PDF(Pubmed)
Abstract：
Telomerase activity is directly affected by the laminin receptor precursor (LRP) protein, a highly conserved nonintegrin transmembrane receptor, which has been shown to have therapeutic effects in ageing, and age-related diseases. Recently, it has been found that overexpression of LRP-FLAG, by plasmid transfection, leads to a significant increase in telomerase activity in cell culture models. This may indicate that upregulation of LRP can be used to treat various age-related diseases. However, transfection is not a viable treatment strategy for patients. Therefore, we present a nanoencapsulated protein-based drug synthesised using poly(lactic-co-glycolic acid) (PLGA) nanocapsules for delivery of the 37 kDa LRP protein therapeutic. PLGA nanocapsules were synthesised using the double emulsification-solvent evaporation technique. Different purification methods, including filtration and centrifugation, were tested to ensure that the nanocapsules were within the optimal size range, and the BCA assay was used to determine encapsulation efficiency. The completed drug was tested in a HEK-293 cell culture model, to investigate the effect on cell viability, LRP protein levels and telomerase activity. A significant increase in total LRP protein levels with a concomitant increase in cell viability and telomerase activity was observed. Due to the observed increase in telomerase activity, this approach could represent a safer alternative to plasmid transfection for the treatment of age-related diseases.
摘要：
层粘连蛋白受体前体(LRP)蛋白直接影响端粒酶活性,一种高度保守的非整合素跨膜受体，已经证明对衰老有治疗作用，与年龄有关的疾病。最近,已经发现LRP-FLAG的过表达，通过质粒转染，导致细胞培养模型中端粒酶活性的显着增加。这可能表明LRP的上调可用于治疗各种与年龄相关的疾病。然而,转染对患者来说不是可行的治疗策略。因此,我们提供了一种纳米封装的基于蛋白质的药物，该药物使用聚（乳酸-乙醇酸）（PLGA）纳米胶囊合成，用于递送37kDaLRP蛋白治疗剂。使用双乳化-溶剂蒸发技术合成PLGA纳米胶囊。不同的纯化方法，包括过滤和离心，进行了测试，以确保纳米胶囊在最佳尺寸范围内，和BCA测定用于确定包封效率。完成的药物在HEK-293细胞培养模型中进行了测试，为了研究对细胞活力的影响，LRP蛋白水平和端粒酶活性。观察到总LRP蛋白水平的显着增加，同时细胞活力和端粒酶活性也随之增加。由于观察到端粒酶活性的增加，这种方法可能是质粒转染治疗年龄相关疾病的一种更安全的替代方法.
