论文部分内容阅读
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 effect of nickel sulfate on the secretion of testosterone (T) and the gene expression of steroid synthase in cultured rat Leydig cells. Methods Rat Leydig cells were isolated, purified and cultured in vitro. The rats were treated with 0, 250, 500 and 1 000 μmol / L nickel sulfate for 6, 12 and 24 h, respectively. The concentration of T in culture supernatants was detected by enzyme-linked immunosorbent assay (ELISA). The levels of stAR, cytochrome P450 cholesterol side chain cleaving enzyme (P450scc), 3β-hydroxysteroid Dehydrogenase (3β-HSD), steroid 17α-hydroxylase (P450c17) mRNA expression levels. Results After treated with human chorionic gonadotropin (hCG) and nickel sulfate for 24 h, the levels of T and StAR mRNA in stromal cells were significantly increased in 0μmol / L nickel sulfate group compared with those in 0h group (P <0.05) The concentration of T and the expression of StAR mRNA in 1 000 μmol / L nickel group were decreased (P <0.05). Compared with the control group, the concentrations of T, and the expression of StAR, P450scc, 3β-HSD and P450c17 mRNA in NiSO4 groups were significantly decreased (P <0.05). Conclusion The decrease of T secretion of testicular stromal cells in vitro induced by nickel sulfate is related to the down regulation of steroid synthase gene expression.