L-半胱氨酸对绵羊卵巢颗粒细胞增殖、凋亡和类固醇激素分泌的影响

doi:10.11843/j.issn.0366-6964.2024.01.018

Abstract

Abstract: This study was conducted to investigate the effects of L-cysteine (L-Cys) on proliferation, apoptosis and the secretion of steroid hormones (progesterone and estrogen, P₄ and E₂) in ovine ovarian granulosa cells (GCs). Thirty sexually mature ewes aged between 1-1.5 years with the similar body weight and the same feeding condition were selected, and the fresh ovaries were collected. Then, GCs were collected from the dominant follicles (2-8 mm), and the GCs were randomly divided into 5 groups with 6 replicates per group. GCs were treated with 0, 100, 300, 500 and 700 μmol· L^-1 L-Cys for 48 h. The cell proliferation, apoptosis assay and the secretion of steroid hormones were measured using the CCK-8 kit, Annexin V-FITC/PI flow cytometry and ELISA, respectively. Real-time quantitative PCR and Western blot were used to analyze the mRNA and protein expression of the cell proliferation related genes (PCNA, CCND2, CCNB1), apoptosis-related genes (BAX, Caspase-3, Bcl-2) and steroid-secretion-related genes (STAR, 3β-HSD, CYP11A1, CYP19A1). The results showed that 100, 300 and 500 μmol·L^-1 L-Cys significantly promoted the cell viability (P<0.05) and significantly inhibited the apoptotic rate (P<0.05) of GCs. The addition of 300 μmol·L^-1 L-Cys significantly inhibited the secretion of P₄ (P<0.05). The mRNA expression of proliferation-related genes (PCNA, CCND2, CCNB1) were upregulated in the 300 μmol·L^-1 and 500 μmol·L^-1 L-Cys groups, while the protein expression of proliferation-related proteins (PCNA, CCND2, CCNB1) were increased in the 300 μmol·L^-1 L-Cys group (P<0.05). The mRNA and protein expressions of apoptosis-related genes (BAX, Caspase-3) were significantly downregulated in 100 μmol·L^-1 and 300 μmol·L^-1 L-Cys groups (P<0.05). The mRNA and protein expressions of the steroid-related genes (STAR, 3β-HSD) were significantly decreased in 300 μmol·L^-1 L-Cys group (P<0.05), but the mRNA and protein expressions of the steroid-related genes (CYP11A1, CYP19A1) were significantly increased in 100 μmol·L^-1 and 300 μmol·L^-1 L-Cys groups (P<0.05). In summary, L-Cys promoted cell proliferation and inhibited the apoptosis of ovine ovarian GCs by upregulating the mRNA and protein expressions of PCNA, CCNB1, CCND2, Bcl-2 and downregulating the mRNA and protein expressions of BAX, Caspase-3; L-Cys inhibited secretion of P₄ by downregulating the mRNA and protein expressions of STAR, 3β-HSD and upregulating the mRNA and protein expressions of CYP11A1.

Key words: sheep, ovarian granulosa cells, L-cysteine, proliferation, apoptosis, steroidogenesis

CLC Number:

S826.3

DUAN Xiangru, KANG Jia, YANG Ruochen, SHAN Xinyu, LI Taichun, ZHAO Wen, ZHANG Yingjie, LIU Yueqin. Effect of L-cysteine on Proliferation, Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 179-191.

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Effect of L-cysteine on Proliferation, Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells

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