Résumé
This study aims to examine the effect of superfine powder and aqueous extract of Polygonati Rhizomaon on natural perimenopausal syndrome in rats and explore the underlying mechanism. To be specific, a total of 60 female SD rats(14-15 months old) with estrous cycle disorder were screened by the vaginal smear and randomized into model control group, β-estradiol 3-benzoate group(0.1 mg·kg~(-1)), superfine powder of Polygonati Rhizoma group(0.25, 0.5 g·kg~(-1)) and aqueous extract of Polygonati Rhizoma group(0.25, 0.5 g·kg~(-1)), and another 10 female SD rats(14-15 months old) were selected as the youth control group. The administration lasted 6 weeks. Then the perimenopausal syndrome-related indexes such as body temperature, microcirculatory blood flow of face and ear, vertigo period, salivary secretion, grip force, and bone strength were determined and open field test was conducted. The immune system-related indexes such as the wet weight and index of thymus and spleen, percentage of T lymphocytes and subgroups in peripheral blood, and hematological indexes were measured. In addition, the ovary-related indexes such as estrous cycle, the wet weight and index of uterus and ovary, ovarian tissue morphology, and cell apoptosis were determined. Moreover, hypothalamus-pituitary-ovary axis(HPO)-related indexes such as serum sex hormone levels, cytochrome P450 family 11 subfamily A member 1(CYP11A1), cytochrome P450 family 19 subfamily A member 1(CYP19A1), and cytochrome P450 family 17 subfamily A member 1(P450 17A1) in ovarian tissue were measured. The results showed that the superfine powder and aqueous extract of Polygonati Rhizoma significantly decreased body temperature(anal, facial and dorsal temperature), microcirculatory blood flow in the ear, and vertigo period, increased salivary secretion, grip force, bone strength, total distance and total speed in the open field test, wet weight and index of thymus and spleen, lymphocyte ratio, CD3~+ level, and CD4~+/CD8~+ ratio, reduced neutrophil number and ratio, estrous cycle disorder ratio, and number of ovarian apoptotic cells, raised wet weight and index of uterus, wet weight of ovary, levels of inhibin B(INHB), estradiol(E_2), anti-müllerian hormone(AMH), and ovarian CYP11A1 and CYP19A1, decreased follicle-stimulating hormone(FSH) and luteinizing hormone(LH) content, and improved ovarian tissue morphology. It is suggested that the superfine powder and aqueous extract of Polygonati Rhizoma can improve the symptoms associated with natural perimenopausal syndrome in rats and enhance ovarian function and immune function. The mechanism is that they regulate HPO axis function by increasing estrogen synthesis.
Sujets)
Femelle , Animaux , Rats , Rat Sprague-Dawley , Microcirculation , Cholesterol side-chain cleavage enzyme , Périménopause , Poudres , Cytochrome P-450 CYP1A1Résumé
<p><b>Objective</b>To explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HUMSCs) to differentiate into Leydig cells in the interstitial tissue of the rat testis.</p><p><b>METHODS</b>HUMSCs were obtained by tissue blocks culture attachment and their purity and multi-lineage differentiation ability were verified by flow cytometry and chondrogenic/adipogenic/osteogenic differentiation. Then the HUMSCs were marked by CM-Dil and transplanted into the interstitial tissue of the rat testis. At 4 and 8 weeks after transplantation, the survival and differentiation status of the HUMSCs were observed by immunofluorescence staining and flow cytometry. The suspension of the rat Leydig cells was obtained at 8 weeks for determining the expression of the Leydig cell marker 3β-HSD in the HUMSCs, the cells labeled with CM-Dil were sorted and cultured, and the medium collected after 3 days of culture for measurement of the testosterone level.</p><p><b>RESULTS</b>The expression of the Leydig cell marker CYPllal was not observed in the HUMSCs at 4 weeks but found at 8 weeks after transplantation and the differentiation rate of 3β-HSD was about 14.5% at 8 weeks. CM-Dil labeled cells survived after sorting and testosterone was detected in the medium.</p><p><b>CONCLUSIONS</b>HUMSCs are likely to differentiate into Leydig cells in the interstitium of the rat testis.</p>
Sujets)
Animaux , Humains , Mâle , Rats , Marqueurs biologiques , Métabolisme , Carbocyanines , Différenciation cellulaire , Cholesterol side-chain cleavage enzyme , Métabolisme , Études de faisabilité , Cellules de Leydig , Biologie cellulaire , Métabolisme , Cellules souches mésenchymateuses , Biologie cellulaire , Testicule , Biologie cellulaire , Facteurs temps , Cordon ombilical , Biologie cellulaireRésumé
