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1.
本试验旨在研究维生素E对绵羊原代睾丸间质细胞睾酮合成的影响。选择2月龄杜×寒杂交公羔,屠宰后取睾丸组织。分离绵羊睾丸间质细胞,随机分为4个处理组,每个处理组3皿,培养基内添加不同浓度维生素E(0、40、80、160μg/m L)。培养结束后,测定培养基上清睾酮含量,将细胞裂解测定睾酮合成相关酶3β-羟基类固醇脱氢酶(3β-HSD)、17β-羟基类固醇脱氢酶(17β-HSD)、胆固醇侧链裂解酶(P450scc)、17α-羟化酶(CYP17)以及睾酮合成相关基因3β-HSD、17β-HSD、P450scc、CYP17、类固醇合成急性调节蛋白(St AR)基因相对表达量。结果表明:与对照组相比,添加40μg/m L维生素E有提高绵羊睾丸间质细胞睾酮分泌量的趋势(P=0.061),添加40μg/m L维生素E能显著提高P450scc和3β-HSD酶含量(P0.05),P450scc m RNA与3β-HSD m RNA相对表达量也显著提高(P0.05)。 相似文献
2.
试验旨在研究精索上神经（superior spermatic nerve,SSN）对睾丸功能的影响,25只成年Wistar大鼠随机分为2组,试验组左右睾丸切除精索上神经,手术30 d后两组大鼠同时处死取样分析。结果显示,睾丸去精索上神经不影响睾丸指数的大小但使附睾尾精子数极显著降低（P＜0.01）,平均日增重显著增加（P＜0.05）。增殖细胞核抗原（PCNA）免疫组织化学染色结果显示,睾丸去精索上神经不影响曲细精管精原细胞和初级精母细胞的增殖,但影响曲细精管的形态和生殖细胞的规则排列。RT-PCR结果显示,睾丸去精索上神经明显上调β1AR mRNA的表达,下调β2AR mRNA的表达;另外切除精索上神经不影响睾丸3β-羟胆固醇脱氢酶（3β-HSD）mRNA表达,但显著抑制类固醇快速调节蛋白（StAR）、胆固醇侧链裂解酶（P450scc）mRNA表达（P＜0.05）。结果表明,支配睾丸的精索上神经通过调节肾上腺素能受体β1AR和β2AR的表达及StAR和P450scc影响睾丸睾酮的合成和精子的发生。 相似文献
3.
利用RT-PCR和Western-blotting技术分析了胆固醇侧链裂解酶（P450scc）、3β-羟甾脱氢酶（3β-HSD）、C17-20裂解酶（P450c17）、StAR mRNA和蛋白在成年昆明小白鼠心脏、脾、肝、肾中的表达.结果显示,StAR mRNA及其蛋白、P450scc mRNA在以上组织中均有表达;3β-HSD在肾中有较强的表达,在其他组织的表达较弱;P450c17在上述组织中无明显的表达,但在睾丸中有明显的表达.这表明,心脏、脾、肝、肾均具有合成类固醇的能力,但合成类固醇的类型存在一定的差异. 相似文献
4.
牛磺酸对大鼠睾丸间质细胞分泌睾酮的影响及作用机理初探 总被引：2，自引：0，他引：2
将雄性Wistar大鼠（220～250g）的睾丸间质细胞混悬液用不连续梯度（5％、30％、58％、70％）Percoll分离液分离，在细胞培养液中分别加入牛磺酸及促性腺激素（HCG）、雄烯二酮、孕酮、cAMP进行培养，用放射免疫分析方法测定作用24h后培养液中睾酮含量。结果表明，牛磺酸可使睾丸间质细胞分泌睾酮的能力显著增加；能增加HCG和cAMP诱导的睾酮分泌；能促进雄烯二酮和孕酮向睾酮的转化。牛磺酸能增加睾丸间质细胞睾酮的分泌，并可能在类固醇合成的多个环节中都起到了一定的促进作用。 相似文献
5.
