硫酸镍对体外培养的大鼠睾丸间质细胞类固醇合成酶的影响  

Effect of nickel sulfate on steroidogenic enzymes of rat Leydig cells in vitro

作　　者：李坚[1] 胡晓婷[1] 张彩霞[1] 王良[1] 陶熙[1] 苏莉[1] 赵迟[1] 孙应彪[1] 

机构地区：[1]兰州大学公共卫生学院,甘肃兰州730000

出　　处：《工业卫生与职业病》2014年第3期165-168,共4页Industrial Health and Occupational Diseases

基　　金：中央高校基本科研业务费专项资金(Lzujbky-2012-158);国家级大学生创新创业训练计划项目(201210730039)

摘　　要：目的探讨硫酸镍对体外培养的大鼠睾丸间质细胞睾酮(T)分泌及类固醇合成酶基因表达的影响。方法采用体外分离、纯化和培养的大鼠睾丸间质细胞,分别用0、250、500和1 000μmol/L硫酸镍处理6、12和24h。用酶联免疫吸附法检测培养上清液中T浓度,用实时荧光定量PCR技术检测间质细胞类固醇合成快速调节蛋白(StAR)、细胞色素P450胆固醇侧链裂解酶(P450scc)、3β-羟类固醇脱氢酶(3β-HSD)、类固醇17α-羟化酶(P450c17)mRNA的表达水平。结果人绒毛膜促性腺激素(hCG)和硫酸镍处理24h后,硫酸镍0μmol/L组与0h组比较,间质细胞分泌T浓度和StAR mRNA表达量均升高(P<0.05),而硫酸镍1 000μmol/L组T浓度和StAR mRNA表达量均降低(P<0.05)。NiSO4各浓度组与对照组比较,T浓度以及StAR、P450scc、3β-HSD和P450c17mRNA的表达量均明显降低(P<0.05)。结论硫酸镍引起的体外培养大鼠睾丸间质细胞T分泌量降低与类固醇合成酶基因表达下调有关。Objective To investigate the effects of niekel sulfate on the testosterone production and gene expression of steroidogenic enzymes of rat Leydig cells in vitro. Methods The rat Leydig cells purified by Percoil density gradient eentrifugation and differential adherent culture methods treated with 0, 250, 500 and 1000μmol/L nickel sulfate for 0, 6, 12 or 24h in vitro. The concentration of testosterone （T） in culture supernatant was determined by enzyme-linked immunosorbant assay. The mRNA expression levels of steroidogenie acute regulatory protein （STAR）, cytochrome P450 cholesteral side-chain cleavage enzyme （P450sec）, 3[3-hydroxysteroid dehydrogenase （3β-HSD）, steroid 17-alpha-Hydroxylase（P450c17） were measured by real-time PCR in Leydig cells. Results Compared with Oh group, the concentration of T and the level of StAR mRNA expression was significantly increased in 0 μmol/L nickel sulfate group（P〈0.05）, but the concentration of T and the mRNA expression level of StAR after treated with hCG for 24 h decreased in 1000 μmol/L nickel sulfate group （P〈0.05）. Compared with control group, the concentration of T and the mRNA expression levels of STAR, P450scc, 3β-HSD and P450c17 were decreased in all nickel sulfate exposure groups（P〈0. 05）. Conclusions The decrease of testosterone production induced by nickel sulfate has relation to the mRNA down-expression of steroidogenic enzymes in rat Leydig cells in vitro.

关 键 词：硫酸镍 间质细胞 睾酮 类固醇合成酶 

分 类 号：R114[医药卫生—卫生毒理学]