Tribulus terrestris has been commonly used in folk medicine to energize, vitalize and improve sexual function and physical performance in men and laboratory rats. To study the effect of Tribulus terrestris on the number of Leydig cells. Tribulus terrestris was given to mature male rats as an oral single herbal suspension in a dose of 2.0mg /1000gbody weight for 14 days to stimulate spermatogenesis. Formalin fixed paraffin-embedded tissue sections were performed for histological, immunohistochemical and morphometrical studies. Histological study revealed wider seminiferous tubules and increased spermatocytes population with an increased sperm density inside the lumen of the tubules. Morphometrically, the diameters of seminiferous tubules and thickness of the germinal epithelia were significantly increased in Tribulus terrestris treated rats than that of the control group. There was no significant difference between the number of Leydig cells in the control and experimental groups. The activity of Leydig cells, manifested by the increments in the diameters, thickness of germinal epithelia and the density of the sperms inside seminiferous tubules, was increased but their number remain unaffected in spite of using the aphrodisiac agent, Tribulus terrestris
Sujets)
Tribulus , Phytothérapie , Cholesterol side-chain cleavage enzyme , Rats , Immunohistochimie , TesticuleRésumé
<p><b>OBJECTIVE</b>To establish an in vitro model of cultured mouse testis using rotary aerobic culture.</p><p><b>METHODS</b>Rotary aerobic incubation with optimized culture conditions was used for in vitro culture of mouse testis, and the morphology of the cultured testicular tissues was compared with that cultured in Transwell chambers. The changes in the testicular tissue structure were examined using HE staining, and the cell proliferation was assessed with BrdU staining. Testosterone concentrations in the culture medium were tested with radioimmunoassay and the expression of the functionally related proteins in the testis was detected using immunohistochemistry.</p><p><b>RESULTS</b>The testicular tissue cultured by optimized rotary aerobic culture presented with more intact histological structure with the size of the testis ranged from 0.3 to 0.8 mm(3). In the two culture systems, the prolifeation index of the spermatogonia increased and that of Sertoli cells decreased with time, and such changes in spermatogonia and Sertoli cell proliferation indices became statistically significant at 3 days (P<0.05) and 5 days (P<0.05) of culture, respectively, as compared with those at 1 day. The concentration of testoerone in the culture media decreased significantly with incubation time (P<0.05). At 3 days of culture, the protein expression of 3β-hydroxysteroid dehydrogenase, cytochrome P450 17α-hydroxylase and cholesterol side-chain cleavage enzyme was detected in Leydig cell cytoplasm and vimentin expression in Sertoli cell cytoplasm.</p><p><b>CONCLUSION</b>An in vitro model of cultured mouse testis has been successfully established using rotary aerobic incubation.</p>
Sujets)
Animaux , Mâle , Souris , 17-Hydroxysteroid dehydrogenases , Métabolisme , Cholesterol side-chain cleavage enzyme , Métabolisme , Milieux de culture , Chimie , Cellules de Leydig , Biologie cellulaire , Techniques de culture d'organes , Dosage radioimmunologique , Cellules de Sertoli , Biologie cellulaire , Spermatogonies , Biologie cellulaire , Testicule , Testostérone , Chimie , Vimentine , MétabolismeRésumé
Congenital lipoid adrenal hyperplasia (lipoid CAH) is the most fatal form of CAH, as it disrupts adrenal and gonadal steroidogenesis. Most cases of lipoid CAH are caused by recessive mutations in the gene encoding steroidogenic acute regulatory protein (StAR). Affected patients typically present with signs of severe adrenal failure in early infancy and 46,XY genetic males are phenotypic females due to disrupted testicular androgen secretion. The StAR p.Q258X mutation accounts for about 70% of affected alleles in most patients of Japanese and Korean ancestry. However, it is more prevalent (92.3%) in the Korean population. Recently, some patients have been showed that they had late and mild clinical findings. These cases and studies constitute a new entity of 'nonclassic lipoid CAH'. The cholesterol side-chain cleavage enzyme, P450scc (CYP11A1), plays an essential role converting cholesterol to pregnenolone. Although progesterone production from the fetally derived placenta is necessary to maintain a pregnancy to term, some patients with P450scc mutations have recently been reported. P450scc mutations can also cause lipoid CAH and establish a recently recognized human endocrine disorder.