高效氯氰菊酯通过ERK1/2影响小鼠睾丸睾酮合成 总被引：2，自引：0，他引：2
为了阐明氯氰菊酯（CP）影响睾酮合成的机制,本研究以成年SPF昆白小鼠为试验动物,通过连续7 d灌喂不同浓度的高效氯氰菊酯（β-CP）,采用RT-PCR、Western blot和放射免疫分析技术分析了β-CP对睾丸重量、血浆类固醇水平、类固醇合成酶、StAR蛋白以及ERK1/2活性的影响。结果显示：随着β-CP浓度的增加,小鼠睾丸的重量无明显变化,但小鼠血浆睾酮的水平逐渐降低;P450sccmRNA、3β-HSDmRNA和StAR mRNA的水平则无明显变化,而P450 c17 mRNA的表达明显下降;StAR蛋白的水平也呈下降趋势,但ERK1/2活性则逐渐升高。以上结果表明,β-CP通过激活ERK1/2而级联抑制P450 c 17 mRNA的转录和StAR前体蛋白的裂解,从而降低睾酮的合成。 相似文献
6.
类固醇激素合成过程中,需要急性调节蛋白(StAR)将细胞质中的胆固醇转运到线粒体中,而StAR基因的表达及相关反应物通过调节细胞中的StAR水平,来调控类固醇激素的合成过程.本文结合大量相关文献资料,首先综述了当前研究者对急性调节蛋白(StAR)的分子结构、编码基因及同源性的研究成果,包括StAR的分子质量、分类类型和同源蛋白转运作用等.随后综述了StAR表达的各种影响因素,包括cAMP-蛋白激酶A(PKA)信号通路、蛋白激酶C(PKC)、调节因子StAR基因及其他因素.一些影响因素是StAR基因转录或转录后表达修饰的参与成分,在影响类固醇激素合成上存在直接、间接作用,在影响作用上存在正向、负向的调节,在影响目标上存在维持稳定、快速增强和抑制降低等类型.最后,探讨了StAR调节类固醇激素合成的作用及其机制,包括跨膜转运胆固醇、StAR对类固醇激素合成的促进作用、巨噬细胞影响StAR调节类固醇生物合成作用.这些作用都可能与机体细胞合成和分泌类固醇激素的水平紧密相关,对动物和人类的生殖内分泌产生很大影响. 相似文献
7.
IGF-Ⅰ对山羊睾丸间质细胞分泌睾酮的调节机理 总被引：2，自引：0，他引：2
以３０～４０日龄山羊的睾丸间质细胞为研究对象,采用体外细胞培养的方法,用胰岛素样生长因子－Ⅰ（ＩＧＦ－Ⅰ）和ｈＣＧ进行不同的处理,用放射免疫分析法（ＲＩＡ）测定了睾酮和ｃＡＭＰ的含量,用荧光分光光度分析法测定了ＤＮＡ含量。结果表明,ＩＧＦ－Ⅰ能促进体外培养的山羊睾丸间质细胞ｃＡＭＰ生成、睾酮分泌及ＤＮＡ合成,以上作用随ＩＧＦ－Ⅰ剂量的增大而加强,并且与ｈＣＧ有协同作用。根据以上结果首次提出ＩＧＦ－Ⅰ调节山羊睾丸间质细胞分泌睾酮机理的假设：ＩＧＦ－Ⅰ与其膜受体结合引起ｃＡＭＰ增多,ｃＡＭＰ通过激活一系列合成ＤＮＡ和睾酮所需酶类而使ＤＮＡ和睾酮的合成增加。这与催乳素（ＰＲＬ）和白细胞介素－１α（ＩＬ－１α）等含氮激素（因子）调节间质细胞分泌睾酮的机理不同,而与ＦＳＨ、ＬＨ等含氮激素调节性腺分泌类固醇激素的作用机理相似。 相似文献
8.
