DBP介导Cx43对大鼠睾丸Leydig细胞睾酮合成的影响

Effects of Cx43 Mediated by DBP on the Production of Testosterone in Testicle Leydig Cell in Rats

目的观察邻苯二甲酸二丁酯(DBP)对成年大鼠睾丸Leydig细胞连接蛋白43(Cx43)及其引起睾酮分泌变化的影响,探讨DBP对睾酮(T)的调节是否与Cx43有关.方法将原代培养纯化的leydig细胞分为3组:50μg/mL DBP处理组,50μg/mL DBP+10μmol/L前列腺素E2(PGE2)组及DMSO处理组,分别同时处理细胞24h;采用细胞免疫荧光法检测细胞膜Cx43表达变化,Western印记法观察Cx43总蛋白量的变化,应用睾酮试剂盒测定睾酮值.结果与对照组相比,DBP处理24h后,Cx43表达与睾酮值均降低,差异具有统计学意义(P<0.05);但加入Cx43增强剂(PGE2)组与DBP组相比,CX43表达明显恢复,但T值变化不明显(P>0.05).结论DBP对睾酮及Cx43均有影响,但对Leydig细胞睾酮合成抑制作用并不一定是通过调节Cx43的表达来实现.

Objective To observe the effects of DBP on Cx43 in testicle Leydig cell and testosterone（T）secretion in adult rats,and to explore the relationship between Cx43 and testosterone secretion regulated by DBP.Method Established the primary culture system,Leydig cells were incubated in 50 μg / m L DBP,50μg/mL DBP ＋ 10μmol/L PGE2 and DMSO（control group）.After culturing for 24 h,the expression of Cx43 on cell membrane was investigated with immunofluorescence and the total protein of Cx43 was evaluated by Western blotting.The production of testosterone was measured by testosterone ELISA Kit.Results Comparing with the control group,the expression of Cx43 and testosterone production decreased after incubated by DBP for 24 hours（P〈0.05）.But in the group of DBP ＋ PGE2,the production of T didn＇t recover with the up-regulation of Cx43（P 〉0.05）.Conclusion Both Cx43 and testosterone can be regulated by DBP,but its inhibitory effect on the synthesis of T in Leydig cells may be not necessarily achieved through the regulation on the expression of Cx43 necessarily.