可逆抑制血管内皮生长因子(VEGF)表达转基因小鼠子宫基因转录谱分析

Transcriptional profiling of mouse uterus at pre-implantation stage under VEGF repression

目的:研究血管内皮生长因子(VEGF)对子宫中基因表达水平及信号通路的调控作用。方法:利用四环素可逆调节VEGF基因表达的转基因小鼠,通过新一代测序数字基因表达谱(DGE)方法分析在VEGF表达被抑制情况下的子宫基因表达水平。采用实时定量PCR验证部分表达异常的基因。结果:检测到在小鼠子宫中表达的基因约6×106个。其中在VEGF基因表达被抑制条件下,表达水平变化比较明显的基因有2 398个,其中上调的基因为1 231个,主要包括调节细胞内激酶基因、细胞分裂的基因、蛋白折叠的基因等;下调的基因为1 167个,主要包括调节肌肉发育的基因、细胞内基质形成的基因、血管形成的基因等。荧光定量PCR验证表明,基因表达水平变化与转录谱结果高度相关,VEGF抑制后小鼠子宫内表达上调的基因有P2ry14、Eln、Rnf187、Oxtr、Inhbb、Pabpn1、Car2和Mif1(r=0.972),表达下调的基因VEGF、Acta2、Pcx、Tnc、Igfbp3、Cnn1和Hopx(r=0.931)。结论:VEGF表达水平与子宫功能的维持密切相关,为不明原因不孕的诊治提供了新思路。

Objective： To study the value of vascular endothelial growth factor （VEGF） in reproduction immunology. Methods： The transgenic mouse model of tetracycline controlled reversible VEGF repression was established. Digital gene expression （DGE） method was used to to analyze gene expression level changes of mouse uterus. Partly genes with abnormal expression was identified by Real-time PCR. Results： There were about 6 million genes expressed in mouse uterus. Under the condition of VEGF repression, 2 398 genes expression were changed over 200 folds compared with the control. A total of 1 231 genes were up-regulated, include genes regulating intracellular kinase, cell devision, protein folding, etc. While 1 167 genes were down-regulated, including genes regulating muscle development, cell matrix construction, blood vessel forming, etc. Expression of up- or down- regulated gene by VEGF repression in DGE analysis were highly associated, indicated by Real-time PCR. After VEGF repression, up-regulated genes included P2ry14, Eln, Rnf187, Oxtr, Inhbb, Pabpn 1, Car2 and Mill （r=0.972）, down-regulated genes included VEGF, Acta2, Pcx, Tnc, Igfbp3, Cnn l and Hopx （r=0.931）. Conclusion： VEGF expression level is associated with reproductive function, which may become a new target of treating infertility diseases.