RESUMEN
The potential inhibitory effects of flavonoids on gonadal steroid biosynthesis have gained attention due to their widespread presence in natural plant sources. Specifically, our study focused on evaluating the inhibitory efficacy of these compounds on human 3ß-hydroxysteroid dehydrogenase 2 (h3ß-HSD2) and rat homolog r3ß-HSD1, enzymes responsible for the conversion of pregnenolone to progesterone. Through our investigations, we observed that the potency of flavonoids was silymarin (IC50, 1.31 µM) > luteolin (4.63 µM) > tectorigenin > (5.86 µM), and rutin (44.12 µM) in inhibiting human KGN cell microsomal h3ß-HSD2. Similarly, the potency of flavonoids was silymarin (9.50 µM) > luteolin (11.49 µM) > tectorigenin (14.06 µM), and rutin (145.71 µM) in inhibiting rat testicular r3ß-HSD1. Silymarin, luteolin, and tectorigenin acted as mixed inhibitors of both human and rat 3ß-HSDs. Luteolin and tectorigenin were able to penetrate human KGN cells to inhibit progesterone secretion. Furthermore, docking analysis and structure-activity relationship analysis highlighted the importance of hydrogen bond formation for the inhibitory efficacy of these compounds against h3ß-HSD2 and r3ß-HSD1. Overall, this study demonstrates that silymarin exhibits the most potent inhibition of human and rat gonadal 3ß-HSDs, and significant SAR differences exist among the tested compounds.
Asunto(s)
Flavonoides , Silimarina , Humanos , Ratas , Animales , Flavonoides/farmacología , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Progesterona , Luteolina/farmacología , Relación Estructura-Actividad , Rutina/farmacología , 11-beta-Hidroxiesteroide DeshidrogenasasRESUMEN
As a functional fatty acid, α-linolenic acid (ALA) is essential in promoting animal testosterone biosynthesis. This study investigated the effects of ALA on testosterone biosynthesis and the possible mechanism underlying the signaling pathway in primary Leydig cells of the rooster. METHODS: Primary rooster Leydig cells were treated with ALA (0, 20, 40, or 80 µmol/L) or pretreated with a p38 inhibitor (50 µmol/L), a c-Jun NH2-terminal kinase (JNK) inhibitor (20 µmol/L), or an extracellular signal-regulated kinase (ERK) inhibitor (20 µmol/L) before ALA treatment. Testosterone content in the conditioned culture medium was detected using an enzyme-linked immunosorbent assay (ELISA). The expression of steroidogenic enzymes and JNK-SF-1 signaling pathway factors was detected using real-time fluorescence quantitative PCR (qRT-PCR). RESULTS: Supplementation with ALA significantly increased testosterone secretion within culture media (P < 0.05), and the optimized dose was 40 µmol/L. Compared with the control group, steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) mRNA expression significantly increased (P < 0.05) in the 40 µmol/L ALA group; 17-hydroxylase/c17-20 lyase (P450c17) and p38 mRNA expressions were not significantly different in the 40 µmol/L ALA group; ERK and JNK mRNA expressions were significantly upregulated (P < 0.05) in 40 µmol/L ALA group. In the inhibitor group, testosterone levels were significantly downregulated (P < 0.05). Compared with the 40 µmol/L ALA group, StAR, P450scc, and P450c17 mRNA expressions were significantly decreased (P < 0.05), and 3ß-HSD mRNA expression in the p38 inhibitor group did not change; StAR, P450scc, and 3ß-HSD mRNA expressions were significantly decreased (P < 0.05), and P450c17 mRNA expression in ERK inhibitor group did not change; StAR, P450scc, 3ß-HSD, and P450c17 mRNA expressions were significantly decreased (P < 0.05) in JNK inhibitor group. Additionally, the increased steroidogenic factor 1 (SF-1) gene expression levels induced by ALA were reversed when the cells were pre-incubated with JNK and ERK inhibitors. The levels in the JNK inhibitor group were significantly lower than those in the control group (P < 0.05). CONCLUSION: ALA may promote testosterone biosynthesis by activating the JNK-SF-1 signaling pathway to upregulate StAR, P450scc, 3ß-HSD, and P450c17 expression in primary rooster Leydig cells.
Asunto(s)
Células Intersticiales del Testículo , Ácido alfa-Linolénico , Masculino , Animales , Células Intersticiales del Testículo/metabolismo , Factor Esteroidogénico 1/metabolismo , Factor Esteroidogénico 1/farmacología , Ácido alfa-Linolénico/farmacología , Pollos/genética , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , ARN Mensajero/metabolismo , Testosterona/metabolismo , Transducción de Señal , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Chinese medicine, licorice (the roots of Glycyrrhiza glabra and G. inflata) has been used to treat inflammation and sexual debility for over 1000 years. Pharmacological studies have identified many biologically active chalcone derivatives from licorice. AIM OF THE STUDY: Human 3ß-Hydroxysteroid dehydrogenase 2 (h3ß-HSD2) catalyzes the formation of precursors for sex hormones and corticosteroids, which play critical roles in reproduction and metabolism. We explored inhibition and mode action of chalcones of inhibiting h3ß-HSD2 and compared it with rat 3ß-HSD1. MATERIALS AND METHODS: We investigated the inhibition of 5 chalcones on h3ß-HSD2 and compared species-dependent difference with 3ß-HSD1. RESULTS: The inhibitory strength on h3ß-HSD2 was isoliquiritigenin (IC50, 0.391 µM) > licochalcone A (0.494 µM) > licochalcone B (1.485 µM) > echinatin (1.746 µM) >chalcone (100.3 µM). The inhibitory strength on r3ß-HSD1 was isoliquiritigenin (IC50, 0.829 µM) > licochalcone A (1.165 µM) > licochalcone B (1.866 µM) > echinatin (2.593 µM) > chalcone (101.2 µM). Docking showed that all chemicals bind steroid and/or NAD+-binding site with the mixed mode. Structure-activity relationship analysis showed that strength was correlated with chemical's hydrogen bond acceptor. CONCLUSION: Some chalcones are potent h3ß-HSD2 and r3ß-HSD1 inhibitors, possibly being potential drugs to treat Cushing's syndrome or polycystic ovarian syndrome.