《中国兽医学报》2015,(10):1684-1688
睾酮是一种重要的类固醇激素,动物体内约95%的睾酮都由睾丸间质细胞合成并分泌。研究显示,原癌基因参与性腺功能调控,c-Fos作为原癌基因的1种,在生精细胞发育过程和睾丸内分泌功能中起着重要调控作用。本试验以荣昌仔猪为试验对象,探究c-Fos调节hCG诱导仔猪睾丸间质细胞睾酮分泌的信号转导途径,为进一步研究仔猪睾酮分泌的机理奠定基础。结果显示:(1)间质细胞cAMP在hCG诱导后显著增高,当二丁酰cAMP与hCG共同作用时,睾酮分泌达到最大,且当二丁酰cAMP与c-Fos ASONDs作用时,可明显逆转c-Fos ASONDs对hCG诱导仔猪间质细胞睾酮分泌的抑制作用。(2)维拉帕米(10-5 mol/L)对hCG诱导的间质细胞睾酮分泌未见明显作用,当维拉帕米和c-Fos ASONDs共同作用时也没有加强c-Fos ASONDs对hCG诱导的睾酮分泌的抑制作用。 相似文献
9.
为建立成年小鼠睾丸间质细胞分离纯化方法,并对分离纯化的睾丸间质细胞进行睾酮分泌功能检测,对成年小鼠睾丸组织进行Ⅰ型胶原酶消化、Percoll分离液密度梯度离心,分离成年小鼠睾丸间质细胞.细胞纯度用3β-羟类固醇脱氢酶(3β-hydroxy-steroid-dehydrogenase,3β-HSD)染色鉴定.将分离纯化的睾丸间质细胞进行体外培养,在基础睾酮和人绒毛膜促性腺激素(hCG)刺激培养条件下,用放射免疫分析法对培养上清中的睾酮含量进行检测.结果显示,通过该方法能获得高纯度成年小鼠睾丸间质细胞(＞95％),培养的睾丸间质细胞具有分泌基础睾酮以及对hCG刺激反应的能力.提示采用该方法对成年小鼠睾丸间质细胞进行分离纯化具有高效性、可用性,通过该方法获得的睾丸间质细胞是体外研究药物对睾酮分泌功能影响的良好模型. 相似文献
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Sexual differentiation is a carefully regulated process that ultimately results in a development of the male or female phenotype. Proper development of the male phenotype is dependent upon the action of testosterone and anti‐mullerian hormone. Leydig cells start to produce testosterone around day 12.5 in the fetal mouse testis, and continue to produce high levels of this hormone throughout gestation. In the present study, we examined whether expression of lanosterol 14α‐demethylase (cyp51) and cytochrome P450 NADPH reductase, both involved in the cholesterol production, occurs simultaneously with proteins required for the production of steroid hormones. Immunocytochemical staining with the antibodies against cyp51, cytochrome P450 NADPH reductase, steroidogenic acute regulatory protein (StAR) and 3beta‐hydroxysteroid dehydrogenase I (3β‐HSD I) was used to determine the ontogeny of expression of these four proteins. As expected, 3β‐HSD I and StAR proteins were detected on day 12.5 p.c., while expression of cyp51 and NADPH cytochrome P450 reductase appeared 1 day later, on day 13.5. Thereafter, the expression of all four proteins remained strong throughout gestation. Results of this study suggest that initial steps of steroid hormone production in murine Leydig cells are mostly dependent on exogenously derived cholesterol, while from day 13.5 onwards, mouse Leydig cells are able to synthesize cholesterol and are therefore not dependent on exogenous cholesterol resources. 相似文献
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Weng Q Medan MS Watanabe G Tsubota T Tanioka Y Taya K 《The Journal of reproduction and development》2005,51(3):299-304
The objective of this study was to investigate immunolocalization of steroidogenic enzymes in G?ttingen miniature (GM) pig testes. Testes of 6 adult GM pigs were obtained in September 1996 (n=2), February (n=2) and June (n=2), 1997. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17), and human placental aromatase cytochrome P450 (P450arom). Histologically, all types of spermatogenic cells including mature-phase spermatozoa in seminiferous tubules were observed in all testes throughout the year. Moreover, P450scc, 3betaHSD, P450c17and P450arom were identified in Leydig cells but not in Sertoli cells of all testes. These results suggested that adult GM pig testes have the ability to produce germ cells throughout the year, and the synthesis of progestin, androgen and estrogen occurs in the Leydig cells of GM pig testes. 相似文献
14.
