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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OA北大核心CSTPCD

Effect of γ-aminobutyric Acid on Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells

中文摘要英文摘要

[目的]该文章旨在探究γ-氨基丁酸(GABA)对绵羊卵巢颗粒细胞(GCs)凋亡及类固醇激素分泌的影响.[方法]卵巢GCs培养液中分别添加 0、10-8、10-7、10-6 和 10-5 mol/L的GABA处理 24 h后,采用CCK-8 法和流式细胞术分别检测GCs增殖及凋亡情况,筛选出GABA适宜培养浓度用于后续试验,采用ELISA检测GABA对GCs雌激素(E2)及孕酮(P4)分泌的影响,RT-qPCR及Western blot技术检测促GCs增殖(PCNA、CCNB1、CCND1)、促凋亡(Caspase-3、Bax)、抗凋亡(Bcl-2)和类固醇激素合成相关基因(CYP11A1、StAR、3β-HSD、CYP19A1)mRNA及蛋白的表达.[结果]添加GABA处理绵羊卵巢GCs 24 h,各组均可显著促进GCs增殖(P<0.05),抑制其凋亡(P<0.05).GABA可显著上调PCNA、CCNB1、CCND1、Bcl-2 mRNA及蛋白表达量(P<0.05),显著下调Bax、Caspase-3 mRNA及蛋白表达量(P<0.05)和Bax/Bcl-2 比值(P<0.05).GABA可显著促进P4 的分泌(P<0.05),显著抑制E2 的分泌(P<0.05).显著上调StAR、3β-HSD、CYP11A1 的mRNA及蛋白表达水平(P<0.05),显著下调CYP19A1的mRNA及蛋白水平(P<0.05).[结论]GABA通过调节CCNB1、CCND1、PCNA、Caspase-3、Bcl-2、Bax基因表达促进GCs的增殖,抑制其凋亡;通过调节StAR、3β-HSD、CYP11A1、CYP19A1基因表达促进GCs的P4 分泌,抑制E2 生成.

[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.

单新雨;李太春;杨若晨;段香茹;康佳;张英杰;刘月琴

河北农业大学动物科技学院,保定 071000

γ-氨基丁酸绵羊颗粒细胞凋亡类固醇激素

γ-aminobutyric acidovine granulosa cellsapoptosissteroid hormone

《生物技术通报》 2024 (003)

312-321 / 10

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)

10.13560/j.cnki.biotech.bull.1985.2023-0784

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