摘要
本试验旨在探明小檗碱(Berberine,Ber)对猪卵母细胞体外成熟的作用及其对miRNA表达差异和脂滴代谢相关基因的影响。吸取法获取卵丘-卵母细胞复合体,0.1%透明质酸酶脱除颗粒细胞的未成熟卵母细胞为GV组,以TCM-199培养液为体外成熟液作为对照组,添加中药单体成分Ber为Ber组,观察卵母细胞成熟效果,基因芯片技术分析Ber对猪卵母细胞体外成熟过程中miRNA的差异变化,尼罗红染色测定脂滴含量变化,RT-PCR检测候选基因及脂质代谢相关基因表达含量。结果表明:Ber组卵母细胞颗粒细胞扩散率(P<0.05)和第一极体排出率较对照组均差异显著(P<0.01);相比于对照组和GV组间的差异miRNA,Ber组上调4个在卵母细胞成熟中差异表达的miRNA,下调1个miRNA;Ber组卵母细胞脂滴含量明显少于对照组;Ber组卵母细胞中与脂质代谢相关的基因的表达均极显著低于对照组(P<0.01),且对照组卵母细胞极显著低于GV组(P<0.01)。Ber可能通过影响miRNA及脂质代谢相关的基因,降低猪卵母细胞中脂滴的含量,进而促进猪卵母细胞体外成熟。
To study the effects of berberine （Ber） on miRNA and related genes of lipid metabolism in porcine oocytes matured in vitro. Obtaining the cumulus oocyte complexes by the draw method, 0.1% hyaluronidase to remove granulosa cell of the immature oocytes as the GV group, GV porcine oocytes undergo in vitro maturation 44h, TCM-199 as control group and adding berberine as Ber group respectively, these groups oocytes were observed effects of matured in vitro and collected oocytes of GV, control and Ber group to proceed gene chip analysis. The content of lipid droplets were measured by Nile red staining, and RT-PCR was used to measure the related gene expression of lipid metabolism. Comparing with the control group, we found that granular cell proliferation rate （P〈0.05） and the polar body I （PbI）formation rate （P〈0.01） were significantly increased. Comparing with the different expression miRNA of control group vs GV, Ber group up-regulated 4 miRNA and down-regulated 1 miRNA, and the GO and pathway analysis found that these miRNA mainly involved in metabolic process; The oocytes lipid content of Ber group significantly lower than that of the control; The lipid metabolism related genes expression all significantly lower than the control group （P〈0.01） and the control group significantly lower than GV oocytes （P〈0.01）. Berberine through regulating miRNA and the related genes of lipid metabolism deceased porcine oocytes lipid content and improved the porcine oocytes matured in vitro.
出处
《中国畜牧杂志》
CAS
北大核心
2017年第10期47-51,95,共6页
Chinese Journal of Animal Science
基金
国家自然基金项目(31572402)
科技创新服务能力建设-科研计划(市级)重点项目(KZ201610020017)
