[ Objective] This study was to investigate the effect of VEGF and its receptor Fit-1 mRNA expression in Mongolia sheep umbilical vein endothelial cells by ghrelin antisense inhibition. [ Method] Experiments were divided into 4 groups： group Ⅰ （blank control group） ; group Ⅱ （liposome group） ; group Ⅲ （SCON group： 20 μmol/L sense oligonucleotide） ; group Ⅳ （ASCON： 20 μmol/L antisense oligonucleotide）. VEGF and its receptor Fit-1 mRNA expression changes were detected by using real-time fluorescence quantitative detection after 24, 36 and 48 h. [ Result] The expression of VEGF mRNA in group Ⅰ, group Ⅱ were insignificantly different at higher expression levels, and did not change significantly with the time; the expression of VEGF mRNA in group Ⅲ assumed a slight decrease, but there were no significant differences between group I and group Ⅱ （P 〉0.05）, the expression of VEGF mRNA in group Ⅳ（antisense oligonucleotide group ） decreased significantly （P 〈 0.05） ; the expression of VEGF receptor FLT-1 mRNA was similar to that of VEGF. [ Conclusion] Antisense inhibition ghrelin has a downward effect to the expression of VEGF and its receptor Fit-1 the mRNA.
该研究旨在探讨外源Ghrelin对绵羊卵母细胞体外成熟的调控作用。在成熟培养液中分别添加浓度为0、100、200和300ng/m L的Ghrelin,进行绵羊卵母细胞体外培养,成熟培养24h,观察统计处于MII期的细胞数目。选择最佳添加浓度,在体外成熟培养0、6、8、14、16、24h,通过Hoechst33342染色持续检测卵细胞中细胞核成熟随时间变化,并对不同培养时期细胞进行实时荧光定量PCR检测,以证明细胞质量相关基因的相对表达量。成熟培养24h时,添加0、100、200、300 ng/m LGhrelin组卵母细胞成熟率分别为:63.7%(220/345)、69.4%(230/332)、85.6%(270/316)、75.9%(231/305)。200ng/m L Ghrelin组显著高于对照组(P<0.01),其余组并未显著高于对照组(P<0.05);染色证明:200ng/m L Ghrelin组于成熟培养0h、6h、14h、16h到达GV、GVBD、MI、MII期(P<0.05);实时荧光定量PCR证明:对卵母细胞成熟GV、GVBD、MI、MII期,Ghrelin受体基因GH-SR-1a和卵母细胞质量相关基因OCT-4、GLUT1和IFNT的相对表达时间提前,但并不显著影响这些基因的表达水平。外源Ghrelin参与了绵羊卵母细胞的体外成熟调控,可在没有改变其成熟过程的前提下加速卵母细胞的体外成熟过程,但对绵羊卵母细胞的质量和进一步发育相关的基因的表达没有明显影响。
