GnRH类似物对大鼠睾丸间质细胞MAPK的影响

目的:研究大鼠睾丸间质细胞中GnRH激动剂(GnRHa)和拮抗剂(GnRHant)对促分裂原活化蛋白激酶(MAPK)途径的影响。方法:原代培养大鼠睾丸间质细胞24h后,血清饥饿2h。GnRHa(10-7mol/L)或Gn-RHant(10-6mol/L)处理大鼠睾丸间质细胞不同时间(0、5、15、30、60、90min)后,Western印迹检测磷酸化ERK(p-ERK)和磷酸化p38(p-p38)蛋白水平以0min组为对照。此外,不同终浓度(1、5、10、20μmol/L)的PKC抑制剂GF109203X和GnRHa或GnRHant共同作用后,Western印迹检测p-ERK蛋白水平。结果:GnRHa或GnRHant刺激间质细胞不同时间后,p-p38蛋白水平与对照组比较均无显著性差异(P>0.05);加入不同终浓度的PKC抑制剂GF109203X处理细胞20min后,再用GnRHa刺激细胞10min,发现GF109203X在终浓度为10μmol/L和20μmol/L时p-ERK水平显著下降(P<0.05)。GnRHant刺激间质细胞后,p-ERK水平在15min时升高了约65%(P<0.05);30min时升高了约81%,达到最高水平(P<0.05);60min时开始下降,90min时恢复到基础水平。不同浓度GF109203X处理细胞20min后,再用GnRHant刺激细胞30min,p-ERK水平与对照组比较无显著性差异(P>0.05)。结论:GnRHa可能通过PKC途径来诱导ERKMAPK信号通路的活化,而GnRHant对ERKMAPK信号通路的激活作用并非通过PKC途径来实现的;p38可能不参与间质细胞中GnRH类似物作用的MAPK分子信号通路。

Effects of GnRH analogues on MAPK pathway in rat Leydig cells

Objective:To investigate the effects of GnRH analogues GnRHa and GnRHant on the MAPK pathway in rat Leydig cells.Methods:Rat Leydig cells were primarily cultured for 24 hours in vitro and serum-starved for 2 hours,followed by treatment with GnRHa(10-7 mol/L)or GnRHant(10-6 mol/L)for 0,5,15,30,60 and 90 minutes,with the 0 min group as the control.Then the protein levels of phosphorylated ERK(p-ERK)and phosphorylated p38(p-p38)were detected by Western blot,and that of p-ERK determined by the same means after ...