Asunto(s)
Chalcona , Chalconas , Glycyrrhiza , Humanos , Ratas , Animales , Chalconas/farmacología , Chalcona/farmacología , Glycyrrhiza/química , Hidroxiesteroide Deshidrogenasas , 3-Hidroxiesteroide Deshidrogenasas/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Chinese medicine, curcuma longa L has been applied to treat pain and tumour-related symptoms for over thousands of years. Curcuminoids, polyphenolic compounds, are the main pharmacological component from the rhizome of Curcuma longa L. Pharmacological investigations have found that curcuminoids have many pharmacological activities of anti-inflammatory, anti-tumour, and anti-metastasis. AIM OF THE STUDY: 3ß-Hydroxysteroid dehydrogenase (3ß-HSD1) catalyses the production of steroid precursors for androgens and estrogens, which play an essential role in cancer metastasis. We explored the potency and mode of action of curcuminoids and their metabolites of inhibiting 3ß-HSD1 activity and compared the species difference between human and rat. MATERIALS AND METHODS: In this study, we investigated the direct inhibition of 6 curcuminoids on human placental 3ß-HSD1 activity and compared the species-dependent difference in human 3ß-HSD1 and rat placental homolog 3ß-HSD4. RESULTS: The inhibitory potency of curcuminoids on human 3ß-HSD1 was demethoxycurcumin (IC50, 0.18 µM) > bisdemethoxycurcumin (0.21 µM)>curcumin (2.41 µM)> dihydrocurcumin (4.13 µM)>tetrahydrocurcumin (15.78 µM)>octahydrocurcumin (ineffective at 100 µM). The inhibitory potency of curcuminoids on rat 3ß-HSD4 was bisdemethoxycurcumin (3.34 µM)>dihydrocurcumin (5.12 µM)>tetrahydrocurcumin (41.82 µM)>demethoxycurcumin (88.10 µM)>curcumin (137.06 µM)> octahydrocurcumin (ineffective at 100 µM). Human choriocarcinoma JAr cells with curcuminoid treatment showed that these chemicals had similar potency to inhibit progesterone secretion under basal and 8bromo-cAMP stimulated conditions. Docking analysis showed that all chemicals bind pregnenolone-binding site with mixed/competitive mode for 3ß-HSD. CONCLUSION: Some curcuminoids are potent human placental 3ß-HSD1 inhibitors, possibly being potential drugs to treat prostate cancer and breast cancer.
Asunto(s)
Curcumina , Animales , Femenino , Humanos , Embarazo , Ratas , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Curcuma/química , Curcumina/química , Diarilheptanoides/farmacología , Hidroxiesteroide Deshidrogenasas/metabolismo , Placenta/metabolismo , Relación Estructura-ActividadRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Roots of Argyreia nervosa (Burm.f.) Bojer is used traditionally as an aphrodisiac and mentioned in the indigenous system of medicine as spermatogenic. The roots of the plant are also used as bitter, tonic, and alternative. AIM OF THE STUDY: To study the effect of n-butanol fraction (BTF) and ethyl acetate fraction (ETF) of methanol extract prepared from the roots of Argyreia nervosa and scopoletin isolated from ETF on testosterone biosynthesis in testis and spermatogenesis using rats. MATERIALS AND METHODS: The effect of BTF, ETF, and scopoletin on the testosterone biosynthesis was evaluated by determining the alteration in expression of mRNA corresponding to steroidogenic enzymes and concentration of testosterone using TM-3 cell line. The ability of BTF and ETF in altering the level of testicular cholesterol and testosterone along with mRNA expression corresponding to 3ß-Hydroxy-Δ5-steroid dehydrogenase (3ß-HSD) and Acute Steroid Regulatory Protein (StAR) was evaluated using rats as experimental animals. The sperm concentration in the seminal fluid was determined, and histological studies of testicular tissues were also carried out. RESULTS: Test solutions containing BTF, ETF, and scopoletin showed a dose-dependent and statistically significant increase in the testosterone content when incubated with TM-3 cells. The test solutions also increased the fold expression of mRNA corresponding to StAR and 3ß-HSD enzymes from TM-3 cells. BTF and ETF elevated testicular testosterone levels by 3.57 and 3.84-fold as compared to control animals, while the fractions showed 9.04 and 10.41-fold alteration in expression of mRNA corresponding to StAR, respectively. BTF and ETF altered the expression of mRNA corresponding to 3ß-HSD by 13.43 and 15.04-fold in testicular tissues; moreover, they elevated the activity of 3ß-HSD by 7.11 and 7.73 fold, respectively. The animals treated with BTF and ETF showed increased sperm concentration. Histological observations showed that the lumen of seminiferous tubules was densely populated with spermatozoa and Leydig cells were intensely stained. Extract prepared from fruits of Tribulus terrestris Linn and testosterone served as positive controls. CONCLUSION: BTF, ETF, and scopoletin could promote testosterone biosynthesis by elevating mRNA expression corresponding to StAR, 3ß-HSD, and by increasing 3ß-HSD activity in the testicular tissues. Elevated testosterone concentration in testis promoted spermatogenesis. The studies provided the probable mechanism through which the roots of A. nervosa act as spermatogenic.
