孕酮在颗粒细胞中通过cAMP/PKA信号通路影响卵母细胞发育  

Progesterone Affects Oocyte Development Through cAMP/PKA Signaling Pathway in Granulosa Cells

作　　者：沈亚楠 季贵义 王进芳 胡蓉[3] SHEN Ya’nan;JI Guiyi;WANG Jinfang;HU Rong(CollegeofClinicalMedicine,Ningxia Medical University,Yinchuan 750004,China;Department of Obstetric,the General Hospital of Ningxia Medical University,Yinchuan 750004,China;Reproductive Medicine Center,the General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]宁夏医科大学总医院产科,银川750004 [3]宁夏医科大学总医院生殖医学中心,银川750004

出　　处：《宁夏医科大学学报》2023年第6期600-604,611,共6页Journal of Ningxia Medical University

基　　金：宁夏回族自治区卫生健康系统科研课题项目(2022-NWKY-058)。

摘　　要：目的探究孕酮在颗粒细胞中影响卵母细胞发育的分子机制。方法将人卵巢颗粒细胞(KGN)用不同浓度孕酮(P4)(0、1000、2000、3000 ng·mL^(-1))处理之后,RT-PCR和Western blot法检测不同浓度P4处理后卵泡刺激素受体(FSHR)和干细胞因子(SCF)的表达情况;用rFSH、H-89、forskolin、forskolin+H-89、P4+H-89和P4分别处理KGN细胞后,RT-PCR和Western blot法检测SCF的基因及蛋白表达情况;CCK-8法检测不同浓度P4处理后的细胞存活率;Western blot法检测不同浓度P4处理后Caspase3、Bcl-2、Bax相关凋亡蛋白的表达情况。结果用不同浓度P4(0、1000、2000、3000 ng·mL^(-1))处理之后,FSHR和SCF的基因和蛋白的相对表达量均下降(P均<0.05)。用rFSH、H-89、forskolin、forskolin+H-89、P4+H-89和P4分别处理KGN细胞后,显示P4可以下调forskolin诱导的SCF的表达。P4抑制颗粒细胞系KGN细胞的存活率,并促进Caspase3、Bcl-2、Bax相关凋亡蛋白的表达。结论P4通过cAMP/PKA通路影响颗粒细胞中SCF的表达,并且抑制细胞的存活,促进细胞凋亡。Objective To explore the molecular mechanism of progesterone affecting oocyte development in granulosa cells.Methods Human ovarian granulosa cell line(KGN)were treated with different concentrations of P4(0,1000,2000,3000 ng·mL^(-1))expression of follicle stimulating hormone receptor(FSHR)and stem cell factor(SCF)was detected by RT-PCR and Western blot.KGN cells were treated with rFSH,H-89,forskolin,forskolin+H-89,P4+H-89 and P4,respectively.The expression of SCF mRNA and protein was detected by RT-PCR and Western blot.The cell viability treated with different concentrations of P4 was detected by CCK8 assay.Western blot was used to detect the expression of Caspase3,Bcl-2,and Bax-related apoptotic proteins after different concentrations of P4 treatment.Results The relative expression levels of FSHR and SCF genes and proteins decreased after treatment with different concentrations of P4(0,1000,2000,3000 ng·mL^(-1))(P all<0.05).After KGN cells were treated with rFSH,H-89,forskolin,forskolin+H-89,P4+H-89 and P4,respectively,P4 down-regulated forskolin-induced SCF expression.P4 inhibited the cell viability of KGN cells and promoted the expression of Caspase3,Bcl-2,and Bax-related apoptotic proteins.Conclusion P4 affects the expression of SCF in granulosa cells through cAMP/PKA pathway,inhibits cell survival and promotes apoptosis.

关 键 词：孕酮 卵泡刺激素受体 卵母细胞 

分 类 号：R339.2[医药卫生—人体生理学]