【摘要】：目的用新型聚乙烯亞胺納米凝膠(PEI-NP)攜帶小RNA(miRNA)質粒沉默大鼠Kupffer細胞(KC)核因子κB(NF-κB)的表達。方法凝膠電泳檢測PEI-NP裝載miRNA質粒的容量;PEI-NP/miRNA質粒轉染RAW264.7細胞后分別采用CCK-8法檢測細胞毒性,annexinⅤ-FITC/PI雙染色結合流式細胞術檢測RAW264.7細胞凋亡和轉染率,實時定量PCR(qRT-PCR)檢測RAW264.7細胞NF-κB mRNA水平,Western blot法檢測RAW264.7細胞NF-κB蛋白水平,透射電鏡觀察陽離子聚合物在RAW264.7細胞和肝組織KC內的分布;免疫組織化學技術檢測KC內NF-κB的表達。結果 PEI-NP/miRNA質粒質量比為20∶1時載荷率接近100%,體外轉染RAW264.7細胞48 h,轉染率為(33.63±1.94)%。與對照組相比,PEI-NP/miRNA轉染的RAW264.7細胞NF-κB蛋白含量顯著降低,透射電鏡觀察到RAW264.7細胞內有大量PEI-NP。肝臟組織透射電鏡只在KC內觀察到PEI-NP分布;與對照組相比,免疫組織化學染色結果顯示,KC內NF-κB蛋白水平明顯降低。結論 PEI-NP攜帶miRNA質粒能成功轉染體外培養(yǎng)的巨噬細胞和體內肝臟KC并沉默NF-κB的表達。
[Abstract]:Aim to silence the expression of (KC) nuclear factor 魏 B (NF- 魏 B) in rat Kupffer cells using a novel polyvinyleneimine nanogel (PEI-NP) carrying small RNA (miRNA) plasmid. Methods the capacity of PEI-NP loaded with miRNA plasmid was detected by gel electrophoresis. After transfection of PEI-NP/miRNA plasmid into RAW264.7 cells, cytotoxicity was detected by CCK-8 assay, apoptosis and transfection rate of RAW264.7 cells were detected by annexin 鈪，

本文編號：2381170