School of Food and Biological Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, People's Republic of China.
J R Soc Interface. 2012 Jun 7;9(71):1233-40. doi: 10.1098/rsif.2011.0436. Epub 2011 Nov 2.
To clarify the effect of concentration polarization of oxidative modification of low-density lipoproteins (ox-LDLs) on human smooth muscle cells (SMCs), the proliferation, ox-LDL uptake and apoptosis with SMCs cultured on permeable (the permeable group) or non-permeable membranes (the non-permeable group) were analysed by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay, spectrofluorometry and flow cytometry using a parallel-plate flow chamber technique. The concentration polarization of ox-LDLs at the surface of the cultured cell monolayer was assessed by confocal laser microscopy. The results showed that concentration polarization of ox-LDLs could indeed occur at the cultured cell monolayer surface of the permeable group, leading to an enhanced wall concentration of ox-LDLs that was over 15 per cent higher than the bulk concentration of the perfusion solution at a pressure of 100 mmHg. When concentration of ox-LDLs in the perfusion solution was less than or equal to 100 µg ml(-1), SMCs' proliferation was induced, while cell apoptosis was induced when its concentration was above 150 µg ml(-1). The uptake of ox-LDLs by the cultured cells was significantly higher for the permeable group than for the non-permeable group. In addition, the ox-LDL-induced cell death and apoptosis were much more severe in the permeable group than that in the non-permeable group. Therefore, the experimental study suggests that concentration polarization of ox-LDLs plays an adverse role in the vascular system owing to its toxicity to vascular cells, in turn enhance ox-LDL infiltration into the arterial wall and accelerate SMC apoptosis.
为了阐明低密度脂蛋白(LDL)氧化修饰的浓差极化对人平滑肌细胞(SMC)的影响,我们通过 3-(4,5-二甲基噻唑基-2)-2,5-二苯基四氮唑溴盐(MTT)比色法、荧光分光光度法和流式细胞术,分析了在可渗透(渗透组)或不可渗透(不可渗透组)膜上培养的 SMC 的增殖、ox-LDL 摄取和凋亡情况,采用平行板流动室技术。通过共聚焦激光显微镜评估 ox-LDL 在培养细胞单层表面的浓差极化。结果表明,ox-LDL 确实可以在渗透组培养的细胞单层表面发生浓差极化,导致 ox-LDL 的壁浓度比灌注溶液的体浓度高 15%以上,压力为 100mmHg。当灌注液中 ox-LDL 的浓度小于或等于 100μg/ml 时,SMC 增殖被诱导,而当其浓度高于 150μg/ml 时,细胞凋亡被诱导。培养细胞对 ox-LDL 的摄取在渗透组明显高于不可渗透组。此外,ox-LDL 诱导的细胞死亡和凋亡在渗透组比不可渗透组更为严重。因此,实验研究表明,由于 ox-LDL 对血管细胞的毒性,其在血管系统中的浓差极化会产生不良作用,从而增强 ox-LDL 渗透到动脉壁并加速 SMC 凋亡。