Hayavi Sima, Baillie George, Owens Moira D, Halbert Gavin W
Department of Pharmaceutical Sciences, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, UK.
J Pharm Pharmacol. 2006 Oct;58(10):1337-42. doi: 10.1211/jpp.58.10.0006.
Low density lipoprotein (LDL) is a normal plasma component, which is of interest in a number of research areas such as hypercholesterolaemia, drug targeting in cancer chemotherapy and as a lipid supplement in tissue culture systems. Currently, however, it can only be obtained by extraction from fresh plasma samples, which yields only small quantities. Synthetic LDL (sLDL) has been prepared using readily available lipid components coupled with a synthetic amphiphatic peptide molecule containing the apoprotein B receptor sequence. sLDL was capable of supporting the growth of Chinese Hamster Ovary (CHO) and fibroblast cells in serum-free culture media in a cholesterol-dependent manner that was related to the presence of the receptor peptide molecule. sLDL could be fluorescently labelled with 3,3'-dioctadecyloxalocarbocyanine perchlorate (DiO), and once labelled was assimilated by CHO and fibroblast cells in a time- and temperature-dependent manner that was dependent upon the presence of the receptor peptide. In addition, assimilation was reduced by an excess of unlabelled native LDL. The results indicated that the interaction of sLDL with CHO and fibroblast cells occurred via a receptor dependent system, most likely the LDL cellular receptor. sLDL is therefore a useful, easily obtained substitute for native LDL with potential applications in the areas of drug targeting to cells and serum-free tissue culture systems.
低密度脂蛋白(LDL)是一种正常的血浆成分,在多个研究领域备受关注,如高胆固醇血症、癌症化疗中的药物靶向以及作为组织培养系统中的脂质补充剂。然而,目前它只能通过从新鲜血浆样本中提取获得,产量很低。合成低密度脂蛋白(sLDL)是利用易于获得的脂质成分与含有载脂蛋白B受体序列的合成两亲性肽分子制备而成。sLDL能够以胆固醇依赖的方式支持中国仓鼠卵巢(CHO)细胞和成纤维细胞在无血清培养基中生长,这种方式与受体肽分子的存在有关。sLDL可用高氯酸3,3'-二辛基氧杂羰花青(DiO)进行荧光标记,一旦标记,CHO细胞和成纤维细胞会以时间和温度依赖的方式摄取,这种方式取决于受体肽的存在。此外,过量的未标记天然LDL会减少摄取。结果表明,sLDL与CHO细胞和成纤维细胞的相互作用是通过受体依赖系统发生的,很可能是LDL细胞受体。因此,sLDL是一种有用的、易于获得的天然LDL替代品,在细胞药物靶向和无血清组织培养系统领域具有潜在应用价值。