Bta-miR-101对牛睾丸支持细胞增殖、凋亡及分泌的影响

doi:10.11843/j.issn.0366-6964.2024.03.017

Abstract

Abstract: The study aimed to explore the effects of bta-miR-101 on bovine testicular sertoli cells' proliferation, apoptosis and secretion.In this study, testicular tissues of healthy bulls (n=3) aged 3 days and 13 months were collected to construct the expression profile of Angus cattle tissues. The differences in bta-miR-101 expression were verified between the two periods. Online tools (TargetScan, miRTarBase, miRDB, and miRWalk) were used to predict the target gene of miR-101, and KOBAS was used for enrichment analysis of GO and KEGG pathways. The mimics and inhibitors of miR-101 were transfected into bovine testicular sertoli cells. Cell proliferation, apoptosis, secretion-related gene expression, and cell proliferation coefficient were detected by RT-qPCR, Western Blotting, and CCK-8 techniques.There were 26 common target genes for bta-miR-101 predicted by 4 online tools. GO and KEGG enrichment analysis showed that these genes were mainly enriched in the nucleoplasm, protein-binding, and JAK-STAT receptor signaling pathways, which were associated with cell differentiation, cell cycle, apoptosis, and other biological processes to varying degrees. The results of RT-qPCR showed that bta-miR-101 expression was significantly higher in testis tissue at birth than at sexual maturity (P<0.05), consistent with previous laboratory sequencing results. After transfection of miR-101 mimics and inhibitors, it was found that miR-101 mimics could promote the expression of proliferation-related genes at mRNA and protein levels, significantly increase the proliferation coefficient of sertoli cells, and inhibit the expression of apoptosis genes at mRNA and protein levels (P<0.05); miR-101 inhibitors inhibited the expression of proliferation-related genes at mRNA and protein levels and promoted the expression of apoptotic genes at mRNA and protein levels (P<0.05). In terms of cell secretion, miR-101 had an effect on the transcription of secretion marker genes GDNF and BMP4, but had no significant effect on the secretion of androgen-binding proteins. In conclusion, miR-101 promoted proliferation and inhibited apoptosis of immature sertoli cells of the bovine testis, and had no significant effect on the secretion of androgen-binding proteins.

Key words: bovine testicular sertoli cell, miR-101, proliferation, apoptosis

CLC Number:

S823.3

HU Qiaoyan, ZHAI Xiangqin, LI Yidan, HAN Jiale, LEI Chuzhao, DANG Ruihua. Effects of bta-miR-101 on Proliferation, Apoptosis and Secretion of Bovine Testicular Sertoli Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1040-1051.

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Effects of bta-miR-101 on Proliferation, Apoptosis and Secretion of Bovine Testicular Sertoli Cells

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