Sujets)
Femelle , Humains , Mâle , Grossesse , Allèles , Asiatiques , Cholestérol , Cholesterol side-chain cleavage enzyme , Gonades , Hyperplasie , Placenta , Prégnénolone , ProgestéroneRésumé
Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.
Sujets)
Animaux , Bovins , Femelle , Milieux de culture/pharmacologie , Oestradiol/pharmacologie , Follicule ovarique/effets des médicaments et des substances chimiques , Progestérone/pharmacologie , Techniques de culture de tissus , Analyse de variance , Aromatase/génétique , Milieux de culture sans sérum , Cholesterol side-chain cleavage enzyme/génétique , Expression des gènes , Follicule ovarique/anatomie et histologie , Phosphoprotéines/génétique , Progesterone reductase/génétique , RT-PCR , Récepteur FSH/génétique , /génétiqueRésumé
<p><b>OBJECTIVE</b>To explore the effects of di-butyl phthalate (DBP) on the reproductive system of adolescent male rats.</p><p><b>METHODS</b>Sprague-Dawley (SD) rats aged 5 weeks were assigned to receive corn oil (vehicle control) or DBP orally at 10, 100 and 500 mg/(kg x d) for 30 days. After the exposure, the testis, epididymis, liver and pituitary of the rats were weighted and their ratios to the body weight obtained. Histopathological changes of the testis and epididymis were examined by Hematoxylin-eosin staining, the levels of testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the serum were measured by radioimmunoassay, and the relative mRNA expressions of the steroidogenesis acute regulatory protein (StAR), proliferating cell nuclear antigen (PCNA), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc) and scavenger receptor (SR) were detected by real-time quantitative RT-PCR.</p><p><b>RESULTS</b>DBP induced significant histopathological changes in the testicular tissue at 100 and 500 mg/(kg x d), and decreased the testicular and epididymal weights, inhibited the mRNA expressions of StAR and PCNA, reduced the levels of T and LH, and elevated the level of FSH at 500 mg/(kg x d). At the dose of 10 mg/(kg x d), DBP increased serum LH and FSH and the mRNA expression of P450scc. While the SR mRNA expression showed no significant changes in all the groups.</p><p><b>CONCLUSION</b>High level of DBP has apparent toxic effect on reproductive system of male rats. Low - dose DBP can increase the level of serum gonadotropin LH and affect the mRNA expression of P450scc in the testis.</p>
Sujets)
Animaux , Mâle , Rats , Cholesterol side-chain cleavage enzyme , Métabolisme , Phtalate de dibutyle , Toxicité , Hormone folliculostimulante , Sang , Hormone lutéinisante , Sang , Phosphoprotéines , Métabolisme , Antigène nucléaire de prolifération cellulaire , Métabolisme , ARN messager , Génétique , Rat Sprague-Dawley , Récepteurs éboueurs , Métabolisme , Testicule , MétabolismeRésumé
