) at term to 8.9 (1.5-18.5) amol/microgram DNA after parturition (P < .01). The collagen solubility by pepsin increased, but not significantly, and the collagen concentration was unchanged. The concentration of small proteoglycans, mainly decorin, decreased from 1.59 (1.20-1.97) to 0.84 (0.24-1.41) micrograms/mg wet weight (P < .001), and the concentration of versican increased, but not significantly (P = .07). CONCLUSION: Concentrations of estrogen and progesterone receptors and insulin-like growth factor I messenger RNA were decreased significantly after spontaneous cervical ripening in comparison to levels before labor. These changes coincided with a tendency toward increased collagen solubility and a decline in concentration of small proteoglycans, which probably alters collagen organization, thus allowing for cervical softening and dilation. These observations suggest that gonadal steroids influence the final cervical remodeling during parturition, an influence perhaps mediated by insulin-like growth factor I.

The expression of thioredoxin mRNA is increased in the human cervix during pregnancy.

The classic function for thioredoxin is to act as a hydrogen donor for the enzyme ribonucleotide reductase, which is essential for DNA synthesis. In addition, thioredoxin participates in the regulation of different metabolic processes via thiol redox control. These kind of processes involve changes in the activity of different enzymes, receptors or transcription factors via dithiol/disulphide interchange reactions. Thioredoxin is present in the human decidua and trophoblasts. This study was performed to investigate whether thioredoxin mRNA is present in the human cervix, and differently expressed during pregnancy as compared with the non-pregnant state. Cervical biopsies and serum samples were obtained from 28 late pregnant, 41 post-partum and 15 non-pregnant menstruating women. The tissues were analysed for thioredoxin mRNA content using a solution hybridization technique. The thioredoxin mRNA level increased 3-fold at late pregnancy in comparison with the non-pregnant state. No further increase was seen immediately after parturition, either after spontaneous delivery or after pharmacological induction. There was a positive correlation between the cervical thioredoxin mRNA level and the serum oestradiol concentration in the non-pregnant group. We suggest that thioredoxin mRNA in the human cervix is regulated, at least partly, by oestradiol.

Cervical ripening in humans: potential roles of estrogen, progesterone, and insulin-like growth factor-I.

OBJECTIVE: During pregnancy in humans a gradual connective tissue remodeling takes place in the cervix. The aim of this study was to examine a possible relationship between the action of gonadal steroids and growth factors and the biochemically identifiable changes in connective tissues during cervical ripening. STUDY DESIGN: Cervical biopsy specimens and serum samples were taken from 20 term pregnant and 20 nonpregnant menstruating women. Estrogen receptors and progesterone receptors were measured with enzyme immunoassays. The messenger ribonucleic acid levels for estrogen receptors, progesterone receptors, and insulin-like growth factor-I were determined by solution hybridization with human complementary deoxyribonucleic acid probes. The concentration of collagen and its solubility by pepsin digestion were measured. Statistical evaluations were done with the Student t test. RESULTS: In term pregnancy the estrogen receptor level decreased to 14% and the progesterone receptor level to 24% of nonpregnant levels (p <0.001 and p <0.01). The insulin-like growth factor-I messenger ribonucleic acid level increased 400% (p <0.01), whereas the messenger ribonucleic acid levels for estrogen receptors and progesterone receptors were unchanged. The changes coincided with a twofold decrease in collagen concentration (hydroxyproline) and a twofold increase in collagen solubility. CONCLUSION: Estrogen receptors and progesterone receptors are present in human cervix. A significant down-regulation of estrogen receptors and progesterone receptors and a fourfold increase in the insulin-like growth factor-I messenger ribonucleic acid level were registered in term pregnant cervix. These findings coincided with the remodeling of the cervical connective tissue.