丝裂原活化蛋白激酶信号传导通路对猪卵巢颗粒细胞分泌功能的调节作用  

Regulation of synthesis function of porcine ovarian granulose cells by mitogen-activated protein kinase signaling pathway

作　　者：邱学敏[1] 刘阳[1] 丛晶[1] 吴效科[1] 

机构地区：[1]黑龙江中医药大学附属第一医院妇产科,哈尔滨150040

出　　处：《中华妇产科杂志》2009年第12期933-935,共3页Chinese Journal of Obstetrics and Gynecology

基　　金：黑龙江省杰出青年基金（JC200804）

摘　　要：目的探讨丝裂原活化蛋白激酶（MAPK）信号传导通路对猪卵巢颗粒细胞分泌功能的调节作用。方法将体外培养的猪卵巢颗粒细胞分为两组，分别加入浓度为50μmo／／L的二甲基亚砜（DMSO）和MAPK抑制剂——PD98059（PD），作用48h后，将加入DMSO的细胞分为两组，观察1组细胞加入浓度为20μmol／L的腺苷酸环化酶激活剂，不加腺苷酸环化酶激活剂的为对照1组；将加入PD的细胞也分为两组，观察2组细胞加入浓度为20μmol／L的腺苷酸环化酶激活剂，不加腺苷酸环化酶激活剂的为对照2组，继续作用48h。以化学发光法测定4组细胞中睾酮、孕酮和雌二醇的水平；用RT-PCR技术检测17α羟化酶（CYP17）和细胞色素芳香酶P450的基因表达，以扩增产物的灰度比（GSR）表示。结果（1）生殖激素水平：对照1组睾酮、孕酮、雌二醇水平分别为（29．5±2．5）nmol／L、（80±5）nmol／L、（49±4）pmol／L，对照2组分别为（42．3±3．4）nmol／L、（170±15）nmol／L、（75±6）pmol／L，两组比较，差异有统计学意义（P〈0．05）；观察2组睾酮、孕酮、雌二醇水平分别为（106．2±7．6）nmol／L、（210±16）nmol／L、（130±11）pmol／L，观察1组分别为（47．2±3．5）nmol／L、（130±6）nmol／L、（81±6）pmol／L，两组比较，差异也有统计学意义（P〈0．05）；（2）酶表达变化：对照2组与对照1组比较，CYP17mRNA表达增强50％，而芳香酶P450mRNA表达下降20％，观察2组与观察1组比较，芳香酶P450、CYP17mRNA表达均增强，且以CYP17mRNA表达增强更明显，达到125％。结论MAPK信号传导通路对猪卵巢颗粒细胞的分泌功能具有抑制作用。Objective To investigate the role of mitogen-activated protein kinase（MAPK） signaling pathway regulating synthesis function of steroid hormone of porcine ovarian granulose cells. Methods Porcine ovarian granulose cells were cultured in medium added with dimethyl sulfoxide （DMSO） at concentration of 50 μmoL/L or with MAPK inhibitor PD98059 （PD） for 48 hours. Then, granulose cells in DMSO medium were added with activator of adenylate cyclase （20 μmol/L） or blank agent for next 48 hours incubation, which were defined as observation group 1 and control group 1, similarly, granulose cells in PD mediutn were also added with activator of adenylate cyclase （20 μmol/L） or blank agent for next 48 hours, which were defined as observation group 2 and control group 2. The level of estradiol （E2 ） , progesterone （P） and testosterone （T） were detected by chemoluminescence. The mRNA expression of 17alphahydroxylase/17,20-1yase （CYP17） and aromatase P450 （P450 atom）were assessed by RT-PCR. Results （ 1 ）Hormone expression： the level ofT, P and E2 were （29.5 ±2.5） nmol/L, （80 ±5） nmol/L, （49 ±4） pmol/L in control group 1 and （42.3 ± 3.4） nmol/L, （ 170 ± 15 ） nmol/L, （75 ± 6） pmol/L in control group 2, which showed significant difference （ P 〈 0. 05）. In the mean time, the level of T, P and E2 were （106.2 ±7.6） nmol/l,, （210 ± 16） nmol/L, （ 130 ± 11 ） pmol/L in observation group 2 and （47.2 ± 3.5） nmol/L, （130 ±6） nmol/L, （ 81 ±6） pmol/L in observation group 1, which also reach statistical difference （P 〈0. 05）. （2） The expression of enzyme： the expression of CYP17 mRNA was increased by 50% and P450 arom mRNA was decreased by 20% between control group 1 and 2. However, the mRNA expression of P450 arom and CYP17 were upregulated remarkably, especially, the expression of CYP17 mRNA were increased by 125%. Conclusion MAPK signaling pathway plays an inhibitory role in regulating synthesis of steroid hormone of o

关 键 词：卵巢 粒层细胞 丝裂原激活蛋白激酶类 类黄酮物质 福司柯林 

分 类 号：R711.75[医药卫生—妇产科学]