Asunto(s)
Convolvulaceae/química , Extractos Vegetales/farmacología , Espermatogénesis/efectos de los fármacos , Testosterona/biosíntesis , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Células Intersticiales del Testículo/efectos de los fármacos , Células Intersticiales del Testículo/metabolismo , Masculino , Ratones , Extractos Vegetales/administración & dosificación , Raíces de Plantas , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Testículo/efectos de los fármacos , Testículo/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Quite a few plants are in use to treat female infertility and associated problems. Availing the cues from traditional knowledge, phytochemical studies and ethnopharmacological evidences, the aphrodisiac plant Ficus religiosa (F. religiosa) is widely in use to cure infertility in women. For instance, the juice of leaf and aerial root of F. religiosa is reported to normalize the dysregulated menstrual cycle in women. Besides, it is believed that regular circumambulation of F. religiosa during the early hours of the morning helps women in alleviating infertility which could be attributed to the potential phytovolatiles released from F. religiosa. However, the evidences for therapeutic potential of F. religiosa in treating female infertility are arbitrary and mostly anecdotal. AIM OF THE STUDY: The present study was aimed at examining if extracts of fresh and/or dry leaf of F. religiosa would cure polycystic ovary syndrome (PCOS) in the rat model. METHODS: Rats were divided into seven groups; control (Group I), PCOS-induced (P.O, Letrozole -1 mg/kg BW for 21 days) and untreated (Group II), PCOS-induced and treated with the leaf extracts of F. religiosa (Groups III-VI), and, PCOS-induced and treated with pioglitazone (Group VII). The estrous intervals, body and organ weights (ovary and uterus), and serum hormones (testosterone, luteinizing hormone [LH], estrogen, and progesterone) were measured, and the expression of Cyp19a1 (aromatase), and Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) were assessed in the experimental rats. The levels of 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and antioxidants (MDA, GSH, GPx, SOD, and CAT) were also quantified. Besides, the putative volatile compounds in the esterified leaf extracts were identified using Gas Chromatography-Mass Spectrometry (GC-MS). RESULTS: Letrozole treatment induced irregular estrous and altered weight of organs and hormonal milieu, which were reverted to normal in leaf extracts-treated PCOS-induced rats. Remarkably, fresh leaf treatment up-regulated Cyp19a1and PPAR-γ and increased the levels of 3ß-HSD and 17ß-HSD. We found 3-acetoxy-3-hydroxy-propionic acid in fresh and dry leaf extracts, which we attribute to efficacy of the extracts in alleviating PCOS. CONCLUSION: Put together, our findings suggest the leaves of F. religiosa as potential in alleviating PCOS, mainly due to the presence of putative volatile molecules. Further screening of the leaves of F. religiosa is recommended to identify other key molecules and to develop a systematic therapeutic intervention for PCOS.
Asunto(s)
Aromatasa/metabolismo , Ficus , Hormonas Esteroides Gonadales/biosíntesis , Ovario/efectos de los fármacos , PPAR gamma/metabolismo , Extractos Vegetales/farmacología , Síndrome del Ovario Poliquístico/tratamiento farmacológico , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Aromatasa/genética , Modelos Animales de Enfermedad , Femenino , Ficus/química , Ovario/enzimología , PPAR gamma/genética , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta , Síndrome del Ovario Poliquístico/enzimología , Síndrome del Ovario Poliquístico/genética , Ratas Wistar , Transducción de Señal , Regulación hacia ArribaRESUMEN
In a healthy female reproductive system, a subtle hormonal and metabolic dance leads to repetitive cyclic changes in the ovaries and uterus, which make an effective ovulation and potential implantation of an embryo possible. However, that is not so in the case of polycystic ovary syndrome (PCOS), in which case the central mechanism responsible for entraining hormonal and metabolic rhythms during the menstrual cycle is notably disrupted. In this review we provide a detailed description of the possible scenario of PCOS pathogenesis. We begin from the analysis of how a set of genetic disorders related to PCOS leads to particular malfunctions at a molecular level (e.g., increased enzyme activities of cytochrome P450 (CYP) type 17A1 (17α-hydroxylase), 3ß-HSD type II and CYP type 11A1 (side-chain cleavage enzyme) in theca cells, or changes in the expression of aquaporins in granulosa cells) and discuss further cellular- and tissue-level consequences (e.g., anovulation, elevated levels of the advanced glycation end products in ovaries), which in turn lead to the observed subsequent systemic symptoms. Since gene-editing therapy is currently out of reach, herein special emphasis is placed on discussing what kinds of drug targets and which potentially active substances seem promising for an effective medication, acting on the primary causes of PCOS on a molecular level.
