Colestasis por deficiencia de 3β-Δ5-C27-hidroxiesteroide deshidrogenasa en un paciente con alteración en la síntesis de ácidos biliares / Cholestasis due to deficiency of 3β-Δ5-C27-hydroxysteroid dehydrogenase in a patient with altered bile acid synthesis

Resumen: Introducción: Los errores innatos en la síntesis de ácidos biliares son un grupo de defectos genéticos que representan del 1 al 2% de las enfermedades colestásicas crónicas en lactantes, niños y adolescentes. La deficiencia de 3β-Δ5-C27-hidroxiesteroide oxidoreductasa (3β-HSDH) es el defecto más comúnmente reportado. El cuadro clínico característico consiste en hepatitis neonatal, hepatomegalia, esplenomegalia, malabsorción, desnutrición y enfermedad hepática de aparición tardía. Caso clínico: Lactante masculino con antecedente de ictericia en escleras a los 4 meses que se resolvió espontáneamente; posteriormente, a los 18 meses, presentó enfermedad colestásica. Durante su abordaje se documentó gamma-glutamil transpeptidasa normal, hallazgo que es altamente sugestivo de alteración en la síntesis de ácidos biliares. El diagnóstico se realizó con espectrometría de masas en orina. Se inició tratamiento con ácido cólico oral, y presentó mejoría inmediata. Conclusiones: El resultado en los ácidos biliares urinarios es definitivo para el defecto genético y consistente con mutaciones homocigotas en el gen HSD3B7. Este padecimiento constituye un diagnóstico de exclusión en las enfermedades colestásicas de la infancia, particularmente el hallazgo de gamma-glutamil transpeptidasa normal o levemente aumentada, y responde adecuadamente al tratamiento oral, por lo que debe identificarse de forma temprana.

Abstract: Background: Inborn errors in bile acid synthesis are a group of genetic defects accounting for 1 to 2% of chronic cholestatic diseases in infants, children and adolescents. Deficiency of 3β-Δ5-C27-hydroxysteroid dehydrogenase (3β-HSDH) is the most common defect in this disease. Clinical features consist of neonatal hepatitis, hepatomegaly, splenomegaly, malabsorption, malnutrition, and late-onset liver disease. Case report: A male infant who presented jaundice in sclera at 4 months that resolved spontaneously, later presented cholestatic disease at 18 months. During his approach, normal gamma-glutamyl transpeptidase was documented, a finding that is highly suggestive of alteration in the synthesis of bile acids. The diagnosis was made using urine mass spectrometry. Oral colic acid treatment was started, presenting immediate improvement. Conclusions: The result in urinary bile acids is definitive for the genetic defect and consistent with homozygous mutations in the HSD3B7 gene. This condition is a diagnosis of exclusion in childhood cholestatic diseases, particularly in the presence of normal or mildly enlarged gamma-glutamyl transpeptidase, and responds adequately to oral treatment; it should be identified early.

Expression of steroidogenic enzymes in the rat model of polycystic ovary syndrome / 生理学报

The aim of the present study was to investigate the expression changes of three steroidogenic enzymes in the polycystic ovary syndrome (PCOS). Thirty Sprague-Dawley (SD) rats were randomly divided into normal control (NC) group and PCOS group. PCOS rat model was established by DHEA injection. The serum levels of progesterone, estrogen and testosterone were measured by immunoradioassay or enzyme immunoassay. The cellular distributions of 3β-hydroxy steroid dehydrogenase (3β-HSD), 17β-hydroxy steroid dehydrogenase (17β-HSD) and cytochrome P450 aromatase (P450arom) in ovaries were detected by immunohistochemistry. The expression levels of 3β-HSD, 17β-HSD and P450arom were detected by RT-PCR and Western blot. The results showed that the serum levels of estrogen and testosterone of PCOS group were significantly higher than those of the NC group. There was no significant difference of serum progesterone level between the PCOS and NC groups. Compared with the NC group, the PCOS group showed increased mRNA and protein expressions of both 3β-HSD and 17β-HSD, as well as reduced P450arom mRNA and protein expressions. These results suggest that 3β-HSD and 17β-HSD, but not P450arom, may participate in the ovarian hormonal regulation in the present rat model of PCOS.