Okano T Murase T Tsubota T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(10):1093-1099
Twenty-one wild male Japanese black bears (Ursus thibetanus japonicus) were captured in the summer-autumn of 1998-2000 in the vicinity of Neo Village, Gifu Prefecture. Testes were measured, and testicular samples were biopsied and observed histologically. Four steroidogenic enzymes, i.e., cholesterol side-chain cleavage cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3betaHSD), 17-alpha hydroxylase cytochrome P450 (P450c17), and aromatase cytochrome P450 (P450arom) were immunolocalized. Serum testosterone concentrations were measured by radioimmunoassay. Testis size changed little from 1-3 years of age, increased rapidly at 4 years, and attained its peak at 5 years. Serum testosterone concentrations ranged from 0.05 to 1.78 ng/m l, and the mean +/- standard deviation was 0.43 +/- 0.48 ng/ml. Age of sexual maturation in wild male Japanese black bears was estimated to be 3-4 years. Seasonal changes in spermatogenesis were obvious; active in June, July and August, degenerated by September. Leydig cells, Sertoli cells and germ cells have the capability of synthesizing androgen, and Leydig cells, Sertoli cells, spermatids and spermatogonia have the capability of synthesizing estrogen in Japanese black bears. 相似文献
15.
Qiang W Murase T Tsubota T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(10):1087-1092
Testes of 15 wild adult male raccoon dogs (Nyctereutes procynoides) obtained from September 2000 to April 2001 were studied to clarify seasonal changes in spermatogenesis and testicular steroidogenesis. There were marked seasonal variations in the testis weight and size with values relatively low in September and highest in March. Spermatogonia and primary spermatocytes were observed in September, while spermatogonia, spermatocytes and round spermatids were present in January, and all types of spermatogenic cells including mature spermatozoa were found in the mating season (February and March). The number of spermatogenic cells reached their peak values in February and March. In addition, steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3 betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17), and human placental aromatase cytochrome P450 (P450arom). P450scc and P450c17 were identified in Leydig cells and spermatids in February, whereas these enzymes were present only in Leydig cells in September. 3betaHSD was found in Leydig cells in September and February with more intense staining in February. The localization of P450arom changed seasonally: no immunostaining in September; more extensive immunostaining in Leydig cells, Sertoli cells, and elongating spermatids in February. These results suggest that seasonal changes in the testis weight and size of wild male raccoon dogs are correlated with changes in spermatogenesis. Seasonal changes in testicular steroidogenesis suggest that the synthesis of androgen and estrogen reaches its peak in the mating season. 相似文献
16.
Previous studies demonstrated significantly lower plasma cortisol level in homozygous halothane-positive (Hal nn) pigs, as compared with homozygous halothane-negative (Hal NN) pigs. To determine whether such difference is attributed to the fundamental alterations in adrenocortical function, F1 offsprings from Pietrain (Hal nn)xErhualian (Hal NN) were intercrossed to produce F2 sibling pigs with segregated genotypes. Adrenocortical cells were isolated from the Hal nn and Hal NN F2 pigs, respectively, and cultured with or without ACTH challenge. Cortisol levels in culture medium, as well as the content of MC2R, cAMP, CREB, phosphorylated CREB (pCREB), StAR and P450scc in adrenocortical cell lysates, were determined. Cortisol, cAMP, StAR and P450scc levels were significantly lower in Hal nn adrenocortical cells under basal condition without ACTH challenge. ACTH significantly increased cortisol level in the medium and the protein content of MC2R, StAR, P450scc in adrenocortical cell lysates, regardless of genotypes. Total CREB protein content was not different between genotypes and treatments, whereas pCREB content exhibited significant effects of genotype and treatment, being higher in Hal NN than in Hal nn under basal condition and in response to ACTH challenge. These results indicate that the compromised cAMP/PKA/pCREB signaling pathway of ACTH and diminished expression of limiting factors in adrenocortical steroidogenesis (StAR and P450scc) may contribute to the significantly lower plasma cortisol levels in Hal nn pigs. 相似文献