Asunto(s)
Hormonas/metabolismo , Síndrome del Ovario Poliquístico , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Acuaporinas/metabolismo , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Femenino , Células de la Granulosa/enzimología , Células de la Granulosa/patología , Humanos , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Síndrome del Ovario Poliquístico/enzimología , Síndrome del Ovario Poliquístico/genética , Esteroide 17-alfa-Hidroxilasa/metabolismo , Células Tecales/enzimología , Células Tecales/patologíaRESUMEN
The present study aimed to determine whether an i.c.v. administration of allopregnanolone (ALLO) rapidly modifies the hypothalamic and ovarian 3ß-hydroxysteroid dehydrogenase (3ß-HSD) enzymatic activity and gene expression in in vivo and ex vivo systems in pro-oestrus (PE) and dioestrus I (DI) rats. Animals were injected with vehicle, ALLO, bicuculline or bicuculline plus ALLO and were then killed. In the in vivo experiment, the hypothalamus, ovaries and serum were extracted and analysed. In the ex vivo experiment, the superior mesenteric ganglion - ovarian nerve plexus - ovary system was extracted and incubated during 120 minutes at 37 ºC. The serum and ovarian compartment fluids were used to determine progesterone by radioimmunoanalysis. In the in vivo experiments, ALLO caused a decrease in hypothalamic and ovarian 3ß-HSD enzymatic activity during PE. During DI, ALLO increased hypothalamic and ovarian 3ß-HSD activity and gene expression. The ovarian 3ß-HSD activity increased in both stages in the ex vivo system; gene expression increased only during DI. ALLO induced an increase in serum progesterone only in D1 and in the ovarian incubation liquids in both stages. All findings were reversed by an injection of bicuculline before ALLO. Ovarian steroidogenic changes could be attributed to signals coming from ganglion neurones, which are affected by the acute central neurosteroid stimulation. The i.c.v. administration of ALLO via the GABAergic system altered 3ß-HSD activity and gene expression, modulating the neuroendocrine axis. The present study reveals the action that ALLO exerts on the GABAA receptor in both the central and peripheral nervous system and its relationship with hormonal variations. ALLO is involved in the "fine tuning" of neurosecretory functions as a potent modulator of reproductive processes in female rats.
Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/metabolismo , Hipotálamo/efectos de los fármacos , Neuroesteroides/administración & dosificación , Ovario/efectos de los fármacos , Pregnanolona/administración & dosificación , Animales , Diestro/efectos de los fármacos , Diestro/metabolismo , Femenino , Expresión Génica/efectos de los fármacos , Hipotálamo/enzimología , Inyecciones Intraventriculares , Ovario/metabolismo , Proestro/efectos de los fármacos , Proestro/metabolismo , Progesterona/sangre , RatasRESUMEN
Incidents of male infertility are mushrooming worldwide. Oxidative stress plays a prime role for its onset. Considering this background, the study was designed to focus the direct role of lycopene on cyproterone acetate (CPA) induced testicular hypofunction in rat. Four groups have been considered including the vehicle-treated control, lycopene-treated control, CPA-treated and CPA+ lycopene-treated groups. Androgenic, antioxidant and toxicity profiles were assessed. Results focused a nonsignificant (p > .05) difference in recovery of testicular Δ5 , 3ß-hydroxysteroid dehydrogenase (HSD), 17ß-HSD after direct exposure of lycopene compared to the CPA-treated group. On other side, lycopene exposure to the testicular tissue of CPA-treated rat (CPA+ lycopene-treated) exhibited a significant (p < .05, p < .001) rectification in testicular catalase, superoxide dismutase, peroxidase, glutathione-S-transferase activities towards the vehicle- and lycopene-treated control groups. Toxicity profile also showed a significant (p < .001) recovery in CPA-treated group after direct exposure of lycopene towards the vehicle- and lycopene-treated control groups. So, it can be concluded that direct exposure of lycopene may rectify the CPA-induced testicular hypofunction either by its free radical-quenching ability or by stimulating antioxidant enzyme activity without modulating androgenic key enzyme directly.
Asunto(s)
Acetato de Ciproterona/toxicidad , Depuradores de Radicales Libres/farmacología , Infertilidad Masculina/prevención & control , Licopeno/farmacología , Testículo/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Andrógenos/metabolismo , Animales , Catalasa/metabolismo , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Depuradores de Radicales Libres/uso terapéutico , Glutatión Transferasa/metabolismo , Humanos , Infertilidad Masculina/inducido químicamente , Licopeno/uso terapéutico , Masculino , Estrés Oxidativo/efectos de los fármacos , Peroxidasa/metabolismo , Ratas , Ratas Wistar , Recuento de Espermatozoides , Superóxido Dismutasa/metabolismo , Testículo/enzimologíaRESUMEN
The cholesterol hydroxylase/lyase (CHL) system, consisting of cytochrome P450scc, adrenodoxin (Adx) and adrenodoxin reductase (AdR), initiates mammalian steroidogenesis, converting cholesterol to pregnenolone. The foot-and-mouth disease virus 2A-based method allows to express multiple proteins from a single transcript. We developed a 2A-based multicistronic system for the coexpression of three bovine CHL system proteins as the self-processing polyprotein pCoxIV-P450scc-2A-Adx-2A-AdR-GFP (pCoxIV-CHL-GFP), with a cleavable N-terminal mitochondrial targeting presequence. HEK293T cells transfected with plasmid, containing complementary DNA (cDNA) for pCoxIV-CHL-GFP, efficiently performed the expression of P450scc-2A, targeted to mitochondria, and Adx-2A, AdR-GFP and the fusion protein Adx-2A-AdR-GFP, which were predominantly localized in the cytosol. Despite the spatial separation of expressed P450scc and redox partners, the transfected HEK293T cells were able to convert the steroid substrates of cytochrome P450scc to pregnenolone, whereas control HEK293T cells were not catalytically active. The presence of 2Ð peptide residue on the C-terminus of P450scc did not preclude its enzymatic activity. HEK293T cells transfected with a vector directing the synthesis of only P450scc-2A demonstrated cytochrome P450scc activity comparable to that of cells expressing all three CHL system components, and to that of nature steroidogenic cells. Thus, the P450scc activity detected in cells transfected with both constructed plasmids was the result of the effective functional coupling of the bovine cytochrome P450scc and endogenous mitochondrial electron transport proteins of HEK293T cells. The produced pregnenolone did not undergo further conversion to progesterone, which indicates the absence of catalytically active 3ß-hydroxysteroid dehydrogenase. Therefore, HEK293T cells may be suitable for the expression of steroidogenic enzymes and the study of their characteristics.