Clinical features of a Chinese infant with inborn error of bile acid metabolism-3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency and review of the literature / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features of children with 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency and review the literature.</p><p><b>METHOD</b>Clinical features and treatment of one Chinese infant with 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency confirmed by HSD3B7 gene mutation analysis were retrospectively reviewed, and 51 cases of 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency who were internationally reported since 2000 were also reviewed in this paper.</p><p><b>RESULT</b>(1) A 3-month-old infant with neonatal cholestasis was admitted to our hospital because of hyperbilirubinemia and abnormal liver dysfunction (total bilirubin 110.7 µmol/L, direct bilirubin 74.5 µmol/L, γ-glutamyltransferase 24.4 IU/L, total bile acid 0.1 µmol/L).His jaundice disappeared within a few weeks, serum liver biochemistries improved and his growth in weight and height was excellent after oral cholic acid therapy.HSD3B7 gene analysis using peripheral lymphocyte genomic DNA from the patient identified compound heterozygous mutations. This child was confirmed as the most common inborn error of bile acid metabolism-3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency by molecular analysis.(2) Retrospective review of the literature showed that the clinical features of 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency included neonatal cholestasis, some patients progressed to severe liver disease and needed liver transplantation without effective therapy; however, serum biochemical characteristics of normal γ-glutamyltransferase activity, normal or low total bile acid concentrations were not consistent with cholestasis, the replacement treatment with cholic acid produced a dramatic improvements in symptoms, biochemical markers of liver injury; 31 cases were diagnosed by HSD3B7 gene mutation analysis.</p><p><b>CONCLUSION</b>The clinical characteristics of 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency include neonatal cholestasis, normal serum γ-glutamyltransferase activity, and normal or low serum total bile acid concentration.Oral cholic acid replacement is an effective therapy; definitive diagnosis of 3β-hydroxy-Δ(5)-C27-steroid dehydrogenase deficiency can be identified by molecular genetic testing technology.</p>

Primary culture, identification and functional study of rat Leydig cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To set up a stable primary culture system of Leydig cells with higher purity.</p><p><b>METHODS</b>We separated Leydig cells from other testicular cells, such as Sertoli and germ cells, by enzymatic digestion in combination with Percoll density gradient centrifugation and identified Leydig cells by 3beta-HSD staining.</p><p><b>RESULTS</b>The purity achieved by this method was above 95% and the total number of Leydig cells obtained from one testicle was about 1 x 10(6). The cytoplasm of Leydig cells was stained in deep blue by 3beta-HSD staining, and these cells possessed testosterone-secreting capability.</p><p><b>CONCLUSION</b>Leydig cells can be separated by enzymatic digestion combined with Percoll density gradient centrifugation, and 3beta-HSD staining to identify Leydig cells is simple and feasible with high purity.</p>

Gonadotrophin-releasing hormone-I and -II stimulate steroidogenesis in prepubertal murine Leydig cells in vitro / 亚洲男科学杂志(英文版)

<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>

Effect of exogenous selenium on the testicular toxicity induced by ethanol in rats.

The effects of supplementation of selenium at a dose of 10 microg/ kg body weight were investigated on ethanol induced testicular toxicity in rats. In the present study, four groups of male albino rats were maintained for 60 days, as follows: (1) Control group (normal diet) (2) Ethanol group (4g/kg body weight) (3) Selenium (10 microg/kg body weight) (4) Ethanol + Selenium (4g/kg body weight + 10 microg/kg body weight). Results revealed that ethanol intake caused drastic changes in the sperm count, sperm motility and sperm morphology. It also reduced the levels of testosterone and fructose. The activities of 3betaHSD, 17betaHSD in the testis and SDH in the seminal plasma were also reduced. Lipid peroxidation was also enhanced as the lipid peroxidation products were increased and the activities of the scavenging enzymes were reduced. But on coadministration of selenium along with alcohol all the biochemical parameters were altered to near normal levels indicating a protective effect of selenium. These results were reinforced by the histopathological studies.

