Comparison of cell types in the rat Leydig cell lineage after ethane dimethanesulfonate treatment
Comparison of cell types in the rat Leydig cell lineage after ethane dimethanesulfonate treatment
复制标题
乙烷二甲磺酸盐处理后大鼠 Leydig 细胞谱系的细胞类型比较。
DOI：
10.1530/rep-12-0465
复制
发表时间：
2013-04-01
期刊：
影响因子：
3.8
通讯作者：
Ge, Ren-Shan
中科院分区：
文献类型：
--
作者：
Guo, Jingjing;Zhou, Hongyu;Ge, Ren-Shan
本研究的目的是在乙烷二甲基磺酸盐（EDS）处理后的再生过程中纯化睾丸间质细胞系中的细胞，并比较它们产生类固醇的能力。分别在EDS处理后21天、28天和56天从睾丸中分离出再生祖细胞（RPLCs）、未成熟细胞（RILCs）和成年睾丸间质细胞（RALCs）。在含有100 ng/ml促黄体生成素（LH）的培养基中体外培养3小时后，测量RPLCs、RILCs和RALCs中雄激素（包括雄酮、5α - 雄烷 - 17β，3α - 二醇（DIOL）、睾酮和雄烯二酮）的生成率。在新鲜分离的细胞中测量类固醇生成酶的稳态mRNA水平及其活性。与从正常90日龄大鼠睾丸中分离出的成年睾丸间质细胞（ALCs）（主要产生睾酮（69.73%））相比，RPLCs和RILCs分别主要产生雄酮（70.21%）和DIOL（69.79%）。从EDS处理后56天的睾丸中分离出的睾丸间质细胞显示出与ALCs相当的类固醇生成能力，且主要产生睾酮（72.90%）。RPLCs具有胆固醇侧链裂解酶、3β - 羟基类固醇脱氢酶1和17α - 羟化酶，但几乎检测不到17β - 羟基类固醇脱氢酶3和11β - 羟基类固醇脱氢酶1的活性，而RILCs的17β - 羟基类固醇脱氢酶3和11β - 羟基类固醇脱氢酶1活性增加。由于RPLCs和RILCs具有较高的5α - 还原酶1和3α - 羟基类固醇脱氢酶活性，它们主要产生5α - 还原雄激素。实时定量PCR证实了这些类固醇生成酶表达的类似趋势。总之，纯化的RPLCs、RILCs和RALCs与大鼠青春期发育过程中的相应细胞相似。
The objective of this study was to purify cells in the Leydig cell lineage following regeneration after ethane dimethanesulfonate (EDS) treatment and compare their steroidogenic capacity. Regenerated progenitor (RPLCs), immature (RILCs), and adult Leydig cells (RALCs) were isolated from testes 21, 28 and 56 days after EDS treatment respectively. Production rates for androgens including androsterone and 5a-androstane-17 beta, 3 alpha-diol (DIOL), testosterone and androstenedione were measured in RPLCs, RILCs and RALCs in media after 3-h in vitro culture with 100 ng/ml LH. Steady-state mRNA levels of steroidogenic enzymes and their activities were measured in freshly isolated cells. Compared to adult Leydig cells (ALCs) isolated from normal 90-day-old rat testes, which primarily produce testosterone (69.73%), RPLCs and RILCs primarily produced androsterone (70.21%) and DIOL (69.79%) respectively. Leydig cells isolated from testes 56 days post-EDS showed equivalent capacity of steroidogenesis to ALCs and primarily produced testosterone (72.90%). RPLCs had cholesterol side-chain cleavage enzyme, 3 beta-hydroxysteroid dehydrogenase 1 and 17a-hydroxylase but had almost no detectable 17 beta-hydroxysteroid dehydrogenase 3 and 11 beta-hydroxysteroid dehydrogenase 1 activities, while RILCs had increased 17 beta-hydroxysteroid dehydrogenase 3 and 11 beta-hydroxysteroid dehydrogenase 1 activities. Because RPLCs and RILCs had higher 5 alpha-reductase 1 and 3a-hydroxysteroid dehydrogenase activities they produced mainly 5 alpha-reduced androgens. Real-time PCR confirmed the similar trends for the expressions of these steroidogenic enzymes. In conclusion, the purified RPLCs, RILCs and RALCs are similar to those of their counterparts during rat pubertal development.