ABSTRACT
OBJECTIVE: This study sought to examine corticosteroidogenic enzyme activities in normo- and hyperandrogenic polycystic ovary syndrome (PCOS) patients. SUBJECTS AND METHODS: This cohort study included 81 patients with biochemical hyperandrogenism and 41 patients with normal androgen levels. Enzyme activities were assessed according to the serum steroid product/precursor ratios at baseline and after adrenal stimulation. RESULTS: At baseline, in the delta 4 (Δ4) pathway, hyperandrogenic patients showed greater 17-hydroxylase and 17,20 lyase activities in converting progesterone (P4) into 17-hydroxyprogesterone (17-OHP4) and 17-hydroxypregnenolone (17-OHPE) into androstenedione (A) (p = 0.0005 and p = 0.047, respectively) compared to normoandrogenic patients. In the delta 5 (Δ5) pathway, the 17-hydroxylase and 17,20 lyase enzymes showed similar activities in both groups. Hyperandrogenic patients presented lower 21-hydroxylase, lower 11β-hydroxylase (p = 0.0001), and statistically significant increases in 3β-hydroxysteroid dehydrogenase II (3β-HSDII) activities (p < 0.0001). Following tetracosactrin stimulation, only the 17,20 lyase activity remained up-regulated in the Δ4 pathway (p < 0.0001). CONCLUSION: Hyperandrogenic patients had higher 17,20 lyase activity, both at baseline and after adrenal stimulation. Greater conversion of dehydroepiandrosterone (DHEA) into A with normal conversion of 17-OHPE to 17-OHP4 in hyperandrogenic PCOS patients indicated different levels of 3β-HSDII activity in adrenal cells, and hyperandrogenic patients had lower 11β-hydroxylase and 21-hydroxylase activities.
OBJETIVO: O objetivo deste estudo foi examinar a atividade de enzimas responsáveis pela produção de corticosteroides em pacientes normo e hiperandrogênicas com síndrome de ovários policísticos (SOP). SUJEITOS E MÉTODOS: A coorte estudada incluiu 81 pacientes com hiperandrogenismo bioquímico e 41 pacientes com níveis normais de androgênio. A atividade enzimática foi avaliada de acordo com as proporções de produto/precursor do esteroide sérico, no momento inicial do estudo e depois de estimulação adrenal. RESULTADOS: No momento inicial, na via delta 4 (Δ4), as pacientes hiperandrogênicas mostraram maior atividade da 17-hidroxilase e 17,20 liase na conversão da progesterona (P4) em 17-hidroxiprogesterona (17-OHP4) e na conversão da 17-hidroxipregnenolona (17-OHPE) em androstenediona (A) (p = 0,0005 e p = 0,047, respectivamente) em comparação com pacientes normoandrogênicas. Na via delta 5 (Δ5), a 17-hidroxilase e a 17,20 liase mostraram atividades similares nos dois grupos. As pacientes hiperandrogênicas mostraram menor atividade da 21-hidroxilase, menor atividade da 11β-hidroxilase (p = 0,0001) e aumento estatisticamente significativo na atividade da 3β-hidroxiesteroide desidrogenase II (3β-HSDII) (p < 0.0001). Após a estimulação com tetracosactrin, apenas a atividade da 17,20 liase permaneceu regulada para cima na via Δ4 (p < 0.0001). CONCLUSÃO: As pacientes hiperandrogênicas apresentaram atividade mais alta da 17,20 liase, tanto no momento inicial quanto depois da estimulação adrenal. Maior conversão da desidroepiandrosterona (DHEA) em A com conversão normal da 17-OHPE em 17-OHP4 em pacientes hiperandrogênicas com SOP indica níveis diferentes de atividade da 3β-HSDII em células da adrenal, e pacientes hiperandrogênicas apresentaram menores atividades da 11β-hidroxilase e da 21-hidroxilase.
Subject(s)
Adult , Female , Humans , Adrenal Glands/enzymology , Hyperandrogenism/enzymology , Polycystic Ovary Syndrome/enzymology , Steroid Hydroxylases/metabolism , /metabolism , Adrenal Hyperplasia, Congenital/enzymology , Case-Control Studies , Dehydroepiandrosterone/metabolism , Enzyme Activation , Lyases/metabolism , /metabolism , /metabolism , /metabolismABSTRACT
Adult male rats received daily injections (sc) of gonadotropin releasing hormone antagonist (0.2 mg/kg(-1) x day(-1)) for 21 days when they were sacrificed on day 22, adrenal weight, adrenal A5-3beta (delta 5-3beta) hydroxysteroid dehydrogenase (Delta5-3beta-HSD) activity and serum level of corticosterone were increased significantly while testicular 17beta (17beta) hydroxysteroid dehydrogenase (17beta-HSD) activity and serum level of testosterone and spermatogenesis were decreased in the rats fed on 5% casein diet. GnRH antagonist treated rats fed on 20% casein diet, resulted significant decrease in adrenal weight, serum corticosterone and adrenal A5-3beta-HSD activity while testicular 17beta-HSD activity serum testosterone levels and the weights of sex organs were increased with respect to anti GnRH treated rats fed on 5% casein diet. But the GnRH antagonist treated rats fed on 20% casein diet showed decreased spermatogenesis quantitatively and sperm count appeared similar to anti GnRH treated rats fed on 5% casein diet. These results indicate that high casein diet protects adrenocortical activity and stimulates testosterone synthesis without effecting spermatogenic arrest in GnRH antagonist treated rats. It may be concluded that GnRH antagonist in presence of high milk protein diet may be considered to be a suitable antihormone in the development of an ideal male contraceptive.
