Expression Of Aromatase Cytochrome P450 Research Articles

Genital stones: Radiological, histopathological, ultrastructural, and molecular analysis in rooster

Epididymal lithiasis, characterized by the formation of stones in the epididymis, has been associated with a decline in fertility in roosters. This study aimed to investigate the reproductive performance, ultrastructural characteristics, and expression of aromatase cytochrome P450 (CYP19) and aquaporin 9 (AQP9) in aged broiler breeder roosters affected by epididymal lithiasis. X-ray analysis confirmed the presence of genital stones in both the epididymis and testicular tissue regions. While there was a significant decrease in sperm concentration in the affected roosters compared to non-affected roosters, no significant differences were observed in total and progressive sperm motility between the two groups. Furthermore, the affected roosters exhibited significant abnormalities in semen parameters, except for sperm concentration and morphology. Transmission electron microscopy (TEM) revealed the depletion and deciliation of ciliated cells in the distal efferent ductules of the epididymis in affected roosters. Additionally, the expression of CYP19 and AQP9 was found to be increased in the epididymal region of affected roosters. Notably, we report the presence of testicular stones for the first time in this study, in addition to epididymal stones. Considering the male reproductive tract lesions observed, we propose the term “genital stones” to describe these conditions. Moreover, our findings suggest that the overexpression of AQP9, which is associated with a high copy number of the CYP19 gene in the epididymal region of affected aged roosters, may contribute to the formation of genital stones by promoting increased reabsorption of fluids in the epididymis. The condensation of epididymal duct contents and reduction in the population of ciliated cells further impairs semen movement and can lead to the blockage of extra-testicular ducts, resulting in the low fertility syndrome observed in aged roosters.

Differences in expression of genes related to steroidgenesis in abdominal subcutaneous adipose tissue of pregnant women with and without PCOS; a case control study

BackgroundIt was reported that steroid-related gene expressions in the adipose tissue (AT) of women differ between women affected with polycystic ovary syndrome (PCOS) and non-PCOS. Although association between PCOS in mother and offspring’s health is a crucial issue, there are few studies focusing on AT of pregnant women suffering from PCOS. Our objectives were to determine the differences between mRNA expression levels of key steroid-converting enzymes in abdominal subcutaneous AT of pregnant women afflicted with PCOS and non-PCOS.MethodsTwelve pregnant women with PCOS (case) and thirty six non-PCOS pregnant women (control) (1:3 ratio; age- and BMI-matched) undergoing cesarean section were enrolled for the present study. Expressions of fifteen genes related to steriodogenesis in abdominal subcutaneous AT were investigated using quantitative real-time PCR.ResultsNo significant differences were detected with respect to age, BMI (prior pregnancy and at delivery day), gestational period and parity among pregnant women with PCOS and non-PCOS. Most of the sex steroid-converting genes except 17β-Hydroxysteroid dehydrogenases2 (17BHSD2), were highly expressed on the day of delivery in subcutaneous AT. Women with PCOS showed significantly higher mRNA levels of steroidgenic acute regulator (STAR; P < 0.001), cytochrome P450 monooxygenase (CYP11A1; P < 0.05), 17α-hydroxylase (CYP17A1; P < 0.05), and 11β-Hydroxysteroid dehydrogenase (11BHSD1 and 11BHSD2; P < 0.05). The expression of steroid 21-hydroxylase (CYP21) in non-PCOS was fourfold higher than those of women with PCOS (P < 0.001). There were no significant differences between relative expression of aromatase cytochrome P450 (CYP19A1), 3β-hydroxysteroid dehydrogenase (3BHSD1 and 3BHSD2), and 17BHSD family (1, 3, 5, 7, and 12) between the two groups.ConclusionThe expression levels of genes related to sex steroids metabolism were similar to age-matched and BMI- matched pregnant non-PCOS and pregnant women with PCOS at delivery day. However, the alterations in gene expressions involved in glucocorticoids and mineralocorticoids metabolism were shown. It is necessary to point out that further studies regarding functional activity are required. More attention should be given to AT of pregnant women with PCOS that was previously ignored.

Leiomyomatosis peritonealis disseminata with aromatase cytochrome P450 expression in a postmenopausal woman: a case study with literature review

Leiomyomatosis peritonealis disseminata with aromatase cytochrome P450 expression in a postmenopausal woman: a case study with literature review

Immunoexpression of aromatase cytochrome P450 and 17β-hydroxysteroid dehydrogenase in women's ovaries after menopause.

