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

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5×10~3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0 ng/ml人绒毛膜促性腺激素(human chorionic gonadotrophin,hCG)+0 mol/L T-2毒素]组、诱导对照(10 ng/ml hCG+0 mol/L T-2毒素)组、10 ng/ml hCG+10~(-9)mol/L T-2毒素组、10 ng/ml hCG+10~(-8)mol/T~(-2)毒素组、10 ng/ml hCG+10~(-7) mol/L T-2毒素组,培养24 h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/ml hCG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,10 ng/ml hCG+10~(-7)、10~(-8)、10~(-9)mol/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.

Abstract：

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 n/ml hCG and 0 mol/L T-2 toxin),inductive control group(treated with 10 ng/ml hCG and 0 mol/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 10~(-8) 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.

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

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 n/ml hCG and 0 mol/L T-2 toxin),inductive control group(treated with 10 ng/ml hCG and 0 mol/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 10~(-8) 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.