摘要
为了确定FSH和雌激素联合作用对培养条件下未成熟仔猪睾丸支持细胞中Cyclin D1mRNA和Cyclin E1mRNA表达的影响。以培养的仔猪睾丸支持细胞为研究材料,通过添加各种信号通路的抑制剂,应用实时荧光定量PCR检测Cyclin D1mRNA和Cyclin E1mRNA的相对表达量。FSH(50ng·mL-1)和17β-雌二醇(10-9 mol.L-1)联合作用时对Cyclin D1mRNA和Cyclin E1mRNA表达的影响与FSH或17β-雌二醇(10-9 mol.L-1)单独作用相比无显著影响(P>0.05);环磷酸腺苷抑制剂(Rp-cAMP)、L-Ca2+离子通道抑制剂(Verapamil)和ERK1/2抑制剂(U0126)单独作用时对Cyclin D1mRNA和Cyclin E1mRNA表达与空白对照组相比无显著影响(P>0.05),但以剂量依赖的方式抑制了FSH(50ng·mL-1)与17β-雌二醇(10-9 mol.L-1)联合作用对Cyclin D1mRNA和Cyc-lin E1mRNA的表达(P<0.05)。FSH和17β-雌二醇联合作用时通过cAMP、Ca2+内流和ERK1/2途径参与了FSH和17β-雌二醇对Cyclin D1mRNA和Cyclin E1mRNA在细胞中的表达。
The objective of the study was to identify whether the combination of FSH and 17-beta- estradiol regulated the mRNA expression of Cyclin D1 and Cyclin E1 in the cultured immature boar Sertoli cells. Cultured immature boar Sertoli cells were treated with the combination of 17- beta-estradiol （10-9 mol · L- 1） and FSH （50 ng · mL-1） and added a variety of different signa- ling pathway inhibitors. And Real-time PCR was applied to detect the mRNA expression of Cyc- lin D1 and Cyclin El. Compared to FSH （50 ng · mL-1） or 17-beta-estradiol （10-9 mol· L-1） alone, the combination of FSH and 17-beta-estradiol had no significant effect on the mRNA ex- pression of Cyclin D1 mRNA and Cyclin EI（P〉0.05）. In addition, Rp-cAMP （cAMP inhibi- tor）, Verapamil （L-type Ca2＋ ionic channel inhibitor） or U0126 （ERK1/2 inhibitor） alone had no significant effect on the mRNA expression of Cyclin D1 and Cyclin E1 in comparison to the con- trol group （no FSH orl7-beta-estradiol） （P〉0.05）. However, three inhibitors could reduce the mRNA expression of Cyclin D1 and Cyclin E1 in a dose-dependent way （P〈0.05, for all） whencompared to the combined FSH （50 ng · mL-1） and 17-beta-estradiol （10-9 mol · L- 1）. cAMP, Ca2＋ and ERK1/2 were involved in the effect of the combination of FSH and 17-beta-estradiol regulating the mRNA expression of Cyclin D1 and Cyclin El.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2013年第6期858-865,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金项目(31072183)
