膜联蛋白5对大鼠睾丸间质细胞中类固醇急性调节蛋白和睾酮合成相关酶表达的影响  

Effect of annexin 5 on StAR and testosterone synthesis related enzyme in rat Leydig cells

作　　者：陶晓倩[1] 林钗英[1] 柳海燕[1] 时姗姗[1] 路榕[1] 黄宇烽[1] 李晓军[1] 戈一峰[1] 崔英霞[1] 姚兵[1] 

机构地区：[1]江苏南京军区南京总医院解放军临床检验医学研究所,南京210002

出　　处：《北京大学学报（医学版）》2011年第2期183-188,共6页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30770801);全军"十一五"医药卫生科研基金(06H011);南京军区创新课题(07Z031)资助项目~~

摘　　要：目的:研究膜联蛋白5(annexin 5)对大鼠睾丸间质细胞(Leydig细胞)睾酮合成相关蛋白和酶表达的影响。方法:原代培养大鼠睾丸间质细胞24 h后,用10-9mol/L的annexin 5处理间质细胞12 h和24 h,分别提取细胞总RNA和总蛋白;反转录聚合酶链式反应(RT-PCR)检测类固醇急性调节蛋白(steroidogenic acute regulatory,StAR)、胆固醇侧链裂解酶(P450scc)、3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase,3β-HSD)、17α-羟化酶(17α-hydroxylase,CYP17A)、17β-羟基类固醇脱氢酶Ⅹ型(17β-HSD10)mRNA表达的改变,并用蛋白免疫印迹方法(Western blotting)检测蛋白表达水平的变化。结果:与对照组相比,在mRNA水平上,annexin 5处理细胞12 h后,只有17β-HSD10的表达升高了26%(P<0.05),其他差异均无统计学意义,而处理24 h后,StAR、P450scc和3β-HSD的表达分别升高55%、69%和59%(P<0.05),17β-HSD10升高了104%(P<0.01),17α-hydroxylase表达则无显著差异;在蛋白水平上,annexin 5处理细胞12 h后,17β-HSD的表达升高了39%(P<0.05),而处理24 h后发现,除StAR表达无显著变化外,P450scc、3β-HSD和17β-HSD的表达分别升高了35%、88%(P<0.05)和47%(P<0.01)。结论:Annexin 5对睾酮合成具有调节作用,这种作用是通过在基因水平和蛋白水平上影响P450scc、3β-HSD和17β-HSD的表达而实现的。Objective：To investigate the effect of annexin 5 on the expressions at mRNA levels and protein levels of StAR,P450scc,3β-HSD,17α-hydroxylase and 17β-HSD in rat Leydig cells.Method： The primary rat Leydig cells were cultured for 24 h and then stimulated with 10-9 mol/L annexin 5 for 12 h and 24 h respectively.Cellular total RNA and total protein were extracted respectively.The expressions of StAR,P450scc,3β-HSD,and 17α-hydroxylase and 17β-HSD10 mRNA were detected by reverse transcription-polymerase chain reaction（RT-PCR）and the protein levels were detected by Western blotting.Results： Compared with the control group,at the mRNA level,after being treated with annexin 5 for 12 h,only 17β-HSD10 expression had a 26% increase（P〈0.05） while the others had no significant dif-ference.The expressions of StAR,P450scc and 3β-HSD elevated 55%,69% and 59%（P〈0.05） respectively,and 17β-HSD10 increased 104%（P〈0.01） while 17α-hydroxylase had no significant difference after being treated with annexin 5 for 24 h.At the protein level,after being treated with annexin 5 for 12 h,17β-HSD expression had a 39% increase（P〈0.05）.After 24 h,P450scc,3β-HSD and 17β-HSD elevated 35%,88%（P〈0.05） and 47%（P〈0.01） respectively while StAR had no signifi-cant difference.Conclusion： Annexin 5 regulates testosterone synthesis by affecting the expressions of P450scc,3β-HSD and 17β-HSD at gene and protein levels.

关 键 词：膜联蛋白5 睾丸间质细胞 胆固醇侧链断裂酶 17-羟甾类脱氢酶类 

分 类 号：R345[医药卫生—基础医学]