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Warita K Sugawara T Yue ZP Tsukahara S Mutoh K Hasegawa Y Kitagawa H Mori C Hoshi N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(12):1257-1267
For the purpose of investigation of working mechanisms in endocrine disruptors, we evaluated the dose-related effects of fetal and/or neonatal exposure to an estrogenic compound on the male reproductive organs in adult mice, particularly with respect to gene expression of steroidogenic acute regulatory protein (StAR). The pregnant ICR mice were given subcutaneous injections of 10 micro g/day/animal of diethylstilbestrol (DES) to subject the fetal mice to in utero exposure (IUE). Subsequently, the newborn male mice were subjected to neonatal exposure (NE) by treatment with vehicle or 0.1-10 micro g/day/animal of DES. Fertility rates of each group were as follows: control, 100%; IUE only, 60%; IUE+NE 0.1 micro g, 25%; IUE+NE 1 micro g, 0%; IUE+NE 10 micro g, 0%. In general histology, germ cell layers in the seminiferous tubules were thinned in the group of IUE+NE 10 micro g. Hypoplasia of the Leydig cells, in which the staining intensity of eosin was diminished, was also observed in the groups of IUE+NE 0.1-10 micro g. The androgen receptor (AR) and estrogen receptor alpha (ERalpha) immunoexpression in the Leydig cells of IUE+NE 1-10 micro g was slightly lower than that in the controls. Long-term dysfunction of the hypothalamo-pituitary-testicular axis, including sustained hypoproduction of gonadotropin and testosterone, and altered expressions of steroid hormone receptors and StAR genes were observed. The hypothalamo-pituitary control of gonadotropin secretion may be affected by the smaller doses of estrogenic agents than the reproductive organs. Furthermore, the fertility rate in the male mice exposed to this estrogenic agent was closely correlated with the testosterone levels, and even more so with the rate-limiting factor of steroidogenesis, StAR. This finding suggests that endocrine disruptors have an important pronounced effect on StAR gene expression. 相似文献
18.
Hayakawa D Sasaki M Akabane C Kitamura N Tsubota T Suzuki M Yamada J 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(11):1463-1466
The testes from 15 adult male Hokkaido Sika deer (Cervus nippon yesoensis) were collected during the rutting season (October and November). We investigated the localization of 4 kinds of steroidogenic enzymes (P450scc, 3betaHSD, P450c17 and P450arom) immunohistochemically in these testicular samples. The specific immunoreactivities to these enzymes were detected only in the cytoplasm of Leydig cells. This differs to the enzyme distributions reported previously in Japanese black bear, Japanese raccoon dog, Hokkaido brown bear and American black bear, in which the same immunoreactivities were detected in Leydig cells, Sertoli cells and/or spermatogenic cells. The current study suggests that in the testes of the Hokkaido Sika deer, testosterone and estradiol-17beta may be synthesized in the Leydig cells only. 相似文献
19.
Weng Q Medan MS Ren L Watanabe G Tsubota T Taya K 《The Journal of reproduction and development》2005,51(2):247-252
In this study, we performed immunohistochemistry of cholesterol side-chain cleavage cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3betaHSD), cytochrome 17alpha-hydroxylase P450 (P450c17), and cytochrome P450 aromatase (P450arom) in the corpus luteum and placenta of Shiba goats. The aim was to clarify the steroidogenic capability of the corpus luteum and placenta of Shiba goats. Ovaries containing corpora lutea were obtained from four adult Shiba goats during the luteal phase (day10; n=2) and pregnancy (90 and 120 days of gestation). Placenta was obtained from one Shiba goat on day 120 of gestation. The sections of the ovaries and placentae were immunostained using the avidin-biotin-peroxidase complex method (ABC) with polyclonal antibodies generated against steroidogenic enzymes of mammalian origin. All luteal cells expressed P450scc, 3betaHSD, P450c17 and P450arom. The distribution of P450scc, 3betaHSD, P450c17 and P450arom were not different during the luteal phase and pregnancy. P450arom showed a weak positive staining in late pregnancy (120 days). In addition, immunoreactions for P450c17 and P450arom were observed in syncytiotrophoblast of the placenta of one Shiba goat. These results indicate that, in Shiba goats, corpus luteum is not only an important source of progesterone but also has the ability to synthesize androgen and estrogen during the luteal phase and pregnancy. Also the placenta has the ability to synthesize androgen and estrogen in late pregnancy. 相似文献