發(fā)布時間：2018-05-21 15:49

  本文選題：血管內(nèi)皮生長因子(VEGF) + 血管內(nèi)皮生長因子受體��； 參考：《南方農(nóng)業(yè)學(xué)報》2017年02期

【摘要】：【目的】明確RNA干擾(RNAi)對綿羊顆粒細(xì)胞體外培養(yǎng)過程中血管內(nèi)皮生長因子(Vascular endothelial growth factor,VEGF)及其受體mRNA表達的影響,為開展VEGF在綿羊卵母細(xì)胞體外成熟和胚胎發(fā)育過程中信號通路的相關(guān)研究打下基礎(chǔ)�！痉椒ā坎捎秒姶┛准夹g(shù)將針對VEGF兩個受體Flt-1和KDR/Flk-1的兩條有效雙鏈小RNA導(dǎo)入綿羊顆粒細(xì)胞內(nèi),利用實時熒光定量PCR檢測體外培養(yǎng)綿羊顆粒細(xì)胞各培養(yǎng)時間VEGF、Flt-1和KDR/Flk-1 mRNA表達水平的變化。設(shè)定篩選出的Flt-1和KDR/Flk-1有效干擾片段分別為試驗組Ⅰ和試驗組Ⅱ,兩個干擾片段共同作用為試驗組Ⅲ,無效的干擾片段為對照組�！窘Y(jié)果】綿羊顆粒細(xì)胞體外培養(yǎng)過程中,VEGF mRNA在各時間段的相對表達量是Flt-1 mRNA相對表達量的102倍、KDR/Flk-1 mRNA相對表達量的103倍;培養(yǎng)第1 d時,試驗組Ⅰ、試驗組Ⅱ和試驗組ⅢVEGF mRNA相對表達量分別為2.62×10-2、3.54×10-2和2.94×10-2,均極顯著高于對照組(P0.01,下同),3個試驗組的VEGF mRNA表達量從第2 d開始降低,直到第6 d與對照組趨于一致。試驗組ⅡFlt-1 mRNA相對表達量從培養(yǎng)第1 d的1.02×10-1逐漸降至第7 d的8.99×10-4,且一直極顯著高于其他組;試驗組Ⅰ和試驗組Ⅲ在前3 d幾乎無Flt-1 mRNA表達,隨著培養(yǎng)時間的延長逐漸呈上升趨勢,第8 d時各組趨于一致。試驗組Ⅰ的KDR/Flk-1 mRNA相對表達量從第1 d開始極顯著低于對照組;試驗組Ⅱ和試驗組Ⅲ在前3 d幾乎無KDR/Flk-1 mRNA相對表達,隨著培養(yǎng)時間的延長呈上升趨勢,第9 d時各組趨于一致。【結(jié)論】兩個有效干擾片段在綿羊顆粒細(xì)胞體外培養(yǎng)過程中起到很好的干擾作用,可改變VEGF、Flt-1及KDR mRNA的表達量,進而影響顆粒細(xì)胞相關(guān)生長信號的傳輸。
[Abstract]:[objective] to investigate the effect of RNA interference RNAi on the expression of vascular endothelial growth factor (VEGF) and its receptor (mRNA) in sheep granulosa cells in vitro. To lay a foundation for the study of VEGF signaling pathway in ovine oocytes during in vitro maturation and embryonic development. [methods] two effective double-stranded small RNA targeting VEGF receptor Flt-1 and KDR/Flk-1 were introduced by electroporation technique. In sheep granulosa cells, The expression of VEGF Flt-1 and KDR/Flk-1 mRNA in sheep granulosa cells were detected by real-time fluorescence quantitative PCR. The effective interference fragments of Flt-1 and KDR/Flk-1 were selected as experimental group 鈪，

本文編號：1919824