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目的探讨锰对体外培养Leydig细胞3β-HSD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3β-HSD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3β-HSD mRNA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、100μmol/L时,有统计学意义(P<0.05)。细胞内外3β-HSD的含量及3β-HSD mRNA的表达水平均在染锰10μmol/L时升高,之后不断降低,100μmol/L时,有统计学意义(P<0.05)。结论在该实验条件下,MnCl2可以改变3β-HSD基因表达及蛋白合成,影响体外培养Leydig细胞合成睾酮。
Objective To investigate the effect of manganese on 3β-HSD in cultured Leydig cells. Methods Primary culture method of rat Leydig cells cultured in vitro was established. After manganese (0, 10, 30 and 100 μmol / L) was dipped for 24 h, the content of manganese in cells was detected by atomic absorption spectrometry. The cell culture supernatants 3β-HSD in Leydig cells were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Compared with the control group, the testosterone and intracellular Mn contents in the cell culture supernatant gradually increased with the increase of manganese dose within the range of the designed dose (P <0.05), P <0.05). The content of 3β-HSD and the expression of 3β-HSD mRNA increased both in and out of 10μmol / L manganese, and then decreased continuously (100μmol / L, P <0.05). Conclusion Under the experimental conditions, MnCl2 can change the gene expression and protein synthesis of 3β-HSD, and affect the synthesis of testosterone in Leydig cells cultured in vitro.