Studies on LH modulated 8kDa peptide involved regulation of testosterone production in rat Leydig cells

Aim: To demonstrate the role of the 8 kDa peptide in regulation of testosterone production by mt Leydig cells. Meth-ods: A peptide similar to 8 kDa peptide purified from immature rat Leydig cells was isolated and purified from rat lungcytosol. Immunological and structural similarity between the peptides purified from lung and Leydig cells was estab-lished by Western blot and tryptic map comparison respectively. Results: Addition of the 8 kDa peptide 10, 50, 100;and 150 ug decreased the production of testosterone in Leydig cells dose-dependently. But the addition of the peptide150 ug along with hCG had no effect on hCG-stimulated increase in testosterone production. Conclusion: In vitro ad-dition of the peptide purified from lung cytosol to adult rat Leydig cells resulted in a concentration-dependent decrease inbasal testosterone production although it had no effect on hCG-stimulated testosterone production. (Asian J Androl1999 Dec; 1: 191-194)     

Immunoregulative Mechanism of the Leydig Cell in the Infection by Expression of IL-1,IL-6,TGF-β, Fas and FasL mRNA

Objective To investigate the immune regulative mechanism of Leydig cells in the localinfection of rat's testis.Methods Ureaplasma Urealyticum(UU) was injected into rat's bladder, which mimicked an ascending infectious way, and at the same time culture medium was injected into rat's bladder as the control. The rats were sacrificed at week 1, 2 and 3 after injection respectively. Then pathological changes in testis were analyzed by histological examination. At the same time Leydig cells were separated from rat's testis, The comparasion of levels of IL-1,IL-6, TGF-β, Fas and FasL mRNA expression among the three groups was made by RT-PCR.Results As compared with control group, the levels of IL-1, IL-6, TGF-β mRNA expression for UU supernatant and living UU groups increased; and levels of Fas and FasL mRNA expression decreased and increased respectively after UU infection.Conclusion During anti-infective immunity, rat's Leydig cells may regulate immune function of the testis by changing the levels of IL-1, IL-6, TGF-β, Fas and FasL mRNA expression and may contribute to maintain immune privilege of the testis.     

Pure Sertoli cell tumor of the ovary: A case report

Pure Sertoli cell tumors are an uncommon variant of rare ovarian Sertoli‐Leydig cell tumors. Due to nonspecific clinical and imaging features, diagnosis is often made after histopathological examination. The prognosis is excellent as most are detected in the early stages and surgical resection is often curative in most cases.     

THYROID HORMONES: THEIR ROLE IN TESTICULAR STEROIDOGENESIS

Thyroid hormones are important for growth and development of many tissues. Altered thyroid hormone status causes testicular abnormalities. For instance, juvenile hypothyroidism/neonatal transient hypothyroidism induces macroorchidism, increases testicular cell number (Sertoli, Leydig, and germ cells) and daily sperm production. Triiodothyronine (T3) receptors have been identified in sperm, developing germ cells, Sertoli, Leydig, and peritubular cells. T3 stimulates Sertoli cell lactate secretion as well as mRNA expression of inhibin- &#102, androgen receptor, IGF-I, and IGFBP-4. It also inhibits Sertoli cell mRNA expression of Müllerian inhibiting substance (MIS), aromatase, estradiol receptor, and androgen binding protein (ABP) and ABP secretion. T3 directly increases Leydig cell LH receptor numbers and mRNA levels of steroidogenic enzymes and steroidogenic acute regulatory protein. It stimulates basal and LH-induced secretion of progesterone, testosterone, and estradiol by Leydig cells. Steroidogenic factor-1 acts as a mediator for T3-induced Leydig cell steroidogenesis. Although the role of T3 on sperm, germ, and peritubular cells has not yet been completely studied, it is clear that T3 directly regulates Sertoli and Leydig cell functions. Further studies are required to elucidate the direct effect of T3 on sperm, germ, and peritubular cells.     

Nitric oxide production of rat Leydig and Sertoli cells is stimulated by round spermatid factor(s)

In this study, we provide evidence of cell-to-cell interaction between rat germ cells and Leydig or Sertoli cells in relation to nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) expression. As a result of being cultured in a round spermatid-conditioned medium (RSd-CM), NO production in both Leydig and Sertoli cells increased in proportion to the length of the culture period. iNOS mRNA expression in both types of cells also increased in a dose-dependent manner as a result of being cultured with RSd-CM. This increase was detected as early as 3 h and was maintained up to 24 h. In contrast, neither NO production nor iNOS mRNA increased in either type of cell following culture in a pachytene spermatocyte-conditioned medium (PS-CM). Our findings suggest that RSd may control NO production of Leydig and Sertoli cells. This cell-to-cell interaction may be an important mechanism of regulation of testicular function.     