El síndrome de poliquistosis ovárica (PCOS) es un desorden endocrino-metabólico de naturaleza multifactorial, con una marcada predisposición genética, que afecta al 6% de las mujeres en edad reproductiva. Se caracteriza por la presencia de hiperandrogenismo, oligo-anovulación y ovarios poliquísticos. Entre los genes candidatos se encuentran aquellos que codifican para enzimas que actúan en la síntesis de andrógenos. Dos de los genes candidatos son el CYP17 y el CYP11alfa que codifican para la 17alfa hidroxilasa (P45017alfa) y para el P450scc (colesterol side chain cleavage) respectivamente. Los polimorfismos en estos genes están asociados al desarrollo del fenotipo hiperandrogénico. Nuestro objetivo fue analizar las frecuencias alélicas de los polimorfismos de los dos genes mencionados en población con PCOS, compararla con población normal y analizar la relación de cada variante alélica con el fenotipo hiperandrogénico correspondiente. Se analizaron 65 pacientes y 58 controles sanos en los que se determinaron niveles de testosterona y frecuencia de polimorfismos en los genes mencionados. Se observó una diferencia estadísticamente significativa cuando se asoció el grupo de mayor nivel de androgenemia con la presencia del genotipo A2/A2 del gen CYP17, y se hallaron mayores niveles de andrógenos circulantes en las pacientes con PCOS portadoras del alelo 216- del gen CYP11alfa. Nuestros resultados sugieren que ambos alelos juegan un rol menor en el desarrollo de PCOS y podrían ser considerados como potenciales marcadores de riesgo genético para el desarrollo del fenotipo hiperandrogénico.
The polycystic ovary syndrome (PCOS) is a heterogeneous multifactorial endocrine metabolic disorder with genetic predisposition affecting 6% of women in the reproductive age. This syndrome is characterized by the presence of oligo-anovulation, hyperandrogenism and polycystic ovaries. Several genes have been postulated as responsible for the etiology of this disorder. Among these genes are those encoding the enzymes involved in the ovarian androgen biosynthesis. Two of the candidate genes are the CYP17 and the CYP11alpha, encoding the 17-alpha-hydroxylase (P45017alpha) and the cholesterol side chain cleavage (P450scc) respectively. The polymorphisms of these genes are linked to the development of an hyperandrogenic phenotype. The aim of this work was to analyze the allelic frequencies of such polymorphisms in a cohort of women with PCOS and to compare them with those of healthy women. Furthermore, the correlation between each allelic variant and the corresponding hyperandrogenic phenotype was also assessed. Therefore, 65 patients and 58 age matched healthy controls were analyzed. The serum levels of testosterone and the frequency of each polymorphism were determined. When the PCOS population was analyzed, a significant statistical difference was found when relating the group with the highest androgenemia level with the presence of A2/A2 genotype of CYP 17 gene, and a higher level of circulating androgen was found in PCO women carrying the 216- allele of CYP11alpha gene (that did not reach statistical significance). Our results suggest that both alleles play a minor role in the development of PCOS and could be a genetic risk marker of the hyperandrogenic phenotype.
