生物技术通报2024，Vol.40Issue(3)：312-321,10.DOI:10.13560/j.cnki.biotech.bull.1985.2023-0784
γ-氨基丁酸对绵羊卵巢颗粒细胞凋亡及类固醇激素分泌的影响
Effect of γ-aminobutyric Acid on Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells
摘要
Abstract
[Objective]This study was conducted to investigate the effects of γ-aminobutyric acid(GABA)on the apoptosis and the secretion of steroid hormones in ovine ovarian granulosa cells(GCs).[Method]The GCs were treated with 0,10-8,10-7,10-6 and 10-5 mol/L GABA for 24 h,respectively.Then,the cell proliferation and apoptosis were detected by CCK-8 and flow cytometry.And the appropriate culture concentration of GABA was screened for subsequent experiments.ELISA was used to detect the secretion levels of estrogen(E2)and progesterone(P4)in the culture supernatant after GCs were treated with the appropriate concentration.The qRT-PCR and Western blot were to detect the mRNA and protein expressions of GCs proliferation-promoting genes(PCNA,CCNB1,and CCND1),anti-apoptosis genes(Bcl-2),apoptosis-promoting genes(Bax,Caspase-3)and steroid hormone synthesis-related genes(StAR,3β-HSD,CYP11A1,CYP19A1).[Result]The 10-6 mol/L GABA treatment for 24 h significantly promoted GCs proliferation(P<0.05)and significantly inhibited its apoptosis(P<0.05).The mRNA and protein expressions of PCNA,CCNB1,CCND1 and Bcl-2 in the experimental group were higher than those in the control group(P<0.05).Compared with the control group,the mRNA and protein expressions of Bax and Caspase-3 in the experimental group decreased significantly(P<0.05),as well as Bax/Bcl-2 decreased significantly(P<0.05).Treatment of ovine ovarian GCs with 10-6 mol/L GABA for 24 h significantly promoted the synthesis of P4(P<0.05)and significantly inhibited the synthesis of E2(P<0.05).Compared with the control group,the mRNA and protein expressions of StAR,3β-HSD and CYP11A1 in the experimental group increased significantly(P<0.05)and the mRNA and protein expressions of CYP19A1 decreased significantly(P<0.05).[Conclusion]The results demonstrate that GABA treatment could promote GCs proliferation as well as inhibit GCs apoptosis by regulating the expression of CCNB1,CCND1,PCNA,Caspase-3,Bax and Bcl-2,and promote the synthesis of P4 as well as inhibit the synthesis of E2 by regulating the expressions of StAR,3β-HSD,CYP11A1 and CYP19A1.关键词
γ-氨基丁酸/绵羊颗粒细胞/凋亡/类固醇激素Key words
γ-aminobutyric acid/ovine granulosa cells/apoptosis/steroid hormone引用本文复制引用
单新雨,李太春,杨若晨,段香茹,康佳,张英杰,刘月琴..γ-氨基丁酸对绵羊卵巢颗粒细胞凋亡及类固醇激素分泌的影响[J].生物技术通报,2024,40(3):312-321,10.基金项目
国家肉羊产业技术体系(CARS-38),国家绒毛用羊产业技术体系(CARS-39) （CARS-38）
