Jaakkola O, Ylä-Herttuala S, Särkioja T, Nikkari T
Department of Biomedical Sciences, University of Tampere, Finland.
Atherosclerosis. 1989 Oct;79(2-3):173-82. doi: 10.1016/0021-9150(89)90122-6.
Intimal cells from human aortic fatty streak lesions were isolated with collagenase-elastase digestion and the cellular uptake of lipoproteins fluorescently labeled with 3,3'-dioctadecylindocarbocyanine (DiI) was studied in primary culture. The majority of the cells in primary culture contained lipid droplets and the foam cells consisted of both macrophages and smooth muscle cells (SMC), identified with electron microscopy and the macrophages also using the monoclonal anti-Leu-M3 antibody. The lipid inclusions contained cholesteryl ester, as visualized with filipin staining. Arterial macrophages took up DiI-labeled acetylated low density lipoprotein (DiI-acetyl-LDL) in the same way as did monocyte-macrophages isolated from blood. DiI-labeled beta-very low density lipoprotein (DiI-beta-VLDL) isolated from cholesterol-fed rabbits, was taken up by both macrophages and SMCs. In macrophages DiI-beta-VLDL was internalized also in the presence of excess unlabeled low density lipoprotein (LDL), whereas in SMCs the uptake was partially prevented. DiI-LDL uptake was only seen in SMCs free of lipid inclusions and especially during cell growth. The present results show that, in human aortic fatty streaks, (a) both macrophages and SMCs accumulate cholesteryl ester, (b) macrophage foam cells possess active scavenger receptors capable of mediating the uptake of acetyl-LDL, and (c) macrophages are also capable of accumulating cholesteryl ester by receptor-mediated uptake of beta-VLDL.
采用胶原酶-弹性蛋白酶消化法分离人主动脉脂肪条纹病变的内膜细胞,并在原代培养中研究用3,3'-二辛基吲哚碳菁(DiI)荧光标记的脂蛋白的细胞摄取情况。原代培养中的大多数细胞含有脂滴,泡沫细胞由巨噬细胞和平滑肌细胞(SMC)组成,通过电子显微镜鉴定,巨噬细胞也使用单克隆抗-Leu-M3抗体进行鉴定。用制霉菌素染色可见脂质内含物含有胆固醇酯。动脉巨噬细胞摄取DiI标记的乙酰化低密度脂蛋白(DiI-乙酰-LDL)的方式与从血液中分离的单核细胞-巨噬细胞相同。从喂食胆固醇的兔子中分离出的DiI标记的β-极低密度脂蛋白(DiI-β-VLDL)被巨噬细胞和SMC摄取。在巨噬细胞中,DiI-β-VLDL在存在过量未标记的低密度脂蛋白(LDL)的情况下也被内化,而在SMC中摄取则部分受到抑制。DiI-LDL摄取仅在没有脂质内含物的SMC中观察到,尤其是在细胞生长期间。目前的结果表明,在人主动脉脂肪条纹中,(a)巨噬细胞和SMC都积累胆固醇酯,(b)巨噬细胞泡沫细胞具有能够介导乙酰-LDL摄取的活性清道夫受体,(c)巨噬细胞也能够通过受体介导的β-VLDL摄取积累胆固醇酯。