Sujets)
Femelle , Humains , Cholesterol side-chain cleavage enzyme/génétique , Hyperandrogénie/génétique , Syndrome des ovaires polykystiques/génétique , Polymorphisme génétique/génétique , /génétique , Androgènes/analyse , Androgènes/pharmacocinétique , Biodisponibilité , Études cas-témoins , Marqueurs génétiques/génétique , Réaction de polymérisation en chaîne , Polymorphisme de restriction , Testostérone/analyse , Testostérone/pharmacocinétiqueRésumé
<p><b>AIM</b>To study the effect and mechanism of gonadotrophin-releasing hormone (GnRH) on murine Leydig cell steroidogenesis.</p><p><b>METHODS</b>Purified murine Leydig cells were treated with GnRH-I and -II agonists, and testosterone production and steroidogenic enzyme expressions were determined.</p><p><b>RESULTS</b>GnRH-I and -II agonists significantly stimulated murine Leydig cell steroidogenesis 60%-80% in a dose- and time-dependent manner (P < 0.05). The mRNA expressions of steroidogenic acute regulatory (StAR) protein, P450scc, 3beta-hydroxysteroid dehydrogenase (HSD), but not 17alpha-hydroxylase or 17beta-HSD, were significantly stimulated by both GnRH agonists with a 1.5- to 3-fold increase (P < 0.05). However, only 3beta-HSD protein expression was induced by both GnRH agonists, with a 1.6- to 2-fold increase (P < 0.05).</p><p><b>CONCLUSION</b>GnRH directly stimulated murine Leydig cell steroidogenesis by activating 3b-HSD enzyme expression.</p>
Sujets)
Animaux , Mâle , Souris , 3-Hydroxysteroid dehydrogenases , Génétique , Technique de Western , Séparation cellulaire , Cellules cultivées , Cholesterol side-chain cleavage enzyme , Relation dose-effet des médicaments , Hormone de libération des gonadotrophines , Pharmacologie , Cellules de Leydig , Métabolisme , Souris de lignée C57BL , Phosphoprotéines , Génétique , ARN , RT-PCR , Maturation sexuelle , Physiologie , Stéroïdes , TestostéroneRésumé
<p><b>OBJECTIVE</b>To investigate the effects of Ginkgo biloba extract (EGB) on the testosterone synthesis in the Leydig cells of type 2 diabetic rats.</p><p><b>METHODS</b>Thirty male SD rats were equally randomised into a normal control, a type 2 diabetic and an EGB group. Morphological changes of Leydig cells were observed by light microscopy (LM) and transmission electron microscopy (TEM), concentrations of serum luteinizing hormone (LH) and testosterone (T) were determined by enzyme linked immunosorbent assay (ELISA), and the mRNA levels in the steroidogenic acute regulatory protein (StAR), cytochrome P450 side chain cleavage (P450scc), cytochrome P450 17a-hydroxylase (P450c17), 17beta-hydroxysteroid dehydrogenase 3 (17beta-HSD3) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD1) from the Leydig cells were examined by RT-PCR.</p><p><b>RESULTS</b>Compared with the normal control, there was a significant decrease in the number and volume of Leydig cells, the levels of serum LH and T and the expression of mRNA in StAR, P450scc, 17beta-HSD3 and 3beta-HSD1 in the type 2 diabetes group. And the expression of the P450c17 gene showed a tendency of descending, but with no significance. Compared with the type 2 diabetes group, 12 weeks of EGB treatment caused very slight pathological changes in the Leydig cells, significantly increased the concentrations of blood LH and T, markedly elevated the levels of mRNA in StAR and P450scc and induced an ascending tendency of the expressions of P450c17, 17beta-HSD3 and 3beta-HSD1.</p><p><b>CONCLUSION</b>EGB enhances testosterone synthesis and secretion of Leydig cells by reducing the impairment of the testis in type 2 diabetic rats.</p>
Sujets)
Animaux , Mâle , Rats , 17-Hydroxysteroid dehydrogenases , Génétique , Cholesterol side-chain cleavage enzyme , Génétique , Diabète de type 2 , Sang , Génétique , Test ELISA , Expression des gènes , Ginkgo biloba , Chimie , Hydroxysteroid dehydrogenases , Génétique , Cellules de Leydig , Métabolisme , Hormone lutéinisante , Sang , Microscopie électronique à transmission , Phosphoprotéines , Génétique , Extraits de plantes , Pharmacologie , ARN messager , Génétique , Métabolisme , Rat Sprague-Dawley , RT-PCR , Testostérone , SangRésumé
The steroidogenic acute regulatory (StAR) protein is an essential component in the regulation of steroid biosynthesis in adrenal and gonadal cells. The StAR protein has a high tissue specificity, located on the mitochondrial membranes of some relative cells. It regulates the transfer of cholesterin from extracellular into intracellular and plays a dominant role in steroidogenic synthesis. Recent studies have also shown that the transcription and expression of StAR are modulated not only through the cAMP-PKA dependent pathway, but also by multiple hormones and cytokines, which contributes to the regulation of cholesterin synthesis.