Di(2-ethylhexyl) phthalate affects the testes and leydig cells of neonatal KM mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of di(2-ethylhexyl)phthalate (DEHP) on neonatal mice's testes and Leydig cells in vivo.</p><p><b>METHODS</b>Pregnant mice were exposed to DEHP at the dose of 100 mg/kg, 200 mg/kg or 500 mg/kg (body weight) per day by gavage from gestation day 12 (GD 12) through postnatal day 3 (PND 3), respectively. The testis and body weights, testicular histopathology and the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) of the neonatal mice were investigated.</p><p><b>RESULTS</b>The body and testis weights of the male mice's offspring were significantly reduced following DEHP exposure. Leydig cell morphology was affected significantly by DEHP as compared with the controls. Leydig cells obviously increased in the neonatal mice's testes on PND 15 and PND 30 when exposed to DEHP (500 mg/[kg x d]). Activities and positive area of the steroidogenic enzymes 3beta-HSD immunoexpression decreased markedly when exposed to DEHP (100 mg/[kg x d] or 200 mg/[kg x d]). Image analysis showed a decrease in the activities of 3beta-HSD in the animals exposed to DEHP (500 mg/[kg x d]), but an increase in the positive area of 3beta-HSD immunoexpression as compared with the control animals on PND 15 (P < 0.01).</p><p><b>CONCLUSION</b>DEHP affects the Leydig cell morphology, the activity of 3beta-HSD, the testis and body weights and the testicular histopathology of neonatal mice, and it may function as an antiandrogenic agent.</p>

The culture and identification of rat testis Leydig cell / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish a primary culture method of rat testis Leydig cell.</p><p><b>METHODS</b>The primary rat Leydig cells were treated with or without 4 U/ml human chorionic gonadotropin (hCG), and testosterone in culture medium was detected by radioimmunoassay. The morphology and biological characteristics of Leydig cell were observed.</p><p><b>RESULTS</b>The culture cells were highly homogeneous, proliferative and had a high differentiation rate. The high purified Leydig cells were verified by their dynamic morphological changes and 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD) histochemical staining. The testosterone secretion induced by hCG significantly increased (P < 0.05) 24 hours after inoculation than that induced without hCG in the control.</p><p><b>CONCLUSION</b>It suggests that the Leydig cell cultured in vitro may secrete high concentration of testosterone, and this study laid the basis of androgen replacement therapy for partial androgen deficiency in aging male.</p>

Effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity in rats.

In order to find out the effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity, male rats fed with ethanol 1.6g/kg body weight per day for four weeks were studied. Besides a drastic reduction in body and testis weight, there was decrease in ascorbic acid, reduced glutathione and activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase in the testicular tissue of the treated animals. Simultaneously, there was increase in lipid peroxidation and glutathione S-transferase activity. Activities of 3 beta-hydroxy steroid dehydrogenase and 17 beta-hydroxy steroid dehydrogenase were also found decreased in the treated animals. The results indicate that chronic ethanol administration resulted in increase in oxidative stress and decrease in the activities of steroidogenic enzymes in the rat testes.

Effects of fenvalerate on steroidogenesis in cultured rat granulosa cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This study was designed to examine the in vitro effects of fenvalerate on steroid production and steroidogenic enzymes mRNA expression level in rat granulosa cells.</p><p><b>METHODS</b>Using primary cultured rat granulosa cells (rGCs) as model, fenvalerate of various concentrations (0, 1, 5, 25, 125, 625 micromol/L) was added to the medium for 24 h. In some cases, optimal concentrations of 22(R)-hydroxycholesterol (25 micromol/L), Follicle stimulating hormone (FSH, 2 mg/L), or 8-Bromo-cAMP (1 mmol/L) were provided. Concentrations of 17 beta-estradiol(E2) and progesterone (P4) in the medium from the same culture wells were measured by RIA and the steroidogenic enzyme mRNA level was quantified by semi-quantitative RT-PCR.</p><p><b>RESULTS</b>Fenvalerate decreased both P4 and E2 production in a dose-dependent manner while it could significantly stimulate rGCs proliferation. This inhibition was stronger in the presence of FSH. Furthermore, it could not be reversed by 22(R)-hydroxycholesterol or 8-Bromo-cAMP. RT-PCR revealed that fenvalerate had no significant effect on 3 beta-HSD, but could increase the P450scc mRNA level. In addition, 17 beta-HSD mRNA level was dramatically reduced with the increase of fenvalerate dose after 24 h treatment.</p><p><b>CONCLUSION</b>Fenvalerate inhibits both P4 and E2 production in rGCs. These results support the view that fenvalerate is considered as a kind of endocrine-disrupting chemicals. The mechanism of its disruption may involve the effects on steroidogenesis signaling cascades and/or steroidogenic enzyme's activity.</p>

Effect of L-tyrosine on 3beta-HSD activity of rat luteal cells in vitro / 中国应用生理学杂志

