铅对睾丸间质瘤细胞孕酮合成能力的影响

发布时间：2018-03-04 00:37

本文选题：铅　切入点：睾丸间质瘤细胞　出处：《毒理学杂志》2017年02期 　论文类型：期刊论文

【摘要】：目的通过检测孕酮合成相关酶的基因表达水平探讨重金属铅对睾丸间质瘤细胞的毒理作用,为揭示重金属导致男性性功能低下的毒性作用机制提供实验依据。方法采用睾丸间质细胞的肿瘤细胞株R2C作为受试对象,暴露于含重金属铅的无血清培养环境,在1、3、6、12和24 h测定细胞活性,收集细胞上清液,利用放射免疫法,检测孕酮合成的变化;应用RT-RCR的方法观察孕酮合成相关酶在基因水平上的表达差异。结果通过CCK-8检测发现,100μmol/L重金属铅作用于R2C细胞12和24 h后,细胞存活率未显著降低(P0.05);放射免疫检测各个时间点孕酮的变化发现,12和24 h能够显著降低孕酮的合成(P0.05)。细胞暴露于含铅培养基后,尽管在作用12和24 h后类固醇激素合成酶基因水平的表达都基本恢复到正常水平,但是在铅暴露3 h后,St AR、P450scc基因的表达显著降低(P0.05),6 h后St AR基因的表达降低更明显(P0.01)。结论重金属铅能够抑制R2C细胞合成孕酮的功能,主要通过下调孕酮合成通路中胆固醇转运酶St AR和限速酶P450scc的基因表达水平发挥作用。
[Abstract]:Objective to investigate the toxic effect of heavy metal lead on testicular stromal tumor cells by detecting the gene expression level of progesterone synthase. Methods the tumor cell line R2C of testicular interstitial cells was used as the object of study, and exposed to the serum-free culture environment containing heavy metal lead. The cell activity was measured at 12 and 24 hours after the treatment. The supernatants were collected and the changes of progesterone synthesis were detected by radioimmunoassay. RT-RCR method was used to observe the difference of progesterone synthase expression at the gene level. Results after 12 and 24 hours of exposure to lead in R2C cells, 100 渭 mol/L heavy metal lead was detected by CCK-8 assay. The cell survival rate was not significantly decreased by radioimmunoassay, and the changes of progesterone at different time points showed that the synthesis of progesterone was significantly decreased at 12 and 24 h. The cells were exposed to lead medium. The expression of steroid hormone synthetase gene returned to normal level after 12 and 24 h exposure. But after 3 h of lead exposure, the expression of St ARN P450scc gene was significantly decreased in R2C cells. Conclusion the heavy metal lead can inhibit the production of progesterone in R2C cells. The expression level of cholesterol transporter St AR and rate-limiting enzyme P450 SCC were down-regulated in progesterone synthesis pathway.
【作者单位】：浙江省金华市中心医院药剂科;浙江省金华市中心医院耳鼻咽喉头颈外科;广州暨南大学生命科学技术学院细胞生物学系生物医药研究院;
【分类号】：R114

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