The anti-androgenic effects of cypermethrin mediated by non-classical testosterone pathway activation of mitogen-activated protein kinase cascade in mouse Sertoli cells
The anti-androgenic effects of cypermethrin mediated by non-classical testosterone pathway activation of mitogen-activated protein kinase cascade in mouse Sertoli cells
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小鼠支持细胞中丝裂原激活蛋白激酶级联的非经典睾酮途径激活介导氯氰菊酯的抗雄激素作用
DOI：
10.1016/j.ecoenv.2019.03.109
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发表时间：
2019
影响因子：
6.8
通讯作者：
Xu Li Chun
中科院分区：
文献类型：
--
作者：
Wang Qi;Wang Heng Xue;Shen Jun Yu;Zhang Rui;Hong Jia Wei;Li Zheng;Chen Gang;Li Meng Xue;Ding Zhen;Li Jing;Zhang Jin Peng;Zhang Mei Rong;Xu Li Chun
研究表明，氯氰菊酯的抗雄激素作用与睾酮（T）相关的信号通路有关。本研究旨在探讨雷公藤多甙对小鼠睾丸支持细胞（TM 4）的作用，并探讨其作用机制是否通过非经典T细胞信号通路激活丝裂原活化蛋白激酶（MAPK）级联反应而实现。采用细胞计数试剂盒8（CCK 8）和实时细胞分析iCELLigence（RTCA-iCELLigence）系统检测10 μM、20 μM、40 μM和80 μM的顺铂对TM 4细胞活力和增殖的影响。采用哺乳动物双杂交技术、实时荧光定量PCR（qRT-PCR）和western blot技术对T细胞MAPK信号通路中的关键基因和蛋白进行分析。结果表明，MTT能抑制TM 4细胞的增殖和存活。此外，睾酮通过抑制睾酮结合蛋白B（INH B）的表达，促进雄激素结合蛋白（ABP）和转铁蛋白（TF）的表达，干扰支持细胞的功能。此外，RP 1抑制AR和Src激酶的相互作用，并抑制雄激素介导的Src、表皮生长因子受体（EGFR）、细胞外调节激酶1/2（ERK 1/2）和转录因子cAMP反应元件结合蛋白（CREB）的磷酸化。此外，经CYP处理后，雄激素诱导的CREB调节基因早期生长反应因子（Egr 1）的mRNA和蛋白表达下降。结果表明，雷公藤多甙可抑制睾丸支持细胞的活力和增殖，其抗雄激素作用可能与激活MAPK级联的非经典T信号通路有关。这项研究为CYP诱导的生殖毒性提供了新的见解。
Previous studies have demonstrated that the anti-androgenic effects of cypermethrin (CYP) are associated with testosterone (T) - related signaling pathway. This study was to investigate the effects of CYP on mouse Sertoli cells (TM4) and clarify whether the mechanisms were mediated by non-classical T signaling pathway activating mitogen-activated protein kinase (MAPK) cascade. The Cell Counting Kit 8 (CCK8) and Real-Time Cell Analysis iCELLigence (RTCA-iCELLigence) system were performed to detect the effects of 10 μM, 20 μM, 40 μM and 80 μM CYP on the viability and proliferation of TM4. The mammalian two hybrid assay, quantitative Real-Time PCR (qRT-PCR) and western blot were conducted to analyze the key genes and proteins involved in T-mediated MAPK signaling pathway. CYP was found to inhibit the viability and proliferation of TM4. Additionally, CYP disturbed the functions of Sertoli cells by inhibiting inhibin B (INH B) expression and facilitating androgen binding protein (ABP) and transferrin (TF) expression. Moreover, CYP suppressed the interaction of AR and Src kinase and inhibited androgen-mediated phosphorylation of Src, epidermal growth factor receptor (EGFR), extracellular-regulated kinase1/2 (ERK1/2) and transcription factor cAMP response element binding protein (CREB). Furthermore, the androgen-induced mRNA and protein expression of CREB-regulated gene early growth response factor (Egr1) decreased after treated with CYP. It is indicated that CYP inhibits the viability and proliferation of Sertoli cells and non-classical T signaling pathway activation of MAPK cascade is involved in anti-androgenic effect of CYP. This study provides a novel insight into the CYP-induced reproductive toxicity.