Sujets)
Animaux , Humains , Souris , Rats , Cholestérol , Métabolisme , Cholesterol side-chain cleavage enzyme , Métabolisme , Éthistérone , Métabolisme , Régulation de l'expression des gènes , Mitochondries , Métabolisme , Phosphoprotéines , Génétique , PhysiologieRésumé
To determine the (tttta)n repeat polymorphisms at the promoter region of CYP11alpha gene, and study its linkage to hyperandrogenism of polycystic ovary syndrome (PCOS) in Chinese women, a case-control study was conducted in the Reproductive Medical Center of the Second Affiliated Hospital of Zhengzhou University (Zhengzhou, China). 96 PCOS patients and 78 healthy control women were included. CYP11alpha (tttta)n repeat-polymorphism genotyping analysis was performed by using polymerase chain reaction (PCR). Serum pituitary hormone and total testosterone levels were measured by ELISA. 4 different CYP11alpha (tttta)n allelles were identified, corresponding to 4-, 6-, 8-, and 9-repeat-unit alleles. The frequency and distribution of these alleles are 0.16, 0.33, 0.38, and 0.13 respectively in PCOS patients, as compared with 0.20, 0.34, 0.35, and 0.11 respectively in healthy controls. There were no significant differences between these two groups. Moreover, no correlation between the polymorphism of CYP11alpha gene and serum testosterone level of patients with PCOS and controls was observed. It is concluded that microsatellite polymorphism (tttta)n of gene CYP11alpha exists in Chinese women and the polymorphism of CYP11alpha gene does not play an important role in the pathogenesis of Chinese patients with PCOS, especially in patients with hyperandrogenism.
Sujets)
Cholesterol side-chain cleavage enzyme/génétique , Hyperandrogénie/complications , Hyperandrogénie/génétique , Répétitions microsatellites , Syndrome des ovaires polykystiques/complications , Syndrome des ovaires polykystiques/génétique , Polymorphisme génétique/génétiqueRésumé
<p><b>OBJECTIVE</b>To study the influence of electromagnetic irradiation on cytochrome P450 cholesterol side chain lyase (P450scc) in adult rat testis tissues and to assess the protective effect of the copper shield.</p><p><b>METHODS</b>Healthy male Wistar rats were randomized into a control group, an electromagnetic irradiation group and a wholly shielded group (with the copper shielding net). The electromagnetic irradiation group and the shielded group were set for 4 phases of 3, 6, 24 and 72 hours after irradiation, 15 rats for each phrase. The testosterone contents in the serum of the irradiated rats at 3, 6, 24 and 72 hours and in that of the controls were measured by radioimmunoassay(RIA), and so was the level of the P450scc mRNA in the testis tissues by semi-quantitative RT-PCR. And the effect of the copper shielding net on testosterone and P450scc mRNA was observed.</p><p><b>RESULTS</b>The contents of testosterone and the P450scc mRNA level in the irradiated group were significantly lower than in the control rats, decreased by 83.9% and 56.9% at 3 hours (P < 0.01), 54.8% and 27.3% at 6 hours (P < 0.01), restored to normal at 24 hours, but again reduced by 60.1% and 56.1% respectively (P < 0.01). While in the shielded group, no significant change was observed either in the testosterone of the serum or in the P450scc mRNA expression in the testis tissues.</p><p><b>CONCLUSION</b>Electromagnetic irradiation may affect the transcription of P450scc in adult rat Leydig cells and thereby decrease the testosterone synthesis. Whole-body shielding with the copper net may achieve satisfactory effect.</p>
Sujets)
Animaux , Mâle , Rats , Cholesterol side-chain cleavage enzyme , Génétique , Cuivre , ARN messager , Génétique , Radioprotection , Dosage radioimmunologique , Répartition aléatoire , Rat Wistar , RT-PCR , Testicule , Métabolisme , Effets des rayonnements , Testostérone , SangRésumé
The enzyme complex 3b-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. The expression of 3beta-HSD in human uterine endometrium during the menstrual cycle and decidua was examined in an effort to understand its role during ova implantation. 3beta-HSD was weakly expressed in the glandular epithelium of the proliferative phase and moderately expressed in the glandular epithelium of secretory phase of the endometrium. In the decidua of the ectopic pregnancy, 3beta-HSD was strongly expressed. The human uterine endometrial 3beta-HSD was identified as being the same type as the placental 3beta-HSD by RT-PCR and sequence analysis. In addition to the expression of 3beta-HSD, P450scc was expressed in the decidua of the ectopic pregnancy. These results suggest that pregnenolone might be synthesized from cholesterol by P450scc de novo and then, it is converted to progesterone by 3beta-HSD in the uterine endometrium. The data implies that the endometrial 3beta-HSD can use not only the out-coming pregnenolone from the adrenal gland but also the self- made pregnenolone to produce progesterone. The de novo synthesis of progesterone in the endometrium might be a crucial factor for implantation and maintenance of pregnancy.
