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

【摘要】：【目的】明確RNA干擾(RNAi)對(duì)綿羊顆粒細(xì)胞體外培養(yǎng)過(guò)程中血管內(nèi)皮生長(zhǎng)因子(Vascular endothelial growth factor,VEGF)及其受體mRNA表達(dá)的影響,為開(kāi)展VEGF在綿羊卵母細(xì)胞體外成熟和胚胎發(fā)育過(guò)程中信號(hào)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(duì)VEGF兩個(gè)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(shí)時(shí)熒光定量PCR檢測(cè)體外培養(yǎng)綿羊顆粒細(xì)胞各培養(yǎng)時(shí)間VEGF、Flt-1和KDR/Flk-1 mRNA表達(dá)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(yàn)組Ⅰ和試驗(yàn)組Ⅱ,兩個(gè)干擾片段共同作用為試驗(yàn)組Ⅲ,無(wú)效的干擾片段為對(duì)照組�！窘Y(jié)果】綿羊顆粒細(xì)胞體外培養(yǎng)過(guò)程中,VEGF mRNA在各時(shí)間段的相對(duì)表達(dá)量是Flt-1 mRNA相對(duì)表達(dá)量的102倍、KDR/Flk-1 mRNA相對(duì)表達(dá)量的103倍;培養(yǎng)第1 d時(shí),試驗(yàn)組Ⅰ、試驗(yàn)組Ⅱ和試驗(yàn)組ⅢVEGF mRNA相對(duì)表達(dá)量分別為2.62×10-2、3.54×10-2和2.94×10-2,均極顯著高于對(duì)照組(P0.01,下同),3個(gè)試驗(yàn)組的VEGF mRNA表達(dá)量從第2 d開(kāi)始降低,直到第6 d與對(duì)照組趨于一致。試驗(yàn)組ⅡFlt-1 mRNA相對(duì)表達(dá)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(yàn)組Ⅰ和試驗(yàn)組Ⅲ在前3 d幾乎無(wú)Flt-1 mRNA表達(dá),隨著培養(yǎng)時(shí)間的延長(zhǎng)逐漸呈上升趨勢(shì),第8 d時(shí)各組趨于一致。試驗(yàn)組Ⅰ的KDR/Flk-1 mRNA相對(duì)表達(dá)量從第1 d開(kāi)始極顯著低于對(duì)照組;試驗(yàn)組Ⅱ和試驗(yàn)組Ⅲ在前3 d幾乎無(wú)KDR/Flk-1 mRNA相對(duì)表達(dá),隨著培養(yǎng)時(shí)間的延長(zhǎng)呈上升趨勢(shì),第9 d時(shí)各組趨于一致�！窘Y(jié)論】?jī)蓚�(gè)有效干擾片段在綿羊顆粒細(xì)胞體外培養(yǎng)過(guò)程中起到很好的干擾作用,可改變VEGF、Flt-1及KDR mRNA的表達(dá)量,進(jìn)而影響顆粒細(xì)胞相關(guān)生長(zhǎng)信號(hào)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(hào)：1919824