本文選題：鉛　切入點：睪丸間質(zhì)瘤細(xì)胞　出處：《毒理學(xué)雜志》2017年02期 　論文類型：期刊論文

【摘要】：目的通過檢測孕酮合成相關(guān)酶的基因表達(dá)水平探討重金屬鉛對睪丸間質(zhì)瘤細(xì)胞的毒理作用,為揭示重金屬導(dǎo)致男性性功能低下的毒性作用機制提供實驗依據(jù)。方法采用睪丸間質(zhì)細(xì)胞的腫瘤細(xì)胞株R2C作為受試對象,暴露于含重金屬鉛的無血清培養(yǎng)環(huán)境,在1、3、6、12和24 h測定細(xì)胞活性,收集細(xì)胞上清液,利用放射免疫法,檢測孕酮合成的變化;應(yīng)用RT-RCR的方法觀察孕酮合成相關(guān)酶在基因水平上的表達(dá)差異。結(jié)果通過CCK-8檢測發(fā)現(xiàn),100μmol/L重金屬鉛作用于R2C細(xì)胞12和24 h后,細(xì)胞存活率未顯著降低(P0.05);放射免疫檢測各個時間點孕酮的變化發(fā)現(xiàn),12和24 h能夠顯著降低孕酮的合成(P0.05)。細(xì)胞暴露于含鉛培養(yǎng)基后,盡管在作用12和24 h后類固醇激素合成酶基因水平的表達(dá)都基本恢復(fù)到正常水平,但是在鉛暴露3 h后,St AR、P450scc基因的表達(dá)顯著降低(P0.05),6 h后St AR基因的表達(dá)降低更明顯(P0.01)。結(jié)論重金屬鉛能夠抑制R2C細(xì)胞合成孕酮的功能,主要通過下調(diào)孕酮合成通路中膽固醇轉(zhuǎn)運酶St AR和限速酶P450scc的基因表達(dá)水平發(fā)揮作用。
[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.
【作者單位】： 浙江省金華市中心醫(yī)院藥劑科;浙江省金華市中心醫(yī)院耳鼻咽喉頭頸外科;廣州暨南大學(xué)生命科學(xué)技術(shù)學(xué)院細(xì)胞生物學(xué)系生物醫(yī)藥研究院;
【分類號】：R114

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