title: 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.
abstract: 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 3β-hydroxysteroid dehydrogenase positive (3β-HSD(pos)) cells in the interstitium. However, in DEHP-treated groups, there were detectable serum testosterone concentrations and some oval-shaped 3β-HSD(pos) cells in the interstitium. These 3β-HSD(pos) cells were not stained by the antibody against 11β-hydroxysteroid dehydrogenase 1 (11β-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 3β-HSD(pos) 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.
authors: Institute of Reproductive Biomedicine and the 2nd Affiliated Hospital, Wenzhou Medical College, Wenzhou, Zhejiang 325000, PR China.
authors: Jingjing
authors: J
authors: Guo
authors: Xing-Wang
authors: XW
authors: Li
authors: Yong
authors: Y
authors: Liang
authors: Yufei
authors: Y
authors: Ge
authors: Xiaomin
authors: X
authors: Chen
authors: Qing-Quan
authors: QQ
authors: Lian
authors: Ren-Shan
authors: RS
authors: Ge
mesh_terms: D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D007985:Leydig Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D008698:Mesylates / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D010968:Plasticizers / Q000633:toxicity*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011897:Random Allocation; D051381:Rats; D020318:Rats, Long-Evans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013739:Testosterone / Q000097:blood
chemical_list: D007381:Intermediate Filament Proteins; D008698:Mesylates; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D010968:Plasticizers; D013739:Testosterone; D012313:RNA; D007986:Luteinizing Hormone; D004051:Diethylhexyl Phthalate; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; C002994:ethylene dimethanesulfonate