Differential effect of corticosterone deficiency on the expression of LH, prolactin and insulin receptors on rat Leydig cells

The adverse effects of glucocorticoid deficiency on the expression of genes encoding Leydig cell surface receptors and the response to LH/prolactin/insulin to produce testosterone production are yet to be recognized. Following metyrapone-induced corticosterone deficiency, serum corticosterone, testosterone and insulin levels decrease, whereas serum prolactin exhibits a significant increase and serum LH remains unaltered. LH binding and LH receptor mRNA expression were not altered, but a significant decrease in PRL and insulin binding and in the mRNA expressions of their receptors were observed in corticosterone-deficient rats in vivo. Corticosterone deficiency significantly decreases the Leydig cellular basal as well as hormone-stimulated testosterone production in vitro. Simultaneous administration of corticosterone prevented its deficiency-induced changes in Leydig cells both in vivo and in vitro. Our results show that metyrapone-induced corticosterone deficiency impairs Leydig cell insulin and prolactin receptors, and their mRNA expression and the response of Leydig cells to LH/PRL/insulin on testosterone production.     

双酚A对小鼠睾丸间质细胞毒性及miR-203-3p和PI3K/AKT/FOXO1信号通路的影响

目的研究双酚A对小鼠睾丸间质细胞的毒性作用,及对miR-203-3p和PI3K/AKT/FOXO1信号通路的影响。方法不同浓度BPA(0、2、10、50、250μmol/L)处理小鼠睾丸间质细胞24 h,CCK8法检测细胞活力,Real time PCR检测miR-203-3p和FOXO1、AKT、PI3K的相对表达水平。结果不同浓度BPA处理细胞后,细胞活力随BPA剂量的增大而减少,50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。各处理组miR-203-3p表达量均较对照组升高,10、250μmol/L组与对照组比较,差异有统计学意义(P<0.05)。2、10μmol/L组FOXO1表达量较对照组升高,50、250μmol/L组表达量较对照组降低,2、50、250μmol/L组FOXO1相对表达量与对照组比较差异有统计学意义(P<0.001)。各处理组AKT水平均出现下降趋势,10、50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。各处理组PI3K水平均出现下降趋势,50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。结论双酚A致睾丸间质细胞损伤,影响miR-203-3p和FOXO信号通路相关基因的改变。     

Alterations of gene profiles in Leydig-cell-regenerating adult rat testis after ethane dimethane sulfonate-treatment

Only occupying about 1%–5% of total testicular cells, the adult Leydig cell (ALC) is a unique endocrine cell that produces androgens. Rat Leydig cells regenerate after these cells in the testis are eliminated with ethane dimethane sulfonate (EDS). In this study, we have characterized Leydig cell regeneration and messenger ribonucleic acids (mRNA) profiles of EDS treated rat testes. Serum testosterone, testicular gene profiling and some steroidogenesis-related proteins were analyzed at 7, 21, 35 and 90 days after EDS treatment. Testicular testosterone levels declined to undetectable levels until 7 days after treatment and then started to recover. Seven days after treatment, 81 mRNAs were down-regulated greater than or equal to two-fold, with 48 becoming undetectable. These genes increased their expression 21 days and completely returned to normal levels 90 days after treatment. The undetectable genes include steroidogenic pathway proteins: steroidogenic acute regulatory protein, Scarb1, Cyp11a1, Cyp17a1, Hsd3b1, Cyp1b1 and Cyp2a1. Seven days after treatment, there were 89 mRNAs up-regulated two-fold or more including Pkib. These up-regulated mRNAs returned to normal 90 days after treatment. Cyp2a1 did not start to recover until 35 days after treatment, indicating that this gene is only expressed in ALCs not in the precursor cells. Quantitative polymerase chain reaction, western blotting and semi-quantitative immunohistochemical staining using tissue array confirmed the changes of several randomly picked genes and their proteins.     

碱性成纤维细胞生长因子在大鼠BMSCs向睾丸Leydig细胞分化中的作用研究

目的 :研究碱性成纤维细胞生长因子(b FGF)在大鼠骨髓间充质干细胞(BMSCs)定向分化为睾丸Leydig细胞过程中的作用,探讨BMSCs定向分化为Leydig细胞的适宜方法。方法:将经纯化鉴定的SD大鼠BMSCs第3代细胞接种于6孔板,随机分为对照组(A)、人绒毛膜促性腺素(h CG)+血小板源性生长因子(PDGF)诱导组(B)、h CG+PDGF+5.0 ng/ml b FGF诱导组(C)、h CG+PDGF+10.0 ng/ml b FGF诱导组(D)、h CG+PDGF+20.0 ng/ml b FGF诱导组(E),分别于第7、14、21天进行形态学观察、培养液睾酮水平及3β-羟化类固醇脱氢酶(3β-HSD)免疫荧光检测。结果:B、C、D、E组细胞诱导后体积变大,互相连接呈长梭形或不规则形,贴壁生长,核大、较圆,符合Leydig细胞的特点。随着时间和b FGF浓度的增加,B、C、D、E组睾酮水平逐渐升高,3β-HSD阳性表达细胞逐渐增多,C、D、E组与B组,D、E组与C组间差异有统计学意义(P均<0.05);D、E组之间差异无统计学意义(P>0.05)。结论:b FGF在大鼠BMSCs向Leydig细胞体外诱导分化过程中有明显促进作用。