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小檗碱对猪体外受精胚胎发育的作用及微小RNA的差异表达分析

Function of berberine on porcine in vitro fertilization embryo development and differential expression analysis of microRNAs.

作者信息

Wang Jun-Li, Zhang Chao, Liu Bing, Huang Xiao-Meng, Dai Jia-Ge, Tian Jian-Hui, Gao Jian-Ming

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

Reprod Domest Anim. 2019 Mar;54(3):520-530. doi: 10.1111/rda.13397. Epub 2019 Feb 18.

Abstract

The effect of berberine (Ber) on in vitro fertilization (IVF) embryo development in pigs and the associated differential expression of microRNAs (miRNAs) in the embryo were investigated. NCSU-23 embryonic culture medium was used for a control group, while NCSU-23 embryonic culture medium added with Ber was used for a Ber group. The embryo development rates in these groups were determined, and the zygotes, 4- and 8-cell embryos, and blastocysts were collected for cDNA microarray analysis. The development rates of 2-, 4-, 8-cell embryos and blastocysts were significantly higher in the Ber group than those in the control group (p < 0.01). The differentially expressed miRNAs in the 8-cell versus the 4-cell stage in control group as well as in the 8-cell Ber group versus the 8-cell control group overlapped, and it was found that nine miRNAs were commonly upregulated and two of them were downregulated, while there was no overlap among the other groups. The target genes of Ber-regulated miRNAs at the 8-cell stage were mainly associated with the molecular pathway of nucleic acid and protein synthesis. These findings suggest that Ber may regulate the expression of miRNAs at the 8-cell stage, which is beneficial to provide material reserves for the maternal to zygote transition of porcine embryos, thereby increasing the porcine IVF embryo development rate.

摘要

研究了黄连素(Ber)对猪体外受精(IVF)胚胎发育的影响以及胚胎中微小RNA(miRNA)的相关差异表达。对照组使用NCSU-23胚胎培养基,而添加Ber的NCSU-23胚胎培养基用于Ber组。测定这些组的胚胎发育率,并收集合子、4细胞和8细胞胚胎以及囊胚用于cDNA微阵列分析。Ber组中2细胞、4细胞、8细胞胚胎和囊胚的发育率显著高于对照组(p < 0.01)。对照组中8细胞与4细胞阶段以及Ber组8细胞与对照组8细胞之间差异表达的miRNA存在重叠,发现9个miRNA共同上调,其中2个下调,而其他组之间无重叠。Ber调控的8细胞阶段miRNA的靶基因主要与核酸和蛋白质合成的分子途径相关。这些发现表明,Ber可能在8细胞阶段调节miRNA的表达,这有利于为猪胚胎从母体向合子的转变提供物质储备,从而提高猪IVF胚胎发育率。

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