Niu Chunyang, Zhang Sujuan, Mo Guilin, Jiang Yilong, Li Liang, Xu Hengyong, Han Chunchun, Zhao Hua, Yan Yanhong, Hu Shenqiang, Hu Jiwei, Kang Bo, Jiang Dongmei
College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, P. R. China.
Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, P. R. China.
Poult Sci. 2021 Aug;100(8):101226. doi: 10.1016/j.psj.2021.101226. Epub 2021 Apr 28.
Ornithine decarboxylase (ODC) plays an indispensable role in the process of polyamine biosynthesis. Polyamines are a pivotal part of living cells and have diverse roles in the regulation of cell proliferation and apoptosis, aging and reproduction. However, to date, there have been no reports about ODC regulating follicular development in goose ovaries. Here, we constructed ODC siRNA and overexpression plasmids and transfected them into goose primary granulosa cells (GCs) to elucidate the effects of ODC interference and overexpression on the polyamine metabolism, hormone levels, cell apoptosis and proliferation of granulosa cells. After interfering with ODC in GCs, the mRNA and protein levels of ODC and the content of putrescine were greatly decreased (P < 0.05). When ODC was overexpressed, ODC mRNA and protein levels and putrescine content were greatly increased (P < 0.05). The polyamine-metabolizing enzyme genes ornithine decarboxylase antizyme 1 (OAZ1) and spermidine / spermine-N1-acetyltransferase (SSAT) were significantly increased, and spermidine synthase (SPDS) was significantly decreased when ODC was downregulated (P < 0.05). OAZ1, SPDS and SSAT were significantly increased when ODC was upregulated (P < 0.05). In addition, after interference with ODC, progesterone (P4) levels in the culture medium of GCs increased greatly (P < 0.05), while the overexpression of ODC caused the P4 level to decrease significantly (P < 0.05). After ODC downregulation, granulosa cell activity was significantly reduced, the apoptosis rate was significantly increased, and the BCL-2 / BAX ratio was downregulated (P < 0.05). Under ODC overexpression, the activity of GCs was notably increased, the apoptosis rate was significantly reduced, and the BCL-2 / BAX protein ratio was upregulated (P < 0.05). Our study successfully induced ODC interference and overexpression in goose ovarian GCs, and ODC regulated mainly putrescine content in GCs with a slight influence on spermidine and spermine. Moreover, ODC participated in the adjustment of P4 levels in the culture medium of GCs, promoted granulosa cell proliferation and inhibited granulosa cell apoptosis.
鸟氨酸脱羧酶（ODC）在多胺生物合成过程中发挥着不可或缺的作用。多胺是活细胞的关键组成部分，在细胞增殖与凋亡、衰老和繁殖的调节中具有多种作用。然而，迄今为止，尚无关于ODC调节鹅卵巢卵泡发育的报道。在此，我们构建了ODC小干扰RNA（siRNA）和过表达质粒，并将它们转染到鹅原代颗粒细胞（GCs）中，以阐明ODC干扰和过表达对颗粒细胞多胺代谢、激素水平、细胞凋亡和增殖的影响。在GCs中干扰ODC后，ODC的mRNA和蛋白水平以及腐胺含量大幅下降（P < 0.05）。当ODC过表达时，ODC的mRNA和蛋白水平以及腐胺含量大幅增加（P < 0.05）。当ODC下调时，多胺代谢酶基因鸟氨酸脱羧酶抗酶1（OAZ1）和亚精胺/精胺-N1-乙酰转移酶（SSAT）显著增加，而亚精胺合成酶（SPDS）显著下降（P < 0.05）。当ODC上调时，OAZ1、SPDS和SSAT显著增加（P < 0.05）。此外，干扰ODC后，GCs培养基中的孕酮（P4）水平大幅升高（P < 0.05），而ODC的过表达导致P4水平显著降低（P < 0.05）。ODC下调后，颗粒细胞活性显著降低，凋亡率显著增加，BCL-2/BAX比值下调（P < 0.05）。在ODC过表达情况下，GCs的活性显著增加，凋亡率显著降低，BCL-2/BAX蛋白比值上调（P < 0.05）。我们的研究成功在鹅卵巢GCs中诱导了ODC干扰和过表达，并且ODC主要调节GCs中的腐胺含量，对亚精胺和精胺影响较小。此外，ODC参与了GCs培养基中P4水平的调节，促进颗粒细胞增殖并抑制颗粒细胞凋亡。