BackgroundMenopause results in a lack of regular menstrual cycles, leading to the reduction of estrogen production. On the other hand, ovarian androgen synthesis is still present at reduced levels and requires expression of several steroidogenic enzymes.MethodsThis study was performed on 104 postmenopausal women hospitalized due to uterine leiomyomas, endometriosis, and/or a prolapsed uterus. Patients were divided into three groups depending on the time from menopause. Group A patients experienced menopause 1–5 years before enrollment in the study (42 women). Group B included women who had their last menstruation 5–10 years before the study (40 women). Group C consisted of 22 women who were more than 10 years past menopause. Hysterectomy or removal of the uterine corpus with adnexa was performed during laparotomy. We evaluated the expression of aromatase cytochrome P450 (CYP 19) and 17β-hydroxysteroid dehydrogenase (17β HSD) by employing immunohistochemistry.ResultsActivity of 17β-HSD and CYP19 was demonstrated in the cytoplasm of stromal cells of postmenopausal ovaries, epithelium cells coating the ovaries, vascular endothelial cells, and epithelial inclusion cysts. However, overall expression of both 17β-HSD and CYP 19 decreased with time after menopause.ConclusionDemonstration of the activity of the key enzymes of ovarian steroidogenesis, CYP 19 and 17β-HSD, confirms steroidogenic activity in the ovaries of postmenopausal women. Nevertheless, ovarian steroidogenic activity decreases with time, and its significant decrease occurs 10 years after menopause.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) increases the expression of cyclooxygenase-2 and aromatase cytochrome P450 in human endometrium

To investigate the role of 2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD) on eutopic endometrium by stromal cell culture. Prospective experimental study. Human endometrial tissues obtained by hysterectomy specimens from 16 women with estrogen dependent diseases (endometriosis (n = 4), adenomyosis (n = 5), uterine fibroids (n = 7)) and 5 women who had undergone hysterectomy for carcinoma in situ of uterine cervix. Endometrial stromal cells were cultured under the following conditions: DMEM/F12 (10% FBS, 1 nM E2 and 100 nM P4). TCDD (10 nmol/l) and TNF-a (0.1ng/ml) were added according to experimental purposes. The protein expression of aromatase cytochrome P450 and COX-2 was examined by Western blot. In control group, the COX-2 protein in stromal cells cultured with TCDD was significantly increased (control; 0.63 ± 0.19 vs. TCDD; 0.94 ± 0.26, P<0.05) And the aromatase P450 protein in stromal cells cultured with TCDD and TCDD plus TNF-a was significantly increased. (control; 0.60 ± 0.26 vs. TCDD; 0.90 ± 0.36, TCDD plus TNF-a; 0.95 ± 0.43, P<0.05). In study group, the COX-2 protein in stromal cells cultured with TCDD and TCDD plus TNF-a was significantly increased. (control; 0.81 ± 0.26, TCDD; 1.04 ± 0.38, TCDD plus TNF-a; 1.30 ± 0.90, P<0.05). And the aromatase P450 protein in stromal cells cultured with TNF-a and TCDD and TCDD plus TNF-a was significantly increased (control; 0.76 ± 0.18 vs. TNF-a; 0.93 ± 0.34, TCDD; 0.97 ± 0.41, TCDD plus TNF-a; 1.16 ± 0.41 P<0.05). In the stromal cell cultured with TCDD plus TNF-a than TNF-a and TCDD alone, the aromatase protein was significantly increased (P<0.05). Treatment with the TCDD increased aromatase cytochrome P450 and COX-2 on the eutopic endometrium of patients with and without estrogen dependent diseases, by endometrial stromal cells in culture.

Gonadotropin-releasing hormone agonist and danazol normalize aromatase cytochrome P450 expression in eutopic endometrium from women with endometriosis, adenomyosis, or leiomyomas

Objective To investigate whether GnRH agonists or danazol therapy normalizes estrogen metabolism in the eutopic endometrium of women with endometriosis, adenomyosis, or leiomyomas. Design Prospective clinical study. Setting University hospital. Patient(s) Fifty-three women with endometriosis, adenomyosis, or leiomyomas. Intervention(s) Patients received GnRH agonist or danazol. Biopsy samples of the endometrium were obtained before and after endocrine therapy. Nontreated endometrial explants were cultured in the presence of either drug. Main outcome measure(s) Reverse transcription polymerase chain reaction-Southern blot and immunohistochemical analyses of the endometrial expression of aromatase cytochrome P450, estrogen receptor, progesterone receptor, and Ki-67. Nontreated endometrial explants were cultured in the presence of either drug. Result(s) Messenger RNA and protein of aromatase cytochrome P450 were greatly reduced in the eutopic endometrium of patients treated with GnRH agonist for 2 months or more or with danazol for 1 month or more. Culture of endometrial explants with GnRH agonist (10 −9–10 −7 M) did not change the amount of aromatase cytochrome P450, whereas danazol (10 −7–10 −6 M) efficiently reduced aromatase cytochrome P450 expression. Conclusion(s) Therapy with GnRH agonist or danazol decreases expression of aromatase cytochrome P450 in diseased eutopic endometrium. Endocrine therapy normalized in part the impaired hormonal expression of the eutopic endometrium. GnRH agonist reduced aromatase cytochrome P450 expression mainly by promoting a hypoestrogenic state, whereas danazol reduced aromatase cytochrome P450 in part by direct action on the eutopic endometrium.

