miR-148a-3p靶向PPARγ基因抑制鹅颗粒细胞孕酮的合成

doi:10.11843/j.issn.0366-6964.2021.06.012

Abstract

Abstract: The aim of this study was to explore whether miR-148a-3p could target PPARγ to regulate the production of steroid hormone in granulosa cells of goose. In the present study, 12 healthy and high-laying maternal Tianfu meat female geese were used for sample collection and culture of granulosa cells in vitro. Quantitative polymerase chain reaction (qPCR) was used to detect the expression profile of miR-148a-3p at different stages of granulosa layers in follicles of goose. MEGA 7 software was used to identify the conservation of miR-148a-3p in different species. Target genes of miR-148a-3p were predicted and validated via double luciferase report system. miR-148a-3p mimics and inhibitor were transfected into goose granulosa cells, and the expression levels of target gene and downstream genes were examined by qPCR. Meanwhile, ELISA was used to measure the changes of hormone levels. The results showed that miR-148a-3p was highly conserved in different species. The expression level of miR-148a-3p firstly increased and then decreased during the follicles development of goose. In the culture of goose granulosa cells in vitro, the expression level of 3β-HSD and the progesterone production significantly decreased after the transfection of miR-148a-3p mimics(P<0.05), while the above indicators increased after the transfection of miR-148a-3p inhibitor (P<0.05). Bioinformatic analysis and double luciferase report system demonstrated that PPARγ was the target gene of miR-148a-3p. The expression level of PPARγ significantly decreased after transfection of miR-148a-3p mimics in granulosa cells(P<0.05), while that increased after transfection of miR-148a-3p inhibitor(P<0.05). After interfering PPARγ, the expression level of 3β-HSD significantly decreased(P<0.05), and the progesterone production also decreased. These results demonstrated that miR-148a-3p could target PPARγ to inhibit the expression of 3β-HSD and decrease the progesterone production in granulosa cells of goose.

Key words: miR-148a-3p, PPARγ, goose, granulosa cells, progesterone, 3β-HSD

CLC Number:

S835.2

DENG Yan, XIE Guangjuan, HU Shenqiang, LI Liang, WANG Jiwen. miR-148a-3p Inhibits the Production of Progesterone by Targeting PPARγ in Granulosa Cells of Goose[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1571-1581.

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miR-148a-3p Inhibits the Production of Progesterone by Targeting PPARγ in Granulosa Cells of Goose

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