摘要
目的探讨锰对体外培养大鼠Leydig细胞P450scc的影响。方法建立体外培养大鼠Leydig细胞的原代培养方法,染锰(0、10、30和100μmol/L)24 h后,原子吸收光谱法检测细胞内外锰的含量,ELISA方法检测细胞培养上清液中睾酮及细胞内外P450scc的含量,并运用反转录-聚合酶链反应(RT-PCR)的方法检测Leydig细胞P450sccmRNA的表达水平。结果与对照组相比,在设计剂量范围内,随着染锰剂量加大,细胞培养上清液中睾酮及细胞内Mn的含量逐渐增加,30和100μmol/L时,有统计学意义(P<0.05)。细胞内外P450scc含量不断增加,30和100μmol/L时,细胞内该酶的增加有统计学意义(P<0.05);10、30和100μmol/L时,细胞外该酶的增加有统计学意义,P<0.05。P450scc mRNA的表达水平升高,100μmol/L时,有统计学意义,P<0.05。结论在该试验条件下,MnCl2可上调P450scc基因的表达,增加细胞内P450scc含量,促进体外培养Leydig细胞合成睾酮。
Objective To investigate the effect of manganese on P450scc in rat Leydig cells.Methods After establishment of primary culture method of rat Leydig cells,different concentrations of MnCl2（0,10,30,100 μmol /L） were added into cultured cell medium,24 hours later,the contents of Mn in cell culture supernatant and in Leydig cells were detected by atomic absorption spectrometry（AAS）,The contents of testosterone,P450scc in cell culture supernatant and P450scc in Leydig cells were measured by ELISA,The expression of P450scc mRNA was quantified by RT-PCR.Results Compared with the control group,The contents of Mn in Leydig cells and testosterone in cell culture supernatant increased gradually,at the doses of 30,100 μmol /L,the differences were significant（P〈0.05）.The content of P450scc increased gradually,at the doses of 30,100 μmol /L,the differences were significant in Leydig cells（P〈0.05）;and at the doses of 10,30,100 μmol /L,the differences were significant in cell culture supernatant（P〈0.05）.The expression of P450scc mRNA increased and the difference was significant at the dose of 100 μmol /L（P〈0.05）.Conclusions MnCl2 could upregulate the expression of P450Scc gene,increase the content of P450Scc,and promote the production of testosterone of Leydig cells in vitro.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2010年第4期279-281,共3页
Journal of Toxicology
基金
国家自然科学基金(30760209)
