铅对睾丸间质瘤细胞孕酮合成能力的影响  

The ability of progesterone synthesis influenced by Lead in Leydig tumor cell

作　　者：詹海潮[1] 邹亚平[2] 仝淑花[1] 张齐好[3] 

机构地区：[1]浙江省金华市中心医院药剂科,金华321000 [2]浙江省金华市中心医院耳鼻咽喉头颈外科 [3]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

出　　处：《毒理学杂志》2017年第2期114-117,共4页Journal of Toxicology

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(P>0.05);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(P<0.05)。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(P<0.05),6 h后St AR基因的表达降低更明显(P<0.01)。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。Objective To reveal the toxic mechanism of damage about Androgen deficiency in male and male hypogonadism induced by heavy metal, an experiment through detecting the gene expression level of progesterone synthesis enzymes had offered a proof for the toxic mechanism of damage induced by Lead in Leydig tumor cell. Methods Leydig tumor cell R2C was used as a subject in this study. After exposed to lead with serum-free, cultured for 24h. Cell viability was determined and cell supernatant was collected after 1,3,6,12 and 24h, detected by radioimmunoassay, progesterone changes were uncovered. RT-PCR was applied to make a co-verification on enzymes with the differernees in the level of gene expression during progesterone synthesis pathways. Results Lead exposure caused a significantly decrease in the synthesis of progesterone after 12 and 24 h （P 〈 0.05 ）, but cell growth was no significantly difference between control and test group （P 〉0. 05 ）. Although steroid hormone synthesis enzymes gradually returned to normal levels after 12 and 24 h, the obviously decreasing expression of StAR and P450scc gene after 3 h exposed to Lead with serum-free （ P 〈 0. 05 ）. The expression of StAR gene had been significantly decreased after treatment 6 h（ P 〈 0. 01 ）. Conclusions The synthesis of progesterone had been hampered by Lead and the decreasing expression of StAR and P450sec gene was the mainly reason.

关 键 词：铅 睾丸间质瘤细胞 STAR P450SCC 3Β-HSD 

分 类 号：R994.6[医药卫生—毒理学]