【摘要】：目的:選取自備的鋱摻雜的精氨酸表面修飾的羥基磷灰石(arginine functionalized hydroxyapatite,HAP/Arg)納米顆粒,研究該納米顆粒與細胞的相互作用及血管內皮細胞對HAP/Arg納米顆粒的內吞動力學和內吞入胞的相關機制。方法:選取人臍靜脈內皮細胞(human umbilical vein endothelial cells,HUVECs)作為研究模型細胞,利用激光掃描共聚焦顯微鏡觀察HAP/Arg納米顆粒的細胞攝取,利用細胞流式儀及原子力顯微鏡測定不同濃度下細胞攝取后的平均熒光強度。結果:鋱摻雜的HAP/Arg納米顆粒被HUVECs攝取后主要存在于細胞質中,而且大部分分布在細胞核區(qū)域周圍。HAP/Arg納米顆粒的入胞過程具有時間和濃度依賴性,作用時間以4 h為宜,納米顆粒的濃度以50 mg/L為最佳。精氨酸修飾的HAP納米顆粒入胞量明顯強于未經精氨酸修飾的HAP納米顆粒。結論:HAP/Arg納米顆粒進入HUVECs是一個主動轉運的能量依賴的過程,以網格蛋白和小凹蛋白介導的內吞相結合的細胞跨膜轉運機制,但以小凹蛋白介導內吞途徑為主。
[Abstract]:Objective: to select the terbium doped arginine surface modified hydroxyapatite (arginine functionalized hydroxyapatite,HAP/Arg) nanoparticles. The interaction between the nanoparticles and cells and the endocytosis kinetics of vascular endothelial cells to HAP/Arg nanoparticles and the mechanism of endocytosis were studied. Methods: human umbilical vein endothelial cells (human umbilical vein endothelial cells,HUVECs) were selected as model cells. The uptake of HAP/Arg nanoparticles was observed by laser scanning confocal microscopy. Cell flow cytometry and atomic force microscope (AFM) were used to measure the average fluorescence intensity of cells at different concentrations. Results: the terbium doped HAP/Arg nanoparticles mainly existed in the cytoplasm after HUVECs uptake, and most of them were located around the nuclear region. The entering process of HAP / Arg nanoparticles was in a time-and concentration-dependent manner, and the appropriate time of action was 4 h. The best concentration of nanoparticles is 50 mg/L. The number of arginine modified HAP nanoparticles was significantly higher than that of unarginine modified HAP nanoparticles. Conclusion the entry of HUVECs nanoparticles into HUVECs is an energy dependent process of active transport, which is mediated by griddle protein and concave protein mediated transmembrane transport mechanism, but mainly by concave protein mediated endocytosis pathway.
【作者單位】： 中南大學湘雅三醫(yī)院醫(yī)學實驗中心;中南大學醫(yī)用材料與器械研究中心;
【基金】：湖南省科技廳重點研發(fā)計劃項目(2016JC2064) 教育部博士學科點基金(20130162120094) 中南大學粉末冶金國家重點實驗室開放課題~~
【分類號】：R318.08

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