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Objective: To investigate the effects of ethane dimethane sulfonate (EDS) on fetal Leydig cells (FLCs) in neonatal rats., Methods: In the study, 40 male neonatal SD rats were divided into control group and experimental groups. The rats in the experimental groups aged 3 days (PND3) were intraperitoneally injected with one single EDS (75, 100 and 125 mg/kg). The samples were collected on PND7. The body and testes was weighed, and the serum level of testosterone was detected. One testis was for histological analysis (3β-hydroxysteroid dehydrogenase stainining), and the other was frozen in refrigerator for molecular determination (RT-PCR, Western blot)., Results: Compared with the control group, on day 4 after EDS treatment, significant decrease of serum testosterone was observed in the 75, 100 and 125 mg/kg experimental groups [(0.542 ± 0.117) μg/L, (0.124 ± 0.021) μg/L, (0.113 ± 0.015) μg/L, vs. (0.834 ± 0.172) μg/L, P<0.05]. Meanwhile, fetal Leydig cells in clusters disappeared with a lower expression level of Hsd3b1 and Cyp17a1 after EDS treatment in the testes of neonatal male rats in EDS (100 mg/kg)-treatment group (P<0.001)., Conclusion: Ethane dimethane sulfonate can specifically deplete the fetal Leydig cells in testes of neonatal rats. Thus we could establish the FLCs' depletion model to know more about these testicular interstitial cells.

Objective: To explore the feasibility of inducing the differentiation of human adipose tissue-derived stem cells (ADSCs) into Leydig cells in vitro., Methods: We isolated ADSCs by digestion with Collagenase I from the subcutaneous adipose tissue, cultured them in the DMEM/F12 medium with 10% fetal bovine serum, and detected the expression of vimentin by immunohistochemistry. We exposed the ADSCs to different concentrations of human chorionic gonadotropin (HCG) for different times, determined the expression of StAR mRNA by real-time PCR, and measured the HCG-induced proliferation of ADSCs by MTT. After a week of induction by HCG and DMSO, we conducted 3beta-HSD immunohistochemistry, and detected the testosterone level in the supernatant and lysis of the cells by radioimmunoassay., Results: The ADSCs grew well with a positive expression of vimentin. The expression of the StAR gene was positively correlated with the increased concentration of HCG, reaching the peak at HCG 10 U/ml in 1 week culture. The proliferation of ADSCs was significantly increased by HCG induction. A positive expression of 3beta-HSD was observed after 1 week induction with HCG 10 U/ml and DMSO 3.2 x 10(-6)mol/L., Conclusion: HCG enhances the expression of the StAR gene and the proliferation of ADSCs. Induced by HCG 10 U/ml and DMSO 3.2 x 10(-6) mol/L, ADSCs tend to differentiate into Leydig cells.

Objective: To observe the influence of the organophosphate insecticide dichlorvos on the apoptosis of Leydig cells in the male offspring of the SD rats exposed to dichlorvos, and to investigate the role of the changes of Leydig cells in genitourinary malformation., Methods: Twenty-one pregnant SD rats were divided into a corn oil control group and 6 dichlorvos groups, the former given by gavage 1.0 ml corn oil daily, and the latter dichlorvos at the dose of 1, 4, 8, 16, 20 and 24 mg/kg daily from the 12th to 17th day of conception. After birth, 5 male neonates were randomly selected from each of the control and dichlorvos groups, and their testes were harvested to be analyzed by HE staining, immunohistochemistry with anti-caspase-3 antibodies and DAPI fluorescent staining. At 90 days after birth, another 5 of the male offspring were taken from each group and their testes were collected for the same analyses., Results: Statistically significant differences were found in the number of both the caspase-3 positive and DAPI labeled Leydig cells in the testes of the rat offspring between the corn oil and the 4, 8, 16, 20 and 24 mg/kg dichlorvos groups (P < 0.05), but not between the control and the 1 mg/kg dichlorvos groups (P > 0.05). The apoptosis of Leydig cells was increased in the male offspring of the dichlorvos-exposed SD rats in a dose-dependent manner., Conclusion: Exposure of pregnant rats to dichlorvos can increase the apoptosis of Leydig cells in the male offspring, which, in turn, may reduce the number of Leydig cells, interfere with the testis function during the embryonic period, and damage the development of the genitourinary system.