Khongwichit Soemwit, Swangphon Piyawut, Nualla-Ong Aekkaraj, Prompat Napat, Amatatongchai Maliwan, Lieberzeit Peter A, Chunta Suticha
Faculty of Medical Technology, Prince of Songkla University, Songkhla 90110, Thailand.
Division of Biological Science, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand.
ACS Appl Bio Mater. 2025 Jan 20;8(1):457-474. doi: 10.1021/acsabm.4c01432. Epub 2025 Jan 6.
The accumulation of oxidized low-density lipoprotein (oxLDL) in macrophages leads to the formation of foam cells and atherosclerosis development. Reducing the uptake of oxLDL in macrophages decreases the incidence and progression of atherosclerosis. Four distinct single-strand DNA sequences, namely, AP07, AP11, AP25, and AP29, were selected that demonstrated specific binding to distinct regions of oxidized apolipoprotein B100 (apoB100; the protein component of oxLDL) with low HDOCK scores. These four DNA sequences were combined to generate aptamers that selectively bound to labeled Dil-oxLDL, and were subsequently added to murine RAW 264.7 macrophages to test their inhibitory effects using fluorescence spectrometry. The four combined aptamers at 10 μM reduced oxLDL uptake by 79 ± 4% compared to that of the untreated aptamer group. Flow cytometry data demonstrated that macrophages treated with aptamers reached only 32.6% of the Dil-oxLDL signal, a 50% reduction in fluorescence emission relative to that of the untreated group (64.4% Dil-oxLDL signal). Binding the four combined aptamers to the oxLDL surface disrupted the interaction between oxLDL and CD36 via cyclic voltammetry, effectively decreasing the level of uptake of oxLDL by macrophages. Results suggested that these aptamers could be used as alternative compounds to prevent the formation of foam cells, hence providing antiatherosclerosis activity.
巨噬细胞中氧化型低密度脂蛋白(oxLDL)的积累会导致泡沫细胞的形成和动脉粥样硬化的发展。减少巨噬细胞对oxLDL的摄取可降低动脉粥样硬化的发生率和进展。选择了四个不同的单链DNA序列,即AP07、AP11、AP25和AP29,它们以低HDOCK分数显示出与氧化载脂蛋白B100(apoB100;oxLDL的蛋白质成分)的不同区域特异性结合。将这四个DNA序列组合以生成与标记的Dil-oxLDL选择性结合的适体,随后将其添加到小鼠RAW 264.7巨噬细胞中,使用荧光光谱法测试它们的抑制作用。与未处理的适体组相比,10 μM的四种组合适体使oxLDL摄取减少了79±4%。流式细胞术数据表明,用适体处理的巨噬细胞仅达到Dil-oxLDL信号的32.6%,相对于未处理组(64.4% Dil-oxLDL信号)荧光发射降低了50%。通过循环伏安法将四种组合适体与oxLDL表面结合破坏了oxLDL与CD36之间的相互作用,有效降低了巨噬细胞对oxLDL的摄取水平。结果表明,这些适体可作为预防泡沫细胞形成的替代化合物,从而提供抗动脉粥样硬化活性。