RESUMEN
Objective: To explore the relationship of 11β-hydroxysteroid dehydrogenases (11β-HSDs) expression in the ovarian granulosa cells and cortisol regeneration in the ovary of polycystic ovary syndrome (PCOS). Methods: In accordance with the revised Rotterdam inclusion/exclusion criteria (2003), 24 patients in the PCOS group were included, meanwhile 21 patients in the control group were included. Follicular fluid and luteinized granulosa cells were collected from patients underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in the Center for Reproductive Medicine of Renji Hospital. Enzyme linked immunosorbent assay (ELISA) was used to detect the cortisol concentration and the cortisone levels of follicular fluid. Real-time quantitative PCR (RT-qPCR) was used to measure 11β-HSDs abundance in the luteinized granulosa cells. The expression of 11β-HSD1 and 11β-HSD2 in the luteinized granulosa cells were verified by immunofluorescence. The correlation of local cortisol level in the ovary with 11β-HSDs expression in the luteinized granulosa cells was further analyzed. Results: Compared with the control group, the cortisol level in the follicular fluid was elevated in the PCOS group [(477.3±35.3) nmol/L vs (292.0±36.4) nmol/L, P=0.014], while no change was observed in the cortisone levels in two groups. Both 11β-HSD1 and 11β-HSD2 were detected in the luteinized granulosa cells. Compared with the control group, the 11β-HSD1 mRNA abundance (P=0.033) and reductase activity (P=0.006) were increased significantly in the PCOS group, and there was no significant difference in the 11β-HSD2 expression and oxidase activity between two groups. Consistently, Pearson correlation analysis showed that the 11β-HSD1 but not 11β-HSD2 mRNA levels in the luteinized granulosa cells was correlated with the cortisol level in the follicular fluid (r2=0.347 6, P=0.001). Conclusion: Elevation of local cortisol in the ovary of PCOS patients may be related to the increased expression of 11β-HSD1 and reductase activity in the ovarian granulosa cells.
RESUMEN
Objective To study the changes and significance of 11β-HSD1 and 11β-HSD2 gene and protein expression in pla-centa of patients with gestational diabetes mellitus .Methods Thirty pregnant women with gestational diabetes mellitus (GDM ) and normal glucose tolerance (NGT ) were selected .Chemiluminescence was used to determine the serum cortisol ,neonatal cord blood cortisol and fasting insulin ect .Immunohistochemistry was used to detect the expression location of 11β-HSD1 and 11β-HSD2 in the placenta .The differential expression of 11β-HSD1 and 11β-HSD2 genes and proteins were detected by real-time PCR and Western blot .Results Compared with NGT group ,fasting insulin ,HOMA-IR ,insulin secretion index and maternal serum cortisol level were significantly increased in GDM group [(19 .95 ± 1 .05) mU/L vs .(12 .93 ± 1 .50) mU/L ,4 .54 ± 0 .67 vs .2 .87 ± 0 .43 , 80 .55 ± 6 .18 vs .53 .15 ± 5 .58 ,(1110 .00 ± 40 .91) nmol/L vs .(934 .1 ± 45 .92) nmol/L ,P<0 .05)] ,but there were no significant differences were found in fasting blood glucose and cord blood cortisol (P>0 .05) .The distribution of 11β-HSD1 in the placenta was distributed in the outer layer of villous syncytiotrophoblast ,villous interstitial and dry villi .The expression of 11β-HSD2 was con-centrated in the outer layer of villous syncytiotrophoblast .Real time-PCR and Western blot results showed that the expression of 11β-HSD1 mRNA and protein in placenta of GDM group was significantly lower than that of NGT group (0 .56 ± 0 .09 vs .1 .36 ± 0 .36 ,0 .27 ± 0 .07 vs .1 .00 ± 0 .01 ,P< 0 .05 ) ,11β-HSD2 mRNA was significantly higher than NGT group (5 .17 ± 1 .02 and 1 .21 ± 0 .34 respectively ,P<0 .05) ,but no significant difference was found in protein level (P>0 .05) .Conclusion The differenti-al expression of 11β-HSD1 and 11β-HSD2 in placenta of gestational diabetes mellitus can avoid the maternal poor pregnancy envi-ronment ,but will cause long-term harm to the fetus .