GnIH对小鼠卵巢颗粒细胞增殖、凋亡和雌激素分泌的影响

doi:10.11843/j.issn.0366-6964.2025.05.022

Abstract

Abstract:

The experiment aimed to investigate the effects of interfering and overexpressing GnIH on the proliferation, apoptosis and estrogen secretion in mouse ovarian granulosa cells. In this study, healthy female ICR mouse of 21-23 days old were selected. GCs were isolated from the mouse ovaries and identified using follicle-stimulating hormone receptor (FSHR). qRT-PCR was used to detect the expression of GnIH and GnIH-R genes in mouse granulosa cells. pshRNA-1, 2, 3, 4 interfering plasmid vectors and pLV-GnIH overexpressed plasmid vectors were constructed to overexpress and interfere with GnIH gene in mouse granulosa cells, respectively (GnIH interference groups were divided into pshNC group and pshRNA1-4 groups, and GnIH overexpression groups were divided into PLV-NC group and PLV-GnIH group). The interference and overexpression efficiency were detected by lentivirus titer detection and RT-qPCR, and the effects of psh-RNA and pLV-RNA on the proliferation and apoptosis of granulosa cells were detected by MTT and Annexin V-FITC/PI flow cytometry. The concentrations of progesterone and estradiol were detected by ELISA, all the above results were repeated 3 times. The results showed that pshRNA-1, 2, 3, 4 significantly interfered with GnIH expression at 48 h and 96 h (P < 0.01), but only pshRNA-4 significantly decreased GnIH expression at 72 h (P < 0.01). At 72 h, pLV-GnIH significantly up-regulated the expression of GnIH (P < 0.001). Overexpression of GnIH significantly down-regulated the proliferation activity of granulosa cells and promoted cell apoptosis (P < 0.001), and significantly inhibited granulosa cell estrogen secretion (P < 0.001). After transfection with pshRNA-2, 4 for 72 and 96 h, the proliferative activity of granulosa cells was extremely significantly increased (P < 0.01), the secretion of estrogen was significantly enhanced (P < 0.05), and apoptosis was significantly decreased(P < 0.05 and P < 0.01). In conclusion, inhibition of GnIH expression can promote the proliferation of granulosa cells and the secretion of estrogen, and down-regulate cell apoptosis, while promotion of GnIH expression can inhibit the proliferation of granulosa cells and the secretion of estrogen, and promote cell apoptosis.

Key words: GnIH, granulosa cells, proliferation, apoptosis, estrogen secretion

CLC Number:

S814.1

GAO Zhengjie, LUO Ping, LI Bocheng, WANG Shuilian. Effects of GnIH on Proliferation, Apoptosis and Estrogen Secretion of Mouse Ovarian Granulosa Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2230-2242.

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基因 Gene	引物序列 Primer sequence	产物大小/bp Product length
musGnIH	F: ATGCCTATCACATTGCTG R: CAAGGTGAAGACGGAAGC	114
musGnIH-R	F: CAGAAAGCAGGTGGATGG R: CTCCAGGTCCCCAAATCC	247
musGAPDH	F: CCGTGTTCCTACCCCCAATG R: AGCCCAAGATGCCCTTCAGT	128

序列名称 Sequence name	序列信息 Sequence information
GnIH-siRNA1	GCAACTTCAAGCTTCTTAACA
GnIH-siRNA2	GCAGGGACCAGGAGCCATTTC
GnIH-siRNA3	GCTCCAGCGAGTTGCTCTACG
GnIH-siRNA4	GCGAGTTGCTCTACGTCATGA

基因 Gene	引物序列 Primer sequence	产物大小/bp Product length
musGnIH	F: CTGGCTAGGATGGCGAATGG R: GCTCATGGTGGACCTTCCTA	111
musGAPDH	F: CCAGGTGGTCTCCTCTGA R: GCTGTAGCCAAATCGTTGT	127

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Effects of GnIH on Proliferation, Apoptosis and Estrogen Secretion of Mouse Ovarian Granulosa Cells

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