T-2毒素对原代培养的小鼠睾丸间质细胞睾酮合成的影响

Effects of T-2 Toxin on Testosterone Biosynthesis in Primary Cultured Leydig Cells of Mice

目的探讨T-2毒素对原代培养的小鼠睾丸间质细胞(Leydig cells)睾酮合成的影响。方法建立健康清洁级成年雄性昆明小鼠睾丸间质细胞原代培养模型,使细胞悬液密度为5×105个/ml,采用改进3β-羟基类固醇脱氢酶(HSD)染色法进行纯度鉴定。将其分为空白对照[0ng/ml人绒毛膜促性腺激素(human chorionic gonadotrophin,hCG)+0mol/LT-2毒素]组、诱导对照(10ng/mlhCG+0mol/LT-2毒素)组、10ng/mlhCG+10-9mol/LT-2毒素组、10ng/mlhCG+10-8mol/LT-2毒素组、10ng/mlhCG+10-7mol/LT-2毒素组,培养24h后检测睾酮含量。结果新鲜分离的小鼠睾丸间质细胞在有血清培养液中培养24h后,细胞贴壁良好,其纯度在90%以上。在10ng/mlhCG诱导下,原代培养的小鼠睾丸Leydig细胞培养液中睾酮合成量明显增加,差异有统计学意义(P<0.05)。与诱导对照组相比,10ng/mlhCG+10-7、10-8、10-9mol/L的T-2毒素染毒组培养液中睾酮合成量均降低,差异有统计学意义(P<0.05或P<0.01);且睾酮合成量随着T-2毒素染毒浓度的升高而降低。结论T-2毒素可以直接降低原代培养的小鼠Leydig细胞睾酮合成功能。

Objective To investigate the effects of T-2 toxin on testosterone biosynthesis in primary cultured Leydig cell derived from the mouse testis. Methods Leydig cells isolated from Kunming male mice were adjusted to 5×10^5/ml and the purity was identified by the modified 3β-hydroxysteroid dehydrogenase （HSD） staining method. Blank control group （treated with 0 ng/ml hCG and 0 mol/L T-2 toxin） , inductive control group （treated with 10 ng/ml hCG and 0 mot/L T-2 toxin）, low dose T-2 toxin exposure group （treated with 10 ng/ml hCG and 10-9 mol/L T-2 toxin） , middle dose T-2 toxin exposure group （treated with 10 ng/ml hCG and 104 mol/L T-2 toxin） and high dose T-2 toxin exposure group （treated with 10 ng/ml hCG and 10-7 mol/L T-2 toxin） were designed,respectively. The testosterone level was measured after 24 h of incubation. Results After 24 hours culture in liquid medium contained serum, the fresh isolated Leydig cells grew well and the purity exceeded 90%. Through 10 ng/ml hCG induce, the testosterone level of Legdig cells increased significantly and the difference compared to blank control was of statistical sense （P〈0.05）. Compared to inductive control group,the testosterone level in Legdig cells decreased significantly in all T-2 toxin exposure groups with a dose dependant manner （P〈0.05 or P〈0.01）. Conclusion T-2 toxin can directly decrease the testosterone biosynthesis of the primary Leydig cells derived from the mouse testis.