Subject(s)
Adrenal Glands/enzymology , Animals , Caseins/administration & dosage , Dietary Proteins/administration & dosage , Gonadotropin-Releasing Hormone/antagonists & inhibitors , Hydroxysteroid Dehydrogenases/metabolism , Male , Rats , Rats, Wistar , Spermatogenesis , Testis/enzymologyABSTRACT
La adrenarca se caracteriza por el aumento de secreción de andrógenos adrenales. Ocurre entre los 6-8 años de edad en humanos. Con el objetivo de evaluar el posible rol regulatorio de la 3beta HSD en la síntesis de andrógenos en la adrenal humana, se analizó la abundancia del ARNm de 3beta HSD (Dot blot y RT-PCR semicuantitativa(S)) en 11 tejidos adrenales humanos normales en dos grupos (Gr) de edades, Gr1: <8 años (a) (n= 6, r: 0.1-2.5) y Gr2: si 8a (n = 5, r: 8.0-13.0); en tejido de un tumor adrenal con Síndrome de Cushing (TSC) y de 2 T virilizante (TV) y en células adrenales del TSC y 1 TV al 3er día de cultivo (basal y con ACTH e IGF-1). El ARNm de 3beta HSD fue más alto en el Gr1 que en el Gr2 (Dot blot: 4.65 + 2.70 y 0.28 + 0.27 UA, p = 0.006; RT-PCRS: 21.50 + 12.50 y 6.77 + 3.78 UA, p = 0.039 resp). Por otra parte, en el tejido TSC (8.74 + 1.74) fue más alto que en los TV (0.47 + 0.02, 0.87 + 0.08) p = 0.001. En cultivo del TSC el ARNm basal (0.82 + 0.10) disminuyó con ACTH (0.55 + 0.06), p = 0.005 y se incrementó con IGF-1 (2.36 + 0.07) p = 0.006. En el TV no hubo cambios. Al 3er día de cultivo la DHEAS y androstenediona basales fueron en TV 1170.0 + 210.0 y 335.0 + 29.0, TSC 17.1 + 3.5 y 73.7 + 11.7 pmol/10(6) células en 24 hs respectivamente y se incrementaron bajo ACTH en TV (2006.0 + 360.0 y 525.0 + 76.0) y en TSC 29.8 + 5.4 y 366.8 + 129) p<0.05 y disminuyeron con IGF-1 sólo en TSC (7.9 + 2.4 y 43.7 + 6.1) p<0.05. Estos datos sugieren que la adrenarca humana podría ser secundaria a una disminución de la abundancia del ARNm de 3beta HSD. El hecho de que bajo ACTH aumenta la secreción de andrógenos y disminuye el ARNm de 3beta HSD, mientras que el IGF-1 ejerce un efecto inverso, aporta nuevas evidencias del rol regulador ejercido por la 3beta en la síntesis de andrógenos adrenales.
Subject(s)
Child , Humans , Male , Adolescent , Child, Preschool , Infant , 3-Hydroxysteroid Dehydrogenases/metabolism , Adrenal Gland Neoplasms/enzymology , Adrenal Glands/enzymology , Androgens/metabolism , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/analysis , RNA, Messenger/metabolismABSTRACT
The activities of aspirin (acetylsalicylic acid)-esterases were measured in several tissues (liver, kidney, adrenal glands, brain and serum) from adult male and female Wistar rats. In males, both aspirin-esterase I (assayed at pH 5.5) and II (assayed at pH 7.4) activities were higher in liver homogenates when compared to females (aspirin-esterase I: males 48.9 ñ 4.8 (N = 8) and females 29.3 ñ 4.2 (N = 8) nmol of salicylic acid formed min-1 mg protein-1; aspirin-esterase II: males 41.4 ñ 4.1 (N = 8) and females 26.1 ñ 4.5 (N = 8) nmol of salicylic acid formed min-1 mg protein-1, P<0.001). In serum, enzyme activity was higher in females than in males (aspirin-esterase I: males 0.85 ñ 0.06 (N = 6) and females 1.18 ñ 0.11 (N = 6) nmol of salicylic acid formed min-1 mg protein-1; aspirin-esterase II: males 1.03 ñ 0.13 (N = 6) and females 1.34 ñ 0.11 (N = 6) nmol of salicylic acid formed min-1 mg protein-1, P<0.001). In the other tissues assayed, no statistically significant difference between males and females was found. There were no statistically significant differences when the enzymes were assayed in different phases of the estrous cycle in liver and serum. These results show that the differences in aspirin-esterase activity observed between males and females are not due to the estrous cycle. The gender difference obtained in our study may indicate an involvement of gonadal hormones in the control of the hydrolysis of aspirin. This possibility is currently under investigation.
