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小檗碱通过 miR-192 调节体外成熟猪卵母细胞的脂质代谢。

Berberine regulates lipid metabolism via miR-192 in porcine oocytes matured in vitro.

机构信息

Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.

College of Animal Science and Technology, China Agricultural University, Beijing, China.

出版信息

Vet Med Sci. 2021 May;7(3):950-959. doi: 10.1002/vms3.393. Epub 2021 Apr 5.

Abstract

BACKGROUND

The berberine (Ber) is an isoquinoline alkaloid compound extracted from Rhizoma coptidis and has the effect that reduces adipose. MicroRNA-192 (miR-192) is related to fat metabolism. However, the relevant mechanism of berberine on lipid metabolism during in vitro maturation (IVM) of porcine oocytes remains unclear.

OBJECTIVES

In this study, we investigated the molecular mechanism by which berberine promotes the IVM and lipid metabolism of porcine oocytes via miR-192.

METHODS

Ber was added to IVM medium of porcine oocytes. MiR-192 agomir, miR-192 antagomir and negative control fragment were microinjected into the cytoplasm of oocytes without Ber. Rates of oocyte IVM and embryonic development in each group were observed. The content of lipid droplets in IVM oocytes in each group was analyzed by Nile red staining. Expression levels of miR-192 and FABP3, SREBF1 and PPARG, were detected by qPCR and western blotting. The target genes of miR-192 were determined by luciferase reporter assays.

RESULTS AND CONCLUSIONS

We found that Ber significantly increased the rate of oocytes IVM and blastocyst development, and decreased the area and numbers of lipid droplets in IVM oocytes. Ber significantly increased the expression of miR-192 in IVM oocytes, and significantly decreased the expression of SREBF1 and PPARG, which were target genes of miR-192. This study indicates that Ber promotes lipid metabolism in porcine oocytes by activating the expression of miR-192 and down-regulating SREBF1 and PPARG, thus, improving IVM of porcine oocytes.

摘要

背景

小檗碱(Ber)是从黄连中提取的一种异喹啉生物碱化合物,具有减少脂肪的作用。MicroRNA-192(miR-192)与脂肪代谢有关。然而,小檗碱在猪卵母细胞体外成熟(IVM)过程中对脂质代谢的相关机制尚不清楚。

目的

本研究旨在探讨小檗碱通过 miR-192 促进猪卵母细胞 IVM 及脂质代谢的分子机制。

方法

在猪卵母细胞 IVM 培养基中添加小檗碱。将 miR-192 agomir、miR-192 antagomir 和阴性对照片段微注射到没有小檗碱的卵母细胞胞质中。观察各组卵母细胞 IVM 和胚胎发育的比例。通过尼罗红染色分析各组 IVM 卵母细胞中脂滴的含量。通过 qPCR 和 Western blot 检测 miR-192 及 FABP3、SREBF1 和 PPARG 的表达水平。通过荧光素酶报告基因实验确定 miR-192 的靶基因。

结果与结论

结果发现,小檗碱显著提高了卵母细胞 IVM 和囊胚发育的比例,降低了 IVM 卵母细胞中脂滴的面积和数量。小檗碱显著增加了 IVM 卵母细胞中 miR-192 的表达,并显著降低了 miR-192 的靶基因 SREBF1 和 PPARG 的表达。该研究表明,小檗碱通过激活 miR-192 的表达并下调 SREBF1 和 PPARG 的表达,促进猪卵母细胞的脂质代谢,从而改善猪卵母细胞的 IVM。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6c6/8136937/3e2950fbfcd7/VMS3-7-950-g004.jpg

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