GnRH抑制剂对大鼠睾丸间质细胞annexin 5和3β-HSD蛋白表达水平的影响

Effects of GnRH-antagonists on the protein expression of annexin 5 and 3β-HSD in rat Leydig cell

目的探讨西曲瑞克(GnRHant)对大鼠睾丸间质细胞中膜联蛋白Ⅴ(annexin5)和3 β-羟类固醇脱氢酶(3 β-HSD)表达的影响。方法体外分离、纯化和培养大鼠睾丸间质细胞原代培养,采用3β-HSD免疫细胞化学方法对间质细胞进行鉴定,用不同终浓度的GnRHant(分别为10^(-6)、10^(-7)和10^(-8)mol/L)作用于间质细胞24h后,通过Western Blot检测间质细胞中annexin5和3 β-HSD的表达情况。结果间质细胞经GnRHant处理后,Western Blot结果显示,当GnRHant的终浓度为10^(-6)mol/L和10^(-7)mol/L时,实验组大鼠睾丸间质细胞annexin 5的表达量都显著地降低,分别降低了62.35%和40.82%,两者比较,差异具有统计学意义(P<0.05);当GnRHant的终浓度为1 0^(-6)mol/L和10^(-7)mol/L时,实验组大鼠睾丸间质细胞3β-HSD的表达量也降低,分别降低了53.33%和23.85%(P分别<0.01和<0.05),当GnRHant的终浓度为10^(-8)mol/L时,与对照组比较,差异均没有统计学意义(P>0.05)。结论 GnRH抑制剂在原代培养的睾丸间质细胞中,对annexin5和3 β-HSD的表达具有一定调控作用。

Objective To investigate the effects of GnRH-antagonists on the protein expression of annexin5 and 3β-HSD in Leydig cells. Methods The primary Leydig cells were isolated, purified and cultured in vitro. The cells were identified by 3β-HSD expression. Leydig cells were treated by different concentrations of GnRHant for 24 hour. The expressions of annexin5 and 3β-HSD were detected by Western blot. Results The protein expression of annexin5 and 3β-HSD decreased in the treated group with GnRHant compared with that of control groups. Under treated by the GnRH-antagonists with 10^6mol/L and 10-Tmol/L, the expression of annexin5 in Leydig cells decreased by 62.35% （ P〈0.05, P=0.047） and 40.82%（P〈0.05, P=0.04） respectively, and the expression of 3β-HSD decreased by 53.33% （P〈0.01, P=0.01）and 23.85%（P〈0.05, P=0.037） respectively. There has no significant change in group GnRH-antagonists 10^-8mol/L. Conclusion GnRH-antagonists might regulate the expression of annexin5 and 3β-HSD in primary Leydig cells.