摘要
[ 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.
[目的]探讨ghrelin对蒙古绵羊脐静脉内皮细胞的血管内皮生长因子VEGF及其受体Flt-1mRNA表达的影响。[方法]试验分为4组:I组,空白对照组;II组,脂质体组(不加寡核苷酸);III组(SCON),20μmol/L正义寡核苷酸组;IV组(ASCON),20μmol/L反义寡核苷酸组。上述各组均在培养24、36、48h后,采用实时荧光定量方法检测VEGF及其受体Flt-1mRNA表达情况的变化。[结果]VEGF和Flt-1熔解曲线表明扩增效果好,培养24h,VEGFmRNAI组相对表达量为0.0769,II组为0.0768,III组为0.0769,3组之间无显著性差异(P>0.05),IV组VEGFmRNA相对表达量下降为0.0242,且与I、II、III组有显著性差异(P<0.05)。培养36h,前3组相对表达量分别为0.059,0.051,0.0514,3组之间无显著性差异(P>0.05),第IV组为0.0242,与前3组比较有显著性差异(P<0.05)。培养48h,前3组相对表达量分别为0.061,0.0552,0.0521,3组之间无显著性差异(P<0.05),第IV组为0.0115,与前3组比较有显著性差异(P<0.05)。培养24h,Flt-1mRNAI组相对表达量为0.0786;II组为0.0782,III组为0.0781,3组之间无显著性差异(P>0.05),IV组Flt-1mR-NA相对表达量下降为0.0256,且与I、II、III组有显著性差异(P<0.05)。培养36h,前3组相对表达量分别为0.0781,0.0784,0.0782,3组之间无显著性差异(P>0.05),第IV组为0.0228,与前3组比较有显著性差异(P<0.05)。培养48h,前3组相对表达量分别为0.0691,0.0780,0.0693,3组之间无显著性差异(P>0.05),第IV组为0.0215,与前3组比较有显著性差异(P<0.05)。可见,VEGF受体Flt-1mRNA的表达与VEGF相似。二者均在I组、II组表达量无差异,具有较高的表达量,随着时间的延长没有明显变化;在III组表达轻微下降,但与I组和II组无显著性差异(P>0.05);IV组的表达量显著下降(P<0.05),并且随着时间的延长至逐渐下降。[结论]反义抑制ghrelin对VEGF及其受体Flt-1的mRNA表达有下调作用。
基金
Supported by National Natural Science Foundation of China(30860201)~~
