The increased number of Leydig cells by di(2-ethylhexyl) phthalate comes from the differentiation of stem cells into Leydig cell lineage in the adult rat testis
The increased number of Leydig cells by di(2-ethylhexyl) phthalate comes from the differentiation of stem cells into Leydig cell lineage in the adult rat testis
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DOI：
10.1016/j.tox.2013.01.021
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发表时间：
2013-04-05
期刊：
影响因子：
4.5
通讯作者：
Ge, Ren-Shan
中科院分区：
文献类型：
--
作者：
Guo, Jingjing;Li, Xing-Wang;Ge, Ren-Shan
本研究的目的是确定在成年期暴露于邻苯二甲酸二（2-乙基己基）酯（DEHP）是否会增加大鼠睾丸间质细胞数量，并探讨可能的机制。将90日龄Long-Evans大鼠随机分为3组，分别灌胃玉米油（对照组）或10或750 mg/kg DEHP，连续7 d，然后腹腔注射75 mg/kg乙烷二甲磺酸盐（EDS）清除Leydig细胞。采用放射免疫法测定血清睾酮浓度，qPCR法测定睾丸间质细胞基因mRNA水平。EDS后第4天，EDS消除了对照睾丸中的所有Leydig细胞，这通过检测不到血清睾酮水平和睾丸中无3 β-羟基类固醇脱氢酶阳性（3 β-HSDpos）细胞来判断。然而，在DEHP处理组中，有可检测的血清睾酮浓度和一些椭圆形的3 β-HSDpos细胞在睾丸。这些3 β-HSDpos细胞未被抗11 β-羟基类固醇脱氢酶1（11 β-HSD 1）的抗体染色，11 β-HSD 1是更晚期Leydig细胞的标志物。EDS后第4天，对照组睾丸中Leydig细胞生物标志物Lhcgr、Cyp 11 a1、Cyp 17 a1、Insl 3和Hsd 11b 1的mRNA消失。然而，在DEHP处理的睾丸中有可检测浓度的Lhcgr、Cyp 11 a1和Cyp 17 a1 mRNA，但未检测到Insl 3、Hsd 17 b3和Hsd 11b 1浓度，表明这3种β-HSDpos细胞是新形成的Leydig祖细胞。EDS后第4天，对照睾丸中巢蛋白（内斯，干间质细胞的生物标志物）的mRNA水平显著增加，但DEHP处理的睾丸中没有，表明这些巢蛋白阳性干细胞在DEHP处理的睾丸中分化为前体间质细胞。目前的研究表明，DEHP可促进干细胞分化为Leydig祖细胞。(C)2013爱思唯尔爱尔兰有限公司版权所有。
The objective of the present study is to determine whether di(2-ethylhexyl) phthalate (DEHP) exposure at adulthood increases rat Leydig cell number and to investigate the possible mechanism. 90-day-old Long-Evans rats were randomly divided into 3 groups, and were gavaged with the corn oil (control) or 10 or 750 mg/kg DEHP daily for 7 days, and then received an intraperitoneal injection of 75 mg/kg ethane dimethanesulfonate (EDS) to eliminate Leydig cells. Serum testosterone concentrations were assessed by RIA, and the mRNA levels of Leydig cell genes were measured by qPCR. EDS eliminated all Leydig cells in the control testis on day 4 post-EDS, as judged by undetectable serum testosterone level and no 3 beta-hydroxysteroid dehydrogenase positive (3 beta-HSDpos) cells in the interstitium. However, in DEHP-treated groups, there were detectable serum testosterone concentrations and some oval-shaped 3 beta-HSDpos cells in the interstitium. These 3 beta-HSDpos cells were not stained by the antibody against 11 beta-hydroxysteroid dehydrogenase 1 (11 beta-HSD1), a marker for Leydig cells at a more advanced stage. The disappearance of mRNAs of Leydig cell biomarkers including Lhcgr, Cyp11a1, Cyp17a1, Insl3 and Hsd11b1 in the control testis was observed on day 4 post-EDS. However, there were detectable concentrations of Lhcgr, Cyp11a1 and Cyp17a1 mRNAs but undetectable concentrations of Insl3, Hsd17b3 and Hsd11b1 in the DEHP-treated testes, indicating that these 3 beta-HSDpos cells were newly formed progenitor Leydig cells. The mRNA level for nestin (Nes, biomarker for stem Leydig cells) was significantly increased in the control testis on day 4 post-EDS, but not in the DEHP treated testes, suggesting that these nestin positive stem cells were differentiated into progenitor Leydig cells in the DEHP-treated testes. The present study suggests that DEHP increases the differentiation of stem cells into progenitor Leydig cells. (C) 2013 Elsevier Ireland Ltd. All rights reserved.