Asunto(s)
Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Mitocondrias/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Adrenodoxina/metabolismo , Western Blotting , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Cromatografía Líquida de Alta Presión , Ferredoxina-NADP Reductasa , Citometría de Flujo , Células HEK293 , Humanos , Microscopía Fluorescente , Plásmidos/genética , Pregnenolona/metabolismoRESUMEN
Fish like higher animals, have a well-defined mechanism to produce sex steroids that play a critical role in gonadal development and maturation. In this study, we aimed to analyse the expression pattern of 3ß-HSD in different tissues, during ontogenetic development and gonadal recrudescence of Clarias batrachus. A full-length cDNA of 1617 bp including an open reading frame (ORF) of 1125 bp encoding 374 amino acids was isolated from testes of C. batrachus. The docking analysis between C. batrachus 3ß-HSD protein and eurycomanone exhibited high binding affinity toward each other with total energy of -108.292 kcal/mol and van der Waals (VDW) interaction of -84.2838 kcal/mol. The 3ß-HSD transcript level during ontogeny was detected in all the stages starting from the fertilized egg. The mature C. batrachus showed more expression of 3ß-HSD mRNA in gonads and brain while weak expression was detected in the remaining tissues analysed. The 3ß-HSD mRNA expression during annual reproductive phases of gonads was more in preparatory and pre-spawning stages than that of spawning and post-spawning phases. The mRNA expression results together suggest that 3ß-HSD plays an important role in gonadal development. Furthermore, the active binding sites on 3ß-HSD protein could be targeted in pharmacological drug designing to cope with reproductive dysfunctions in fish.
Asunto(s)
Bagres/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Regulación Enzimológica de la Expresión Génica/fisiología , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bagres/crecimiento & desarrollo , Clonación Molecular , Biología Computacional , Femenino , Masculino , Modelos Moleculares , Estructura Molecular , Filogenia , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Extractos Vegetales/farmacología , Unión Proteica , Conformación Proteica , Cuassinas/química , Cuassinas/metabolismo , Cuassinas/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reproducción/fisiologíaRESUMEN
The aim of this study was to investigate the beneficial effects of zinc (Zn) in preventing lead (Pb)-induced reproductive toxicity in Wistar rats. The rats were divided into four groups, namely, control group, Pb group, Zn group, and Pb + Zn group. Animals were exposed to Pb (819 mg of Pb/L) or Zn (71 mg of Zn/L) or both through drinking water for 65 days. Rats exposed to Pb showed decreased weights of testes and accessory sex organs. Significant decrease in the testicular daily sperm production, epididymal sperm count, motility, viability, and number of hypoosmotic tail coiled sperm was observed in Pb-exposed rats. Testicular 3ß- and 17ß-hydroxysteroid dehydrogenase activity levels and circulatory testosterone levels were also decreased significantly in Pb-exposed rats. A significant increase in the lipid peroxidation products with a significant decrease in the activities of catalase and superoxide dismutase were observed in the testes and epididymis of Pb-exposed rats. Moreover, the testicular architecture showed lumens devoid of sperm in Pb-exposed rats. Supplementation of Zn mitigated Pb-induced oxidative stress and restored the spermatogenesis and steroidogenesis in Pb-exposed rats. In conclusion, cotreatment of Zn is effective for recovering suppressed spermatogenesis, steroidogenesis, elevated oxidative status, and histological damage in the testis of rats treated with Pb.
Asunto(s)
Suplementos Dietéticos , Epidídimo/efectos de los fármacos , Infertilidad Masculina/prevención & control , Intoxicación por Plomo/prevención & control , Estrés Oxidativo/efectos de los fármacos , Testículo/efectos de los fármacos , Zinc/uso terapéutico , 17-Hidroxiesteroide Deshidrogenasas/antagonistas & inhibidores , 17-Hidroxiesteroide Deshidrogenasas/química , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/antagonistas & inhibidores , 3-Hidroxiesteroide Deshidrogenasas/química , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Suplementos Dietéticos/efectos adversos , Epidídimo/metabolismo , Epidídimo/patología , Infertilidad Masculina/etiología , Intoxicación por Plomo/metabolismo , Intoxicación por Plomo/patología , Intoxicación por Plomo/fisiopatología , Peroxidación de Lípido/efectos de los fármacos , Masculino , Tamaño de los Órganos/efectos de los fármacos , Compuestos Organometálicos/antagonistas & inhibidores , Compuestos Organometálicos/toxicidad , Sustancias Protectoras/efectos adversos , Sustancias Protectoras/uso terapéutico , Distribución Aleatoria , Ratas Wistar , Espermatogénesis/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Espermatozoides/patología , Testículo/metabolismo , Testículo/patología , Testosterona/sangre , Enfermedades Transmitidas por el Agua/metabolismo , Enfermedades Transmitidas por el Agua/patología , Enfermedades Transmitidas por el Agua/fisiopatología , Enfermedades Transmitidas por el Agua/prevención & control , Zinc/efectos adversosRESUMEN
CONTEXT: Soy is the main source of phytoestrogens, which has long been used as traditional food. One major subtype of phytoestrogens includes isoflavones and they are scientifically validated for their beneficial actions on many hormone-dependent conditions. OBJECTIVE: The present study examines the effect of soy isoflavones on letrozole-induced polycystic ovary syndrome (PCOS) rat model. MATERIALS AND METHODS: PCOS was induced in Sprague-Dawley rats with of 1 mg/kg letrozole, p.o. once daily for 21 consecutive days. Soy isoflavones (50 and 100 mg/kg) was administered for 14 days after PCOS induction. Physical parameters (body weight, oestrous cycle determination, ovary and uterus weight) metabolic parameters (oral glucose tolerance test, total cholesterol), steroidal hormone profile (testosterone and 17ß-oestradiol), steroidogenic enzymes (3ß-hydroxy steroid dehydrogenase (HSD) and 17ß-HSD), oxidative stress and histopathology of ovary were studied. RESULTS: Soy isoflavones (100 mg/kg) treatment significantly altered the letrozole-induced PCOS symptoms as observed by decreased body weight gain (p < 0.05), percentage diestrous phase (p < 0.001), testosterone (p < 0.001), 3ß-HSD (p < 0.01) and 17ß-HSD (p < 0.001) enzyme activity and oxidative stress. Histological results reveal that soy isoflavones treatment in PCOS rats resulted in well-developed antral follicles and normal granulosa cell layer in rat ovary. DISCUSSION: Treatment with soy isoflavones exerts beneficial effects in PCOS rats (with decreased aromatase activity) which might be due to their ability to decrease testosterone concentration in the peripheral blood. CONCLUSION: Analysis of physical, biochemical and histological evidences shows that soy isoflavones may be beneficial in PCOS.
