畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (10): 1240-1245.

• 遗传繁育 • 上一篇    下一篇

FSH通过cAMP、Ca2+调节仔猪睾丸支持细胞的增殖

王鲜忠,周玉兰,赵伯川,周银涛,陈永军,张姣姣,王怡,张家骅*   

  1. 西南大学动物科技学院 重庆市牧草与草食家畜重点实验室,重庆 400716
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-10-25 发布日期:2010-10-25
  • 通讯作者: 张家骅

FSH Regulates Boar Sertoli Cell Proliferation via cAMP and Ca2+ Influx

WANG Xian-zhong, ZHOU Yu-lan, ZHAO Bo-chuan, ZHOU Yin-tao, CHEN Yong-jun, ZHANG Jiao-jiao, WANG Yi, ZHANG Jia-hua*   

  1. Chongqing Key Laboratory of Forage & Herbivorce, College of Animal Science and Technology, Southwest University,Chongqing 400716, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-10-25 Published:2010-10-25

摘要: 本研究旨在阐明FSH调节睾丸支持细胞增殖的机制。试验以2~3周龄的仔猪睾丸支持细胞为实验材料,采用体外培养模型,通过细胞数量检测、ELISA和Western blot等方法研究FSH对睾丸支持细胞的调节作用。试验结果如下:(1)在0~50 ng·mL-1内,随着FSH浓度的增加,睾丸支持细胞数量呈增加的趋势(P<0.05);当浓度为50 ng·mL-1时,FSH促增殖作用最明显;细胞内cAMP的浓度也随着FSH的浓度而增加(P<0.05);(2)FSH作用后5 min内就激活了ERK1/2级联反应,在48 h内,ERK1/2的激活有一定的周期性; 加入ERK1/2的抑制剂PD98059和U0126明显的降低了FSH诱导的睾丸支持细胞的增殖和PCNA的表达(P<0.05);(3)加入FSH和Forskolin增强了细胞的增殖和ERK1/2的激活(P<0.05);而加入Rp-cAMP则明显降低了FSH诱导的细胞增殖和ERK1/2的激活(P<0.05); L-Ca2+通道抑制剂Verapamil抑制了细胞的增殖和ERK1/2的激活;Rp-cAMP与Verapamil共同作用强于单独作用,表现出一定的协同效应(P<0.05)。结果表明,FSH通过cAMP、Ca2+激活ERK1/2,并通过ERK1/2调节PCNA的表达进而调节支持细胞的增殖。

关键词: 促卵泡素, 细胞增殖, 睾丸支持细胞, 细胞外调节的蛋白激酶1/2

Abstract: This experiment was conducted to study mechanisms of FSH regulating Sertoli cell proliferation. Sertoli cells of 2-3 week-old piglet were used as cell models, and cell viability, ELSIA and Western blot were used to explain the effect of FSH. The results were as follows: (1) From 0 to 50 ng·mL-1, with the increase of the concentration of FSH, Sertoli cell number showed an upward trend(P<0.05); at 50 ng·mL-1, effect of FSH on Sertoli cell proliferation was most significant; the concentration of intracellular cAMP also increased, following the increase of the concentration of FSH(P<0.05); (2) FSH begun to activate ERK1/2 cascade reaction 5 min after its affecting and the activating model had a periodicity; ERK1/2 inhibitor (PD98059 and U0126) could significantly inhibit the FSH-induced Sertoli cell proliferation and the expression of PCNA (proliferating cell nuclear antigen) (P<0.05); (3) Both of FSH and Forskolin could enhance cell proliferation and ERK1/2 activation (P<0.05), while Rp-cAMP and L-Ca2+ channel inhibitor (Verapamil) significantly both inhibited the FSHinduced cell proliferation and ERK1/2 activation(P<0.05); Rp-cAMP and Verapamil showed synergism effect (P<0.05). These results indicated that FSH could activate ERK1/2 via cAMP and Ca2+,which increased the expression of PCNA, and promoted Sertoli cell proliferation.

Key words: FSH, cell proliferation, Sertoli cell, ERK1/2