Subject(s)
Female , Animals , Rats , Aspirin/pharmacokinetics , Carboxylic Ester Hydrolases/metabolism , Sex Differentiation , Adrenal Glands/enzymology , Brain/enzymology , Carboxylic Ester Hydrolases/blood , Estrus , Kidney/enzymology , Liver/enzymology , Rats, Wistar , Salicylates/blood , Sex FactorsABSTRACT
Adrenocortical autoantibodies (ACA), present in 60-80 percent of patients with idiopathic Addison's disease, are conventionally detected by indirect immunofluorescence (IIF) on frozen sections of adrenal glands. The large-scale use of IIF is limited in part by the need for a fluorescence microscope and the fact that histological sections cannot be stored for long periods of time. To circumvent these restrictions we developed a novel peroxidase-labelled protein A (PLPA) technique for the detection of ACA in patients with Addison's disease and compared the results with those obtained with the classical IIF assay. We studied serum samples from 90 healthy control subjects and 22 patients with Addison's disease, who had been clinically classified into two groups: idiopathic (N = 13) and granulomatous (N = 9). ACA-PLPA were detected in 10/22 (45 percent) patients: 9/13 (69 percent) with the idiopathic form and 1/9 (11 percent) with the granulomatous form, whereas ACA-IIF were detected in 11/22 patients (50 percent): 10/13 (77 percent) with the idiopathic form and 1/9 (11 percent) with the granulomatous form. Twelve of the 13 idiopathic addisonians (92 percent) were positive for either ACA-PLPA or ACA-IIF, but only 7 were positive by both methods. In contrast, none of 90 healthy subjects was found to be positive for ACA. Thus, our study shows that the PLPA-based technique is useful, has technical advantages over the IIF method (by not requiring the use of a fluorescence microscope and by permitting section storage for long periods of time). However, since it is only 60 percent concordant with the ACA-IIF method, it should be considered complementary instead of an alternative method to IIF for the detection of ACA in human sera
Subject(s)
Humans , Female , Aged , Middle Aged , Adult , Addison Disease/immunology , Adrenal Glands/enzymology , Autoantibodies/blood , Autoimmune Diseases/immunology , Immunoenzyme Techniques , Staphylococcal Protein A/immunology , Addison Disease/diagnosis , Aged, 80 and over , Fluorescent Antibody Technique, IndirectABSTRACT
La citocromo-c-oxidasa fue medida en adrenales de ratas neonatales de 1 a 10 días de edad y en adrenales de ratas adultas. No se encontraron diferencias entre las adrenales de machos y hembras neonatales de la misma edad. Por el contrario, las adultas mostraron diferencias sexuales. La actividad enzimática fue más elevada en los machos que en las hembras. En estas últimas se observaron cambios importantes durante el ciclo estral. Los valores más elevados se observaron durante el diestro y los más bajos durante el estro. Estos resultados proveen nuevas evidencias de la interrelación entre las glándulas gonadales y adrenales durante la maduración sexual
Subject(s)
Adrenal Glands/enzymology , Electron Transport Complex IV/metabolism , Adrenal Glands/physiology , Age Factors , Analysis of Variance , Estrus , Gonads/physiology , Sex Factors , Sexual MaturationABSTRACT
La 5 ß reductasa (Rasa) aumenta después de la eclosión en tejidos esteroidogénicos de la gallina en desarrollo, especialmente en el ovario derecho. La sintetasa del ácido delta aminolevulínico (ALAs) es más activa en estos tejidos que en hígado durante la mayoría de los estadios embrionarios. Pero después de la eclosión sólo aumenta en forma aguda ALAs hepática y adrenal; en ovario izquierdo dicha enzima crece moderadamente y en ovario derecho desciende a valores muy bajos. Cierta relación entre las curvas de ALAs y Rasa durante el desarrollo embrionario del ovario izquierdo y la adrenal sugieren que la 5 ß pregnanediona fuera un inductor natural de la ALAs en estas glándulas endocrinas funcionantes, por lo menos durante sus estadios embrionarios