Asunto(s)
Antagonistas de Andrógenos/farmacología , Glycine max/química , Isoflavonas/farmacología , Nitrilos , Ovario/efectos de los fármacos , Fitoestrógenos/farmacología , Síndrome del Ovario Poliquístico/prevención & control , Triazoles , Útero/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Antagonistas de Andrógenos/aislamiento & purificación , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Biomarcadores/sangre , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Colesterol/sangre , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Estradiol/sangre , Ciclo Estral/efectos de los fármacos , Femenino , Isoflavonas/aislamiento & purificación , Letrozol , Ovario/metabolismo , Ovario/patología , Estrés Oxidativo/efectos de los fármacos , Fitoestrógenos/aislamiento & purificación , Fitoterapia , Plantas Medicinales , Síndrome del Ovario Poliquístico/sangre , Síndrome del Ovario Poliquístico/inducido químicamente , Síndrome del Ovario Poliquístico/patología , Ratas Sprague-Dawley , Testosterona/sangre , Factores de Tiempo , Útero/metabolismo , Útero/patología , Aumento de Peso/efectos de los fármacosRESUMEN
The present study was undertaken to understand the physiological significance of the existence of nitric oxide synthase (NOS)/nitric oxide (NO) system in fish ovary. For this, two doses of NO donor, sodium nitroprusside (SNP, 25 µg and 50 µg) and NOS inhibitor, N-nitro-l-arginine methyl ester (l-NAME, 50 µg and 100 µg)/100 g body weight were administered during the two reproductive phases of reproductive cycle of the Clarias batrachus During the late-quiescence phase, high dose of l-NAME decreased the NO, testosterone, 17ß-estradiol, vitellogenin contents in serum and ovary and activities of 5-ene-3ß-hydroxysteroid dehydrogenases (3ß-HSD) and 17ß-hydroxysteroid dehydrogenases (17ß-HSD) in ovary, whereas higher dose of SNP increased these parameters. l-NAME also reduced oocytes-I but increased perinucleolar oocytes in the ovary, whereas SNP treatment increased the number of advanced oocytes (oocytes-I and II) than the perinucleolar oocytes when compared with control ovary. During the mid-recrudescence phase, both doses of SNP increased NO, testosterone, 17ß-estradiol and vitellogenin in serum and ovary; however, l-NAME treatment lowered their levels. The activities of ovarian 3ß-HSD and 17ß-HSD were also stimulated by SNP, but l-NAME suppressed their activities compared to the control. The SNP-treated ovaries were dominated by oocyte-II and III stages, whereas l-NAME-treated ovary revealed more perinucleolar oocytes and oocytes-I and practically no advanced oocytes. Expression of endothelial NOS (eNOS), inducible NOS (iNOS) and neuronal NOS (nNOS) was augmented by the SNP and declined by l-NAME treatments as compared to the control. This study, thus, provides distinct evidence of NO-stimulated steroidogenesis, vitellogenesis and folliculogenesis in fish.
Asunto(s)
Peces/fisiología , Óxido Nítrico/fisiología , Folículo Ovárico/crecimiento & desarrollo , Esteroides/biosíntesis , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Inhibidores Enzimáticos , Estradiol/análisis , Estradiol/sangre , Femenino , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/análisis , Óxido Nítrico/antagonistas & inhibidores , Donantes de Óxido Nítrico/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/fisiología , Nitroprusiato/farmacología , Oocitos/química , Oocitos/efectos de los fármacos , Oocitos/fisiología , Ovario/enzimología , Ovario/fisiología , Testosterona/análisis , Testosterona/sangre , Vitelogeninas/análisis , Vitelogeninas/sangreRESUMEN
Removing dietary phytoestrogens causes obesity and diabetes in adult male rats. Based on the facts that hypothalamic food intake control is disrupted in phytoestrogen-deprived animals and that several steroids affect food intake, we hypothesized that phytoestrogen withdrawal alters the expression of hypothalamic steroidogenic enzymes. Male Wistar rats fed with a high-phytoestrogen diet from conception to adulthood were subjected to phytoestrogen withdrawal by feeding them a low-phytoestrogen diet or a high-phytoestrogen, high-fat diet. Withdrawal of dietary phytoestrogens increased 3ß-hydroxysteroid dehydrogenase and P450 aromatase gene expression and decreased those of 5α-reductase-1. This is a direct effect of the lack of dietary phytoestrogens and not a consequence of obesity, as it was not observed in high-fat-fed rats. Phytoestrogen withdrawal and high-fat diet intake reduced hypothalamic expression of estrogen receptor (ER)α correlated with low levels of ERα-O, ERα-OS, and ERα-OT transcripts. Variations in gene expression of steroidogenic enzymes may affect the content of neurosteroids. As neurosteroids are related to food intake control, the changes observed may be a novel mechanism in the regulation of energy balance in obese phytoestrogen-deprived animals. In rats, steroidogenesis and ER signaling appear to be altered by phytoestrogen withdrawal in the rat. The ubiquity of phytoestrogens in the diet and changing intakes or withdrawal suggest that aspects of human health could be affected based on the rat and warrant further research.
Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/metabolismo , Regulación del Apetito , Aromatasa/metabolismo , Colestenona 5 alfa-Reductasa/metabolismo , Dieta , Obesidad/etiología , Fitoestrógenos/administración & dosificación , Animales , Dieta Alta en Grasa , Ingestión de Alimentos/fisiología , Ingestión de Energía , Receptor alfa de Estrógeno/metabolismo , Expresión Génica , Hipotálamo/metabolismo , Masculino , Neurotransmisores/metabolismo , Obesidad/metabolismo , Fitoestrógenos/farmacología , Ratas Wistar , Transducción de SeñalRESUMEN
Antifertility efficacy of oral administration of aqueous fruit extract of Mimusops elengi (200, 400 and 600 mg kg(-1) body weight/day for 35 days) was evaluated in Parkes strain male mice. Various reproductive end points such as histopathology, sperm parameters, testosterone level, haematology, serum biochemistry and fertility indices were assessed; activities of 3ß- and 17ß-hydroxysteroid dehydrogenases, and immunoblot expressions of StAR and P450scc in the testis were also assessed. Histologically, testes in Mimusops-treated mice showed nonuniform and diverse degenerative changes in the seminiferous tubules; both affected and normal tubules were observed in the same sections of testis. The treatment had adverse effects on testicular hydroxysteroid dehydrogenases and StAR and P450scc, serum level of testosterone and on motility, viability and number of spermatozoa in cauda epididymis. However, serum levels of alanine aminotransferase, aspartate aminotransferase and creatinine, and haematological parameters were not affected by the treatment. Also, libido was not affected in treated males, but their fertility was markedly suppressed. By 56 days of treatment withdrawal, the alterations caused in the above parameters recovered to control levels, suggesting that Mimusops treatment causes reversible suppression of spermatogenesis and fertility in Parkes mice. Further, there were no detectable signs of toxicity in treated males.
Asunto(s)
Fertilidad/efectos de los fármacos , Frutas/química , Mimusops/química , Extractos Vegetales/farmacología , Espermatogénesis/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Masculino , Ratones , Fosfoproteínas/metabolismo , Túbulos Seminíferos/patología , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Testículo/patología , Testosterona/sangre , Testosterona/metabolismoRESUMEN
BACKGROUND: Colorectal cancer (CRC) is the third most common cancer. Its global incidence and mortality have been on the rise. Recent strategy of therapies has involved the use of non-steroid anti-inflammatory drugs and cyclooxygenase-selective inhibitors. Aerial parts of Imperata cylindrical L. Raeusch (IMP) have been used as an anti-inflammatory agent in traditional Chinese medicine. HYPOTHESIS: Asarachidonate acid cascadeis often involved in inflammation-related malignancy and IMP is an anti-inflammatory agent, hence it is hypothesized that IMP aerial part ethyl acetate extract exerts cytotoxic effects on colorectal cancer cells in vitro. STUDY DESIGN: The HT-29 adenocarcinoma cell line was used to elucidate its pro-apoptotic activities. Flow cytometry and fluorescent microscopy were performed to assess cell cycle arrest and the accumulation of reactive oxygen species (ROS). The mRNA and hormone levels of arachidonate acid pathways were studied via quantitative reverse transcription PCR (qRT-PCR) and ELISA. RESULTS: The 50% growth inhibitory effect (GI50) of the IMP extract on HT-29 was measured with a value of 14.5 µg/ml. Immuno-blot and caspase-3/7 activity assay showed the pro-apoptotic effect of IMP on the activation of caspase cascade. G2/M arrest was observed via flow cytometry. The ROS activity was modulated by the IMP extraction a concentration-dependent manner in HT-29 cells. The IMP extract increased PGE2 and PGF2α levels qRT-PCR revealed that transcripts of rate-limiting PGE2- and PGF2α-biosynthetic enzymes - COX-1, mPGES1 and AKR1C3 were notably up-regulated. Among the prostanoid receptors, EP1 and FP transcripts were up-regulated while EP4 transcripts decreased. The findings suggest that the proliferative effect of PGE2, which is generally believed to associate with heightened DNA synthesis and cross-talk with MAPK pathways, is likely triggered by the pro-apoptotic or -oxidative effects exerted by IMP extract in HT-29 cells. Concurring with this notion, indomethacin (COX-1/2-inhibitor) was demonstrated to potentiate the cytotoxic effect of IMP extract (GI50 ⦠10.8 µg/ml). The results show that the cytotoxic effect of IMP extract predominates over the influence of proliferative prostanoids released by challenged colorectal cancer cells, and may present a potential source for development of novel anti-cancer drugs.
