Ma Xue, Huang Lu-Gang, Dong Qiang
Department of Pediatric Urology, West China Hospital, Sichuan University, Chengdu 610041, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2013 Oct 18;45(5):770-3.
To investigate the effects of ethane dimethane sulfonate (EDS) on fetal Leydig cells (FLCs) in neonatal rats.
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).
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).
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.
研究乙烷二甲磺酸盐(EDS)对新生大鼠胎儿睾丸间质细胞(FLCs)的影响。
本研究将40只雄性新生SD大鼠分为对照组和实验组。实验组3日龄(出生后第3天,PND3)的大鼠腹腔注射单次EDS(75、100和125mg/kg)。在出生后第7天(PND7)采集样本。称量大鼠体重和睾丸重量,并检测血清睾酮水平。一侧睾丸用于组织学分析(3β-羟基类固醇脱氢酶染色),另一侧睾丸冷冻于冰箱用于分子测定(逆转录-聚合酶链反应、蛋白质免疫印迹法)。
与对照组相比,EDS处理后第4天,75、100和125mg/kg实验组血清睾酮显著降低[(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]。同时,在EDS(100mg/kg)处理组新生雄性大鼠睾丸中,EDS处理后成簇的胎儿睾丸间质细胞消失,Hsd3b1和Cyp17a1表达水平降低(P<0.001)。
乙烷二甲磺酸盐可特异性消耗新生大鼠睾丸中的胎儿睾丸间质细胞。因此,我们可以建立胎儿睾丸间质细胞消耗模型,以更多地了解这些睾丸间质细胞。