Detection of aromatase cytochrome P-450 in endometrial biopsy specimens as a diagnostic test for endometriosis

Objective: To evaluate the clinical usefulness of examining endometrial biopsy specimens for aromatase cytochrome P-450 as a diagnostic test for endometriosis. Design: Retrospective, case-controlled study. Setting: Department of Obstetrics and Gynecology, Kyoto Prefectural University of Medicine, Kyoto, Japan. Patient(s): One hundred five women of reproductive age with normal menstrual cycles underwent endometrial biopsy laparotomy or laparoscopy, and examination of their tissue revealed endometriosis, adenomyosis, and/or leiomyomas. Patients who had cervical carcinoma in situ but no other gynecologic disease were considered to be disease-free. Intervention(s): Endometrial biopsy specimens were collected. Main Outcome Measure(s): The expression of aromatase cytochrome P-450 was examined by reverse transcription-polymerase chain reaction and immunohistochemical analysis. The distribution and intensity of the immunostaining was assessed using a semiquantitative index designed H-score. Result(s): Immunostaining for aromatase cytochrome P-450 was detected in biopsy specimens obtained from patients with endometriosis, adenomyosis, and/or leiomyomas but not in specimens obtained from disease-free patients (H-score <20), with a sensitivity and specificity of 91% and 100%, respectively. Conclusion(s): The expression of aromatase cytochrome P-450 in biopsy specimens of eutopic endometrium distinguishes between disease-free women and women with endometriosis, adenomyosis, and/or leiomyomas. This technique can be used at outpatient infertility clinics as an initial screening procedure to rule out the presence of estrogen-dependent disease.

Expression of aromatase cytochrome P450 in rat H540 leydig tumor cells

The rat H540 Leydig cell tumor has been shown to express cholesterol side-chain cleavage and 17α-hydroxylase cytochrome P450s, 3β-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase, and steroid 5α-reductase, making it a useful model in which to study steroidogenesis. In the current studies, we report that cultured H540 cells express high levels of aromatase cytochrome P450 (P450 arom), which converts androgens to estrogens. Levels of aromatase activity varied from 9.4 to 51.7 pmol/h/mg protein and inhibition of 5α-reductase with finasteride did not significantly effect aromatase measurements, indicating that 5α-reductase is not competing for the substrate used in the aromatase assays. Aromatase activity was decreased 95% by preincubating the cells with 4-hydroxyandrostenedione, an aromatase inhibitor. Characterization of the aromatase mRNA expressed in the H540 cell line demonstrates that, like R2C cells and rat ovarian tissue, three distinct P450 arom mRNA species are detected by Northern analysis, and that these transcripts are derived from the same site of transcription initiation. Despite these similarities, the regulation of aromatase activity by 8-bromo-cAMP in H540 cells differs from both R2C cells and rat ovarian tissue. As the H540 and R2C cell lines appear to have distinct origins, H540 is the second rat Leydig tumor cell line characterized that constitutively expresses high levels of aromatase.

A review of brain aromatase cytochrome P450

Aromatase cytochrome P450 (P450 AROM ) enzyme activity catalyzes the conversion of androgens to estrogens in specific brains areas. During central nervous system development local estrogen formation influences the sexual differentiation of neural structures (i.e., by increasing neurite growth and establishing neural circuitry) and modulates neuroendocrine/reproductive functions and sexual behavior. More than 20 years ago, in 1970, Naftolin et al. provided preliminary direct evidence for the aromatization of androgens by central neuroendocrine tissues. This work created the foundation for the brain aromatase hypothesis. A review of past and recent data reveals the importance of brain aromatase in the development and function of the central nervous system. This review re-examines the aromatase hypothesis in light of recent data and a theoretical proposal is presented in reference to the aromatase mechanism. The metabolic pathway of androgen metabolism by the aromatase cytochrome P450 pathway, cell type, distribution, developmental profile, and regulation of brain aromatase is also presented. The complex nature of brain aromatase is exemplified by recent molecular biology studies examining the expression of aromatase cytochrome P450 during prenatal/postnatal development. Data derived from these studies provide insight into the regulation of the brain aromatase cytochrome P450 gene and suggest an additional level of control for the expression of brain aromatase. These findings present evidence for the utilization of alternative promoter(s) in man and rodents in driving aromatase gene expression in brain. It is clear that molecular mechanism(s) account for the diverse expression of aromatase in different neural tissue sites and during various physiological states or developmental periods. Therefore, further study is necessary in order to understand the significance of the regulation of local estrogen biosynthesis by the aromatase cytochrome P450 gene during prenatal and postnatal development due to the dramatic impact these estrogen molecules have on neural development and their influence on reproductive function and behavior.

