Preparation of Biodegradable PLGA-Nanoparticles Used for pH-Sensitive Intracellular Delivery of an Anti-inflammatory Bacterial Toxin to Macrophages
Preparation of Biodegradable PLGA-Nanoparticles Used for pH-Sensitive Intracellular Delivery of an Anti-