<p><b>AIM</b>To study the effects of L-tyrosine on 3beta-HSD activity of rat luteal cells in vitro.</p><p><b>METHODS</b>Luteal cells were isolated from ovary tissues of female rats pretreated with PMSG and hCG. Luteal cells were cultured with 95% oxygen and 5% carbon dioxide in 37 degrees C. 3beta-HSD activity was measured by radioimmunoassay (RIA).</p><p><b>RESULTS</b>(1) 0.2 mmol x L(-1) and 2.0 mmol x L(-1) L-tyrosine significantly inhibited 3beta-HSD activity. (2) 0.2 mmol x L(-1) L-tyrosine exerted different effects on 3beta-HSD activity at different concentrations of pregnenolone (Ph). It increased 3beta-HSD activity at 0.1 micromol x L(-1) and 1 micromol x L(-1) of Pn concentration. With further increase in the concentration of Pn to 100 micromol x L(-1), the stimulating effect of L-tyrosine was switched to suppression effect. (3) L-tyrosine and L-tyrosine hydrazide both inhibited 3beta-HSD activity induced by hCG.</p><p><b>CONCLUSION</b>L-tyrosine affects 3beta-HSD activity of rat luteal cells in vitro. L-tyrosine and tyrosine hydrazide inhibits hCG induced 3beta-HSD activity.</p>

Protection of adrenocortical activity by dietary casein in ether anaesthetized rats.

Adrenal delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta-HSD) activity and serum corticosterone level were significantly higher in rats fed with 5% casein or 4% albumin diets after 1 hr of ether anaesthetic stress as compared to the controls, 5% casein and 20% casein (equivalent to 4% albumin) respectively. Ether anaesthesia to 20% casein fed rats caused no change in adrenal delta5-3beta-HSD activity and serum corticosterone level when compared with controls fed 20% casein diet. The results suggest that high milk protein diet may prevent acute stress effects by protecting adrenocortical activity. The present investigation opens up a new area of management of stress.

Impact of feeding ethanolic extracts of Achyranthes aspera Linn. on reproductive functions in male rats.

Feeding 50% ethanolic extract of A. aspera to male rats resulted in reduced sperm counts, weight of epididymis, serum level of testosterone and testicular activity of 3beta-hydroxysteroid dehydrogenase, while motility of the sperm and activity of the HMG CoA reductase were not affected. Cholesterol level in the testis, incorporation of labelled acetate into cholesterol, 17-ketosteroids in urine and hepatic and fecal bile acids were increased. The results suggest that ethanolic extract of A. aspera caused reproductive toxicity in male rats and the action may be by suppressing the synthesis of androgen.

Bases moleculares da hiperplasia adrenal congênita / Molecular bases of congenital adrenal hyperplasia

Hiperplasia adrenal congênita (HAC) é uma doença autossômica recessiva decorrente da alteraçäo de enzimas que participam da síntese do cortisol. As manifestações podem ser causadas pela deficiência do cortisol e, em alguns casos, aldosterona e pelo acúmulo de precursores. O objetivo desta revisäo é apresentar os mecanismos moleculares dos principais defeitos enzimáticos envolvidos na etiopatogênese da HAC. A deficiência da 21-hidroxilase (210H) ocorre em 95 por cento dos casos de HAC. Existem dois genes que codificam o P450c21: um ativo, CYP21, e um pseudogene CYP27P. Ambos säo altamente homólogos (98 por cento), o que favorece o emparelhamento desigual dos cromossomos homólogos durante a meiose, levando a duplicações e/ou deleções ou conversões desses genes. Adicionalmente, foram também descritas mutações de ponto, muitas delas presentes no pseudogene sugerindo microconversões. Mutações no gene CYP 1181 causam HAC por deficiência da 11 Rhidroxilase, forma esta que corresponde a 5 por cento dos casos. Algumas mutações säo recorrentes, situando-se principalmente entre os exons 6-8 que representaria uma área hot-spot no gene CYP 1781. A deficiência de 17-hidroxilase é causada por mutações no gene CYP77, que codificam uma proteína alterada, levando a deficiência total ou parcial de 17hidroxilaçäo e 17,20-liase ou deficiência isolada de 17,20-Base. Finalmente, deficiência de 3B-HSD é causada por mutações no gene HSD3B2, que codifica a enzima 3B-HSD tipo II e estas mutações têm sido associadas tanto com a forma clássica como com a forma näo clássica da deficiência da 3B-HSD.