Sujets)
Femelle , Humains , Grossesse , Cholestérol/composition chimique , Cholesterol side-chain cleavage enzyme/biosynthèse , Caduques/enzymologie , Endomètre/enzymologie , Expression des gènes/physiologie , Cycle menstruel/physiologie , Complexes multienzymatiques/biosynthèse , Placenta/enzymologie , Prégnénolone/biosynthèse , Progestérone/biosynthèse , Progesterone reductase/biosynthèse , Steroid isomerases/biosynthèseRésumé
PURPOSE: A crucial event in the acute regulation of steroidogenesis by tropic hormone is the delivery of cholesterol from cytosol into the mitochondrial inner membrane where it is converted to pregnenolone by the cytochrome P45O cholesterol side chain cleavage enzyme. Recently, it has been observed that the acute production of steroid hormone depends on a rapidly synthesized, cycloheximide sensitive and highly labile protein that appeared in response to tropic hormone. This protein was named as Steroidogenic Acute Regulatory(StAR) protein. The purpose of this experiment was to detect the StAR protein in human testicular tissue. MATERIALS AND METHODS: Human testicular tissues were obtained during the surgery in patients with cryptorchidism, infertility, and prostate cancer Northern blotting hyridization and RT-PCR were performed to detect the StAR mRNA in human testicular tissues. In addition, immunohistochemical staining was performed to test the presence of the StAR protein in human testicular tissues. RESULTS: The StAR mRNA was detected as a weak band in prostate cancer and infertility patients, but not detected in cryptorchidism patients by Northern blotting hybridization. In RT-PCR of human testicular tissues for StAR mRNA, RT-PCR products(about 2000p) were shown to have in cryptorchidism, infertility, and prostate cancer patients. In immunohistochemical staining of human StAR protein, immunoreactivity of human StAR protein was positive in the interstitial tissues of human testis. CONCLUSIONS: The StAR protein that plays a key role in the steroidogenesis was detected in human testicular tissues and this protein will be effective in pathogenesis and treatment of the diseases that are associated with steroid hormones.
