Progesterone modulates the expression of interleukin-6 in cultured term human uterine cervical fibroblasts.

PROBLEM: The preventative value of progesterone in preterm labor has been recently recognized, especially when it is administered via vaginal suppository. This study was undertaken to evaluate the effect of progesterone on interleukin-6 (IL-6) production in human uterine cervical fibroblasts (UCFs) treated with lipopolysaccharides (LPS). METHOD OF STUDY: Human uterine cervical tissue was obtained at term, prior to the onset of labor, during the scheduled cesarean section or cesarean hysterectomy. Primary UCF cultures were established and confirmed by immunohistochemistry. IL-6 mRNA and protein expressions were examined by reverse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: Lipopolysaccharides stimulation induced a clear time- and dose-dependent increase in IL-6 mRNA and protein levels in UCFs (P < 0.05). Progesterone treatment significantly attenuated LPS-induced increases in IL-6 mRNA and protein expressions in UCFs (P < 0.05). Estrogen exposure had no effect on LPS-induced IL-6 up-regulation and did not modulate the effects of progesterone. CONCLUSION: Our preliminary results suggest that vaginal progesterone might prevent spontaneous preterm labor through a mechanism involving anti-inflammatory effects on UCFs, particularly suppression of IL-6 production.

Urotensin-II 143 G/A polymorphism is not associated with the risk of preeclampsia in Korean women.

PROBLEM: Urotensin-II (UTS-II) may be associated with preeclampsia. We therefore assessed whether the UTS-II 143 G/A polymorphism is associated with preeclampsia in Korean women. METHOD OF STUDY: This prospective case-control study enrolled 109 patients with preeclampsia and 144 healthy pregnant controls, all of whom were genotyped for the UTS-II 143 G/A polymorphism using the real-time TaqMan SNP Genotyping Assay. RESULTS: There was no significant difference in the distribution of the 143 G/A polymorphism or the frequency of the 143 A allele between women with preeclampsia and controls. In a dominant model, carriers of the A allele were not significantly more prevalent in the preeclamptic (53.2%) than in the control (60.4%) group. In addition, subgroup analysis showed no significant difference in genotype distribution or allelic frequency of the 143 G/A polymorphism between women with mild or severe preeclampsia and controls. CONCLUSION: In Korean women, the common UTS-II 143 G/A polymorphism is unlikely to have an association with the risk of preeclampsia.

Vascular endothelial growth factor gene +936 C/T polymorphism is associated with preeclampsia in Korean women.

OBJECTIVE: The purpose of this study was to evaluate potential associations between vascular endothelial growth factor (VEGF) gene polymorphisms and preeclampsia. STUDY DESIGN: One hundred ten patients with preeclampsia and 209 healthy pregnant control subjects were enrolled in the study. After peripheral blood was obtained from all women and the genomic DNA was isolated, we genotyped +936C/T polymorphisms in the 3'-untranslated region of the VEGF gene, using polymerase chain reaction and restriction fragment length polymorphism techniques. RESULTS: The distribution of genotypes of the +936C/T polymorphism was significantly different between women with preeclampsia and the control group (P < .001). Carriage of the +936T allele was significantly more frequent in preeclamptic patients than in control subjects (odds ratio, 2.06; 95% CI,1.38-3.08). Logistic regression analysis on VEGF genotype and clinical parameters such as age, educational status, body mass index, and neonatal gender showed carriage of the 936T allele to be significantly more frequent in preeclamptic patients than in control subjects (adjusted odds ratio, 2.23; 95% CI, 1.46-3.42). CONCLUSION: Carriage of the +936T allele of the VEGF gene may be associated with increased susceptibility to the development of preeclampsia and may be an independent risk factor for preeclampsia.

Effect of dehydroepiandrosterone on lipopolysaccharide-induced interleukin-6 production in DH82 cultured canine macrophage cells.

OBJECTIVE: This study examined the effect of dehydroepiandrosterone (DHEA) on lipopolysaccharide (LPS)-stimulated interleukin-6 (IL-6) production in the DH82 canine macrophage cell line. STUDY DESIGN: Cultured DH82 cells were stimulated with varying concentrations of LPS with or without DHEA for various times. Supernatant IL-6 levels were measured by enzyme-linked immunosorbent assay, and cellular cytoplasmic IkappaBalpha protein expression measured by Western blot analysis. RESULTS: LPS dose-dependently stimulated IL-6 production (p=0.016). Cells stimulated with 20 microg LPS showed a time-dependent increase of IL-6 concentration up to 10 h post-treatment (p=0.007). Co-treatment of DH82 cells with 20 microg LPS and various concentrations of DHEA for 14 h showed that up to 10 microM DHEA dose-dependently decreased the IL-6 concentration (p=0.007). Also, addition of 20 microM DHEA to DH82 cells with 20 microg LPS time-dependently decreased the IL-6 concentration for up to 14 h post-treatment (p=0.018). Stimulation of cultured DH82 cells with 20 microg LPS significantly decreased cellular cytoplasmic IkappaBalpha expression, beginning at 30 min post-treatment and persisting to at least 2 h post-treatment (p=0.012). However, co-treatment of cells with 20 microg LPS and 20 microM DHEA abrogated this effect until 2 h post-treatment. CONCLUSIONS: DHEA decreased the IL-6 concentration in the supernatant of LPS-stimulated DH82 cells by inhibiting the sequestration of IkappaBalpha, which is necessary for the activation of nuclear factor-kappa B. These findings provide new insights into the immunomodulatory effects of DHEA.

Therapeutic effect of cyclo-oxygenase inhibitors with different isoform selectivity in lipopolysaccharide-induced preterm birth in mice.

OBJECTIVE: Cyclo-oxygenase inhibitors have different selectivity for inhibitory action on the isoforms of the enzyme. Our purpose was to compare the tocolytic and maternal toxic effects of various cyclo-oxygenase inhibitors in the lipopolysaccharide-induced preterm birth in pregnant mice. STUDY DESIGN: We randomly assigned 50 ICR pregnant mice into five groups of 10 mice each and treated them intraperitoneally with either phosphate-buffered saline solution or lipopolysaccharide (50 microg) on day 15 of pregnancy. Of the four groups that received lipopolysaccharide, three groups also were treated either with nonselective cyclo-oxygenase inhibitors (indomethacin [1 mg/kg/d] or diclofenac [2 mg/kg/d]) or with the cyclo-oxygenase-2 preferential inhibitor, meloxicam (2 mg/kg/d), with a gavage tube on days 15 through 18 of pregnancy or until maternal death. The mice were killed immediately after preterm birth or on day 21 of pregnancy. Preterm birth rates and maternal side effect profiles were evaluated. RESULTS: Preterm birth occurred in 90% of mice after intraperitoneal lipopolysaccharide injection and in 20% of mice after phosphate-buffered saline solution injection. Indomethacin and meloxicam, but not diclofenac, significantly decreased the incidence of preterm birth that was induced by lipopolysaccharide (33.3% and 33.3%, respectively; P =.028). Although the overall incidence of maternal gastric and/or renal toxicities was not significantly increased in the indomethacin or meloxicam groups, a significant increase was noticed in the diclofenac group compared with the lipopolysaccharide-treated control group (P =.006). CONCLUSION: Indomethacin and meloxicam, but not diclofenac, inhibit the lipopolysaccharide-induced preterm birth in an animal model. Meloxicam appears to have no advantage over indomethacin with regard to tocolysis and maternal side effects.