Hiperplasia suprarrenal congénita / Congenital adrenal hyperplasia

La esteroidogénesis suprarrenal es un proceso complejo y secuencial que involucra a una serie de enzimas, las cuales actuando en forma coordinada sobre el colesterol determinan la síntesis de glucocorticoides y mineralocorticoides. El término hiperplasia suprarrenal congénita ha sido tradicionalmente usado para denominar al conjunto de alteraciones en la esteroidogénesis suprarrenal que determinan un decremento en la biosíntesis de cortisol. Este hecho determina una elevación compensatoria de ACTH, la cual, al estimular la síntesis esteroidal, lleva a un aumento de la producción de esteroides localizados antes del bloqueo. El resultado final es una diversidad de cuadros clínicos determinados por el déficit de cortisol y hormonas distales al bloqueo y al exceso de hormonas y metabolitos proximales al bloqueo. Los cuadros más frecuentes de hiperplasia suprarrenal congénita son los déficit de 21 y 11-hidroxilasa, que serán revisados en conjunto con otros déficit enzimáticos de presentación menos frecuente

Prevention of adrenocortical hyperactivity by dietary casein in rats exposed to forced swimming stress.

Adrenal weight, adrenal hydroxysteroid dehydrogenase activity and serum corticosterone level were significantly higher in rats fed with 5% casein diet after 7 days of swimming stress (45 min/day) as compared to their controls. All the parameters were similar to their control levels in rats receiving 20% casein diet and exposed to swimming stress. The results suggest that casein can play an important role in preventing adrenocortical hyperactivity in swimming stressed rats.

Effect of mercuric chloride on testicular activities in mice, Musculus albinus.

Impaired testicular function was observed after an exposure of Swiss albino mice (30 +/- 2 g) to mercuric chloride. A sublethal chronic exposure (0.5 ppm for 21 days) resulted in regressed histological and histochemical properties of the testis. The changes observed were degenerated tunica albuginea, abnormal configurations of seminiferous tubules, deformed primary and secondary spermatocytes, hypertrophy and vacuolization in interstitial cells and Sertoli cells. The 3 beta and 17 beta hydroxy steroid dehydrogenase enzyme and the level of testosterone hormone were significantly (p < 0.001) reduced. The diameter of different spermatogenic cells were significantly (p < 0.001) reduced.

Role of light in the mediation of acute effects of a single afternoon melatonin injection on steroidogenic activity of testis in the rat.

Young adult male rats, maintained either in an LD 12 : 12 or in continuous illumination (LL) for one week, were given a single injection of 25 microg melatonin/100 g body wt or ethanolic-saline (control) at 17.00 h. Animals from each group were sacrificed at 11.00 h on the following day. The activity of two important steroidogenic enzymes, 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) and delta(5)-3 beta-hydroxysteroid dehydrogenase (delta(5)-3 beta-HSD), and serum concentrations of testosterone, were measured following highly specific and sensitive spectrophotometric techniques and RIA, respectively. A significant decrease in the activity of both the steroidogenic enzymes was noted in the testes of melatonin-treated rats maintained under normal light-dark schedules, but this response was found to be lacking in the LL rats. However, no significant changes in the level of serum testosterone were noted in either group of melatonin-treated rats from the values in respective groups of ethanolic saline-administered LD and LL rats. Exposure of ethanolic saline-injected rats to continuous light also did not cause any change in the steroidogenic activity of the testis from those in LD rats. The study indicates that continuous light as such does not affect the endocrine function of testis but abolishes suppressive effects of melatonin on the steroidogenic activity of the testis in rat.

Gentamicin induced inhibition of steroidogenic enzymes in rat testis.

Gentamicin is an aminoglycoside antibiotic, widely used for treating many gram negative bacterial infections. Though nephrotoxicity is the most highlighted side effect, it has also been found to cause an alteration in the phosphatase activities of testes and accessory sex organs and a decline in the sperm count. This study was designed to assess the effects of gentamicin on testicular steroidogenesis and to ascertain whether such alterations are reversible. Laboratory inbred adult, male, 'Wistar' strain rats were chosen as the experimental animal. A significant dose-dependant reduction in the activities of the two steroidogenic enzymes, accompanied with a significant decrease in ascorbic acid and elevation of level of cholesterol was observed. The effects were maximum at a dose of 100 mg/kg, b.wt. After 15 days of withdrawal of the drug therapy the biochemical parameters namely ascorbic acid and cholesterol returned to normal levels whereas the activities of the two dehydrogenases showed a compensatory increase. This indicates that gentamicin affects the steroidogenic enzymes, causing an alteration in the formation of testosterone, which was manifested in the elevated cholesterol in the adult rat testes. However, these alterations were reversible.