TY - JOUR
T1 - Phthalate inhibits Leydig cell differentiation and promotes adipocyte differentiation
AU - Hao, Xinrui
AU - Guan, Xiaoju
AU - Zhao, Xingxing
AU - Ji, Minpeng
AU - Wen, Xin
AU - Chen, Panpan
AU - Chen, Fenfen
AU - Yang, Jianying
AU - Lian, Qingquan
AU - Ye, Leping
AU - Chen, Haolin
N1 - Funding Information:
This work is supported by grants from Chinese central or local governments, including National Natural Science Foundation of China Grant 91949123 (HC), 81771635 (LY) and Wenzhou City Key Scientific and Technological Innovation Project ZY2019002 (HC). These sources did not influence the study design, the collection, analysis or interpretation of data, or the writing of the paper.
Funding Information:
In addition to steroidogenic genes, MEHP also significantly increased the genes involved in testosterone metabolism (Srd5a and Akr1c14) and testosterone metabolic product DIOL, while inhibited other no-steroidogenic genes (Insl3, Hao2 and Pah). During Leydig cell development, the two testosterone metabolic genes are normally up-regulated first in immature Leydig cells and then down-regulated in adult cells, which corresponds to a transitory increase in the metabolic product DIOL (Ge and Hardy, 1998). The up-regulations of both metabolic genes and the metabolic product DIOL by MEHP at the end of week 3 suggest strongly that MEHP may slow down the differentiation of SLCs. This conclusion is also supported by the down-regulation of three no-steroidogenic genes. Insl3 is an important peptide hormone produced by Leydig cells which plays a critical role in testicular descent in fetal period. The specific role in adult is still unclear but its expression is regarded as an important functional marker for adult Leydig cells (Ivell et al., 2013). Hao2 (2-hydroxyacid oxidase) and Pah (phenylalanine hydroxylase) are another two Leydig cell specific no-steroidogenic genes (O'Shaughnessy et al., 2014). Hao2 encodes a protein localizes to the peroxisome with activity toward the substrate 2-hydroxypalmitate. Pah encodes a protein that is responsible for phenylalanine metabolism. These three genes are involved in biological functions other than steroidogenesis and their expressions are less relied on LH stimulations. Their down-regulations suggest strongly that MEHP may affect the development of the no-steroidogenic functions of Leydig cells. Overall, the down-regulations in both steroidogenic and no-steroidogenic genes and the up-regulations in testosterone metabolic genes, as well as reductions in testosterone production itself support the conclusion that MEHP may inhibit SLC differentiation, though specific acute effects on a few steroidogenic genes may also be present.This work is supported by grants from Chinese central or local governments, including National Natural Science Foundation of China Grant 91949123 (HC), 81771635 (LY) and Wenzhou City Key Scientific and Technological Innovation Project ZY2019002 (HC). These sources did not influence the study design, the collection, analysis or interpretation of data, or the writing of the paper.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2021/1
Y1 - 2021/1
N2 - Studies have shown that phthalates are capable of affecting the development and functions of male reproductive system. The effect of phthalates on Leydig cell functions is well documented. However, little is known about their potential effects on the functions of stem Leydig cells (SLC). In the present study, we have examined the effects of mono-(2-ethylhexyl) phthalate (MEHP) on SLC functions in vitro by culturing seminiferous tubules and isolated SLCs. The results indicate that MEHP can significantly inhibit the proliferation and differentiation of SLCs in both the organ and cell culture systems. Interestingly, the minimal effective concentration that is able to affect SLC function was lower in the tubule culture system (1 μM) than in the isolated cells (10 μM), suggesting a possible involvement of the niche cells. Also, MEHP appeared to affect both the efficiency of SLCs to form Leydig cells and a selected group of Leydig cell-specific genes, including Lhcgr, Scarb1, Hsd3b1, Cyp17a1, Star, Srd5a1, Akr1c14, Insl3, Hao2 and Pah. Since SLCs are multipotent, we also tested the effect of MEHP on the differentiation of SLCs to adipocytes. Though MEHP by itself can not specify SLCs into adipocyte lineage, it indeed significantly increased the adipogenic activity of SLCs if used with an adipocyte inducing medium by up-regulation of multiple adipogenic-related genes, including Pparg and Cebpa. Overall, the results indicate that MEHP inhibits SLCs differentiating into Leydig lineage while stimulates the differentiating potential of SLCs to adipocytes.
AB - Studies have shown that phthalates are capable of affecting the development and functions of male reproductive system. The effect of phthalates on Leydig cell functions is well documented. However, little is known about their potential effects on the functions of stem Leydig cells (SLC). In the present study, we have examined the effects of mono-(2-ethylhexyl) phthalate (MEHP) on SLC functions in vitro by culturing seminiferous tubules and isolated SLCs. The results indicate that MEHP can significantly inhibit the proliferation and differentiation of SLCs in both the organ and cell culture systems. Interestingly, the minimal effective concentration that is able to affect SLC function was lower in the tubule culture system (1 μM) than in the isolated cells (10 μM), suggesting a possible involvement of the niche cells. Also, MEHP appeared to affect both the efficiency of SLCs to form Leydig cells and a selected group of Leydig cell-specific genes, including Lhcgr, Scarb1, Hsd3b1, Cyp17a1, Star, Srd5a1, Akr1c14, Insl3, Hao2 and Pah. Since SLCs are multipotent, we also tested the effect of MEHP on the differentiation of SLCs to adipocytes. Though MEHP by itself can not specify SLCs into adipocyte lineage, it indeed significantly increased the adipogenic activity of SLCs if used with an adipocyte inducing medium by up-regulation of multiple adipogenic-related genes, including Pparg and Cebpa. Overall, the results indicate that MEHP inhibits SLCs differentiating into Leydig lineage while stimulates the differentiating potential of SLCs to adipocytes.
KW - Adipocyte
KW - Phthalate
KW - Stem leydig cell
KW - Steroidogenesis
KW - Testosterone
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U2 - 10.1016/j.chemosphere.2020.127855
DO - 10.1016/j.chemosphere.2020.127855
M3 - Article
C2 - 32799149
AN - SCOPUS:85089274357
VL - 262
JO - Chemosphere
JF - Chemosphere
SN - 0045-6535
M1 - 127855
ER -