Sujets)
Humains , Mâle , Technique de Northern , Cholestérol , Cholesterol side-chain cleavage enzyme , Cryptorchidie , Cycloheximide , Cytochromes , Cytosol , Infertilité , Membranes , Prégnénolone , Tumeurs de la prostate , ARN messager , TesticuleRésumé
Se reporta una vía alternativa para la síntesis del 3-etilencetal-androsta-3, 17-diona, el cual es un intermedio importante para la obtención de corticoides por construcción de la cadena lateral de 17-ceto esteroides. En nuestro caso, partiendo de la androsta-4-ene-3, 17-diona, se obtuvo el producto deseado en 3 pasos de síntesis con buenos rendimientos
Sujets)
Androstérone/antagonistes et inhibiteurs , Androstérone/synthèse chimique , Cholesterol side-chain cleavage enzyme , Voie alterne d'activation du complémentRésumé
Familial male pseudohermaphroditism (MPH) due to 17,20-desmolase deficiency is rare. Here we present two siblings with MPH possibly due to 17,20-desmolase deficiency. The first patient presented with unambiguous female external genitalia and hypergonadotrophic hypogonadism. Chromosomal analysis revealed 46 XY. Ultrasound evaluation of pelvis revealed gonads in the inguinal canal, and no uterus. These findings were confirmed on laparotomy. Histology revealed the gonads to be testes. The second patient had ambiguous genitalia (perineoscrotal hypospadias, bifid scrotum with palpable gonads) with a 46 XY chromosomal pattern. Both patients had high plasma 17-hydroxy progestrone (17 OHP), low normal dehydro epiandrosterone sulphate (DHEAS) and low plasma testosterone. Plasma testosterone and DHEAS showed no response to ACTH or HCG. These features are compatible with the diagnosis of 17,20-desmolase deficiency.
Sujets)
Adolescent , Adulte , Cholesterol side-chain cleavage enzyme/déficit , Femelle , Études de suivi , Humains , Hydrocortisone/analyse , Mâle , Troubles du développement sexuel/enzymologie , Testostérone/analyse , Résultat thérapeutiqueRésumé
A male infant presented with hyponatremia, hyperkalemia, generalized skin hyperpigmentation, and female type external genitalia. These clinical findings were compatible with mineralocorticoid, glucocorticoid and androgen insufficiency. Serum cortisol, progesterone and testosterone levels were extremely low after ACTH stimulation test, suggestive of defect in all of the adrenal steroidogenesis. Computed tomography demonstrated enlarged adrenal glands. The diagnosis of P450scc deficiency or lipoid congenital adrenal hyperplasia was based on all these characteristics. Physiologic replacement therapy with hydrocortisone and 9 alpha-fluorocortisol were effective and the patient achieved normal growth. The clinical characteristics, differential diagnoses, and prenatal diagnosis are discussed and reviewed.
Sujets)
Hyperplasie congénitale des surrénales/diagnostic , Hormone corticotrope/diagnostic , Cholesterol side-chain cleavage enzyme/déficit , Diagnostic différentiel , Femelle , Humains , Nouveau-né , Mâle , Pedigree , ThaïlandeRésumé
1. The objective of the present investigation was to study some of the possible mechanisms involved in the protective effect of sucrose ingestion against liver necrosis induced by acetaminophen. Three groups of male Wistar rats (220-260 g) were submitted to the following experimental conditions for a period of 42 h: free access to a balanced commercial diet (Group I), an exclusive sucrose diet (Group II) and fasting (Group III). At the end of the experiment, hepatic cytochrome P450 levels were measured in 11 rats from each group, plasma antipyrine half-life (t1/2) was determined in 40 rats from each group, and hepatic glutathione (GSH) concentration in 10 rats from each group. GSH consumption elicited by a high dose of acetaminophen (ACP, 1.0 g/kg, by gavage) was also determined in 30 rats each from Groups II and III. 2. The liver of Group II rats presented a significant reduction of cytochrome P450 levels in the microsome fraction (range 0.31-0.46, median, 0.37 nmol/mg vs range 0.60-0.93, median 0.74 for group I, and range 0.63-1.22, median 0.91 for group III, reported as nmol/mg microsome protein; range 23.8-48.4, median 40.4 vs 66.6-130, median 81.8 for group I and range 59.0-117.1, median 77.1 for group III, reported as nmol/100 g body weight), and a prolongation of antipyrine half-life (146.4 vs 83.4 min for group I and 93.6 for group III) when compared with the rats of the two other groups. 3. Since the toxicity of acetaminophen depends on the production of a reactive metabolite by the cytochrome P450 system in the liver, we conclude that changes in this system brought about by exclusive sucrose ingestion for 42 h may explain the liver protection against the toxicity of a high dose of the drug even in the presence of a significant concomitant reduction in liver GSH levels