FGF21增强线粒体功能抑制猪卵巢颗粒细胞凋亡

doi:10.11843/j.issn.0366-6964.2023.03.017

Abstract

Abstract: The purpose of this study was to investigate the effect of fibroblast growth factor 21 (FGF21) on the apoptosis of porcine (Sus scrofa) ovarian granulosa cells and its regulatory mechanism. In this study, follicular fluid was collected from fresh ovarian tissue of Large white×Landrace sows ((8±1) months old) to separate granulosa cells as the research object. Granulosa cells were treated by RNAi technology and added with different concentrations of FGF21 recombinant protein (0, 0.1, 1, 10 ng·mL^-1) for 24 h. After 24 h, the cell apoptosis was detected by flow cytometry, and the expression levels of BAX and BCL2 in granulosa cells were investigated by Western blot, qRT-PCR. In order to further clarify the relationship between the effect of FGF21 on apoptosis of granulosa cells and mitochondria, the protein levels of mitochondrial complexes in granulosa cells after different treatments, the number of mitochondria in cells, ATP concentration, total antioxidant activity, reactive oxygen species levels and the expression levels of mitochondrial production-related genes were detected. The study results showed that after knockdown of FGF21 expression in granulosa cells by RNAi technology, the proportion of cells in late apoptosis was significantly increased (P<0.05) and the proportion of viable cells was significantly decreased (P<0.05). At the same time, the mRNA and protein levels of the pro-apoptotic gene BAX in granulosa cells in the FGF21 knockdown group were significantly increased (P<0.05), while the expression of the apoptosis-inhibiting gene BCL2 was significantly decreased (P<0.05). On the contrary, the addition of recombinant protein FGF21 effectively prevented the apoptosis of granulosa cells, and up-regulated the expression of anti-apoptotic gene BCL2 (P<0.05) and inhibited the expression of pro-apoptotic gene BAX (P<0.05). Further study found that after knockdown of FGF21 expression in granulosa cells, the expression of mitochondrial oxidative phosphorylation complex II succinate dehydrogenase complex iron sulfur subunit B (SDHB) was significantly decreased (P<0.05). At the same time, the copy number of mitochondrial DNA was significantly decreased (P<0.001); In addition, it was found that, after FGF21 knockdown, the concentration of adenosine triphosphate (ATP) and the total antioxidant capacity (T-AOC) significantly decreased (P<0.001), the expression of mitochondrial production-related genes was significantly inhibited (P<0.05), and the level of cellular reactive oxygen species (ROS) was significantly increased (P<0.05). In contrast, FGF21 recombinant protein treatment significantly increased SDHB expression (P<0.01) and mitochondrial number (P<0.05), and significantly increased mitochondrial DNA copy number (P<0.01) and ATP concentration (P<0.05), T-AOC activity (P<0.05) and expression of mitochondrial production-related genes, while ROS levels were significantly decreased (P<0.05). The above results indicated that FGF21 promoted mitochondrial biosynthesis in granulosa cells and maintained the balance of intracellular oxidation-antioxidant system, thereby inhibiting the occurrence of apoptosis of granulosa cells. This study provides a theoretical basis for further revealing the potential regulatory mechanism of follicular development in livestock.

Key words: ovarian granulosa cells, FGF21, apoptosis, mitochondrial function, sows

CLC Number:

S828.3

HU Yamei, SONG Xiangrong, HUANG Liang, ZHANG Lutong, GAO Lei, PANG Weijun, YANG Gongshe, CHU Guiyan. FGF21 Enhances Mitochondrial Function and Inhibits Apoptosis of Porcine Ovarian Granulosa Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1034-1045.

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FGF21 Enhances Mitochondrial Function and Inhibits Apoptosis of Porcine Ovarian Granulosa Cells

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