Asunto(s)
Apoptosis/efectos de los fármacos , Neoplasias Colorrectales/patología , Extractos Vegetales/farmacología , Poaceae/química , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Miembro C3 de la Familia 1 de las Aldo-Ceto Reductasas , Antiinflamatorios no Esteroideos/farmacología , Caspasas/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ciclooxigenasa 1/metabolismo , Inhibidores de la Ciclooxigenasa/farmacología , Dinoprost/metabolismo , Dinoprostona/metabolismo , Células HT29/efectos de los fármacos , Humanos , Hidroxiprostaglandina Deshidrogenasas/metabolismo , Indometacina/farmacología , Componentes Aéreos de las Plantas/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
AIMS: Leydig cells are characterized by their ability to produce testosterone. When the Leydig cells are unable to produce enough testosterone, spermatogenesis fails completely. Considering this, it is of great interest to investigate whether the expressions of steroidogenic enzymes are affected by testicular heat stress. This study aimed to demonstrate that heat induced ER-stress significantly influences steroidogenic enzyme expression and testosterone production in the Leydig cells. MAIN METHODS: C57BL/6 mice were subjected to repetitive testicular heat-treatment at 42 °C for 15 min per day, and heat-treated mLTC-1 cells following hCG treatment for 1h. The protein and RNA expressions were measured by Western blot, RT-PCR. The testosterone and progesterone levels were detected by EIA. The histological and pathological characteristics using hematoxylin and eosin (H&E) and antibody stains. KEY FINDINGS: The 3ß-HSD expression was decreased by heat-stress and hCG treatment. While the GRP78/BiP and CHOP levels were increased by ER-stress inducers, those of the steroidogenic enzyme and progesterone were decreased. In contrast, an ER-stress inhibitor rescued the testosterone levels, even under heat-stress conditions. Moreover, the Leydig cells were randomly scattered, and severely damaged upon repetitive testicular heat-treatment. Additionally, immunohistochemical analyses revealed that cleaved caspase-3 was elevated in the testicular Leydig cells, and rescued by TUDCA. Thus, repetitive testicular heat-treatment in mice promotes excessive ER-stress, thereby leading to apoptosis of the Leydig cells and thus, decreased testosterone production. SIGNIFICANCE: Our findings help to provide an ER-stress mediate mechanistic explanation to the impairment of spermatogenesis upon elevation of the testicular temperature.
Asunto(s)
Estrés del Retículo Endoplásmico , Hipertermia Inducida , Tumor de Células de Leydig/metabolismo , Testosterona/biosíntesis , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Línea Celular Tumoral , Gonadotropina Coriónica/farmacología , Chaperón BiP del Retículo Endoplásmico , Proteínas de Choque Térmico/biosíntesis , Calor , Masculino , Ratones , Ratones Endogámicos C57BL , Progesterona/biosíntesis , ARN/biosíntesis , Esteroides/biosíntesis , Factor de Transcripción CHOP/biosíntesisRESUMEN
This study investigated the effect of resistant maltodextrin (RMD) on reproduction in streptozotocin (STZ)-nicotinamide-induced type 2 diabetic male rats. Forty male rats were induced with diabetes by a single intraperitoneal injection of STZ (50 mg kg(-1)) and nicotinamide (100 mg kg(-1)). Five groups were analysed in total: normal, diabetic rats without RMD, diabetic rats with RMD 1.2 g per 100 g diet (1×), with RMD 2.4 g per 100 g (2×), and with RMD 6.0 g per 100 g (5×). The groups of diabetic rats with the RMD supplement, compared to those without supplement, showed improved plasma glucose control, attenuated insulin resistance and recovery of testosterone level and spermatogenesis stage. The STZ-nicotinamide-induced diabetes mellitus (DM) caused a significant reduction in serum testosterone, testis androgen receptor (AR), steroidogenic acute regulatory protein (StAR) and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) protein, but a statistical recovery in each of these was observed in the 5× group. TUNEL-positive cells were observed in the diabetic without RMD group, and RMD treatment reduced apoptotic germ cells. The expression of Bax/Bcl2 was induced in the diabetic group and also significantly reduced in the 5× group. Dietary RMD may improve metabolic control in STZ-nicotinamide-induced diabetic rats and attenuate hyperglycaemia-related impaired male reproduction and testicular function.
Asunto(s)
Diabetes Mellitus Experimental/complicaciones , Hiperglucemia/complicaciones , Polisacáridos/farmacología , Espermatogénesis/efectos de los fármacos , Testículo/efectos de los fármacos , 17-Hidroxiesteroide Deshidrogenasas/sangre , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/inducido químicamente , Células Germinativas/efectos de los fármacos , Hiperglucemia/sangre , Etiquetado Corte-Fin in Situ , Inyecciones Intraperitoneales , Masculino , Niacinamida/administración & dosificación , Niacinamida/toxicidad , Fosfoproteínas/sangre , Polisacáridos/administración & dosificación , Ratas , Ratas Wistar , Receptores Androgénicos/análisis , Estreptozocina/administración & dosificación , Estreptozocina/toxicidad , Testículo/metabolismo , Testosterona/sangreRESUMEN
Scrotal hyperthermia leads to oxidative stress and apoptosis in spermatogenic cells, which subsequently causes male infertility. In this study, we examined the effects of ß-carotene and/or curcumin on heat-stress- (HS-) induced testicular injuries in mice. ICR male mice (8 weeks old) were consecutively treated with ß-carotene (10 mg/kg) and/or curcumin (20 mg/kg) orally once a day for 14 days and then subjected to single exposure with scrotal HS at 43°C for 15 min on day 7. HS induced a significant reduction in testicular weight, appearance of multinucleated giant cells, and desquamation of germ cells in destructive seminiferous tubules, as well as degenerative Leydig cells. Moreover, HS reduced the superoxide dismutase (SOD) activity and mRNA levels of mitochondrial SOD, phospholipid hydroperoxide glutathione peroxidase, B-cell lymphoma-extra-large, and 3ß-hydroxysteroid dehydrogenase, with increases in lipid peroxidation levels and mRNA levels of BCL2-associated X protein and caspase-3 relative to those of the control group. However, these changes were significantly recovered by combined treatment with ß-carotene and curcumin after HS. These findings indicate that the combined treatment with ß-carotene and curcumin might be a valuable protective agent to ameliorate hyperthermic spermatogenic disorders via its potent antioxidative, antiapoptotic, and androgen synthetic effects.