1.Expression of 3b-Hydroxysteroid dehydrogenase and P450 side chain cleavage enzyme in the human uterine endometrium.
Hee Sub RHEE ; Seon Hee OH ; Bum Joo KO ; Dong Min HAN ; Byung Hun JEON ; Hyun PARK ; Hyung Bae MOON ; Won Sin KIM
Experimental & Molecular Medicine 2003;35(3):160-166
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.
Cholesterol/chemistry
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Cholesterol Side-Chain Cleavage Enzyme/*biosynthesis/genetics
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Decidua/enzymology
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Endometrium/*enzymology
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Female
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Gene Expression/physiology
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Human
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Menstrual Cycle/physiology
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Multienzyme Complexes/*biosynthesis/genetics
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Placenta/enzymology
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Pregnancy
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Pregnenolone/biosynthesis
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Progesterone/biosynthesis
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Progesterone Reductase/*biosynthesis/genetics
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Steroid Isomerases/*biosynthesis/genetics
2.Influence of electromagnetic irradiation on P450scc mRNA expression in rat testis tissues and protective effect of the shield.
Wen ZHOU ; Xu-bu WANG ; Jin-qing YANG ; Yong LIU ; Guang-bin ZHANG
National Journal of Andrology 2005;11(4):269-271
OBJECTIVETo 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.
METHODSHealthy 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.
RESULTSThe 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.
CONCLUSIONElectromagnetic 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.
Animals ; Cholesterol Side-Chain Cleavage Enzyme ; biosynthesis ; genetics ; Copper ; Male ; RNA, Messenger ; genetics ; Radiation Protection ; instrumentation ; Radioimmunoassay ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Testis ; metabolism ; radiation effects ; Testosterone ; blood
3.Gonadotrophin-releasing hormone-I and -II stimulate steroidogenesis in prepubertal murine Leydig cells in vitro.
Yung-Ming LIN ; Ming-Yie LIU ; Song-Ling POON ; Sew-Fen LEU ; Bu-Miin HUANG
Asian Journal of Andrology 2008;10(6):929-936
AIMTo study the effect and mechanism of gonadotrophin-releasing hormone (GnRH) on murine Leydig cell steroidogenesis.
METHODSPurified murine Leydig cells were treated with GnRH-I and -II agonists, and testosterone production and steroidogenic enzyme expressions were determined.
RESULTSGnRH-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).
CONCLUSIONGnRH directly stimulated murine Leydig cell steroidogenesis by activating 3b-HSD enzyme expression.
3-Hydroxysteroid Dehydrogenases ; biosynthesis ; genetics ; Animals ; Blotting, Western ; Cell Separation ; Cells, Cultured ; Cholesterol Side-Chain Cleavage Enzyme ; biosynthesis ; Dose-Response Relationship, Drug ; Gonadotropin-Releasing Hormone ; agonists ; pharmacology ; Leydig Cells ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phosphoproteins ; biosynthesis ; genetics ; RNA ; biosynthesis ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sexual Maturation ; physiology ; Steroids ; biosynthesis ; Testosterone ; biosynthesis
4.Ginkgo biloba extract enhances testosterone synthesis of Leydig cells in type 2 diabetic rats.
Xiao-Ye WU ; Wen-Yan WANG ; Rong-Rong WANG ; Lin XIE ; Zhou-Xi FANG ; Guo-Rong CHEN
National Journal of Andrology 2008;14(4):371-376
OBJECTIVETo investigate the effects of Ginkgo biloba extract (EGB) on the testosterone synthesis in the Leydig cells of type 2 diabetic rats.
METHODSThirty 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.
RESULTSCompared 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.
CONCLUSIONEGB enhances testosterone synthesis and secretion of Leydig cells by reducing the impairment of the testis in type 2 diabetic rats.
17-Hydroxysteroid Dehydrogenases ; genetics ; Animals ; Cholesterol Side-Chain Cleavage Enzyme ; genetics ; Diabetes Mellitus, Type 2 ; blood ; genetics ; physiopathology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; drug effects ; Ginkgo biloba ; chemistry ; Hydroxysteroid Dehydrogenases ; genetics ; Leydig Cells ; drug effects ; metabolism ; ultrastructure ; Luteinizing Hormone ; blood ; Male ; Microscopy, Electron, Transmission ; Phosphoproteins ; genetics ; Plant Extracts ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; biosynthesis ; blood