Maintenance and regulation of 17 alpha-hydroxylase expression by bovine thecal cells in primary culture.

We have developed and characterized a primary cell culture system to study the regulation of 17 alpha-hydroxylase cytochrome P450 (P45017 alpha) gene expression in bovine thecal cells. Conditions have been established for the dispersal and growth of thecal cells isolated from bovine follicles, which maintain the expression of P45017 alpha for up to 8 days. Bovine theca interna cells were grown to subconfluence and transferred into medium containing forskolin, a stimulator of adenylate cyclase. Levels of P45017 alpha transcripts reached a maximum value after 48 h of stimulation with forskolin. Added progesterone was converted to 17 alpha-hydroxyprogesterone at a rate of 214 pmol/mg protein.h in cells treated with forskolin for 72 h, whereas in control cells, the rate was 9.2 pmol/mg protein.h after 72 h. This was reflected in a 10-fold increase in endogenous androstenedione production by forskolin-stimulated cells. Studies employing various growth factors suggest that transforming growth factor-beta, but not basic fibroblast growth factor, is a potent inhibitor of forskolin-induced 17 alpha-hydroxylase activity and androstenedione production in these cells. We have also characterized this cell culture system with respect to expression of other steroidogenic enzymes. Cholesterol side-chain cleavage cytochrome P450 and 3 beta-hydroxysteroid dehydrogenase transcripts as well as endogenous progesterone accumulation were increased in response to forskolin stimulation. On the other hand, aromatase cytochrome P450 expression was undetectable. The ability to maintain bovine thecal cells, which retain 17 alpha-hydroxylase activity, in culture will provide a model system to study the regulation of expression of the P45017 alpha gene in the bovine ovary.

Relative expression of aromatase cytochrome P450 in human fetal tissues as determined by competitive polymerase chain reaction amplification

Relative expression of aromatase cytochrome P450 in human fetal tissues as determined by competitive polymerase chain reaction amplification

Expression of Aromatase Cytochrome P-450 in Premenopausal and Postmenopausal Human Ovaries: an Immunocytochemical Study*

The cellular distribution of the aromatase cytochrome P-450 enzyme in human ovaries has been investigated immunocytochemically, using an aromatase-specific monoclonal antibody. Ovaries of females ranging in age from prepubertal infant girl through to postmenopausal adulthood were obtained from immediate autopsy or after surgery. The results have revealed temporal and spatial changes in expression of aromatase at different stages of development. No immunoreactive aromatase was detected in the ovary of the 2.5 month infant. In premenopausal ovaries, aromatase was absent from the stromal compartment, but in follicles, a consistent pattern in expression of aromatase was observed, related to their size and developmental stage. Aromatase was not expressed in primordial, primary, or small secondary follicles less than 250 microns diameter. In slightly larger follicles (250-700 microns diameter) aromatase was first detected in a few thecal cells (TC). In more developed secondary through to large preovulatory follicles (greater than 1 cm) TC aromatase immunostain increased in intensity and number of positive cells, and the reaction was localized to a band of theca interna (TI) cells at the TI/theca externa interface. In granulosa cells (GC), aromatase was first detected in follicles in the initial stages of antrum formation (greater than 700 microns), and staining intensified as follicle diameter and antral cavity increased, being maximal in preovulatory follicles. GC aromatase was always found in the presence of TI immunostain. These two cell populations were separated by an unstained layer of TI cells giving the follicle walls a banded appearance. Immunostain was most intense in mural GC, was weaker in antral GC cells and was absent from the cumulus GC. Immunoreactive aromatase was also detected in functional corpora lutea (CL) but was absent from involuting CL's and corpora albicans. Our findings indicate that the immunostained cells of the CL are comprised of the former GC and possibly a subpopulation of former TI cells. In perimenopausal ovaries there was no evidence of any follicular or stromal aromatase immunostain. In postmenopausal ovaries no follicles were observed, but individual cells and clusters of cells in the stromal compartment of 3/7 specimens were found to have an aromatase immunostain reaction. In all cases, the aromatase immunostain reaction was cytoplasmic. The results provide the first direct evidence of the existence of TC aromatase, and of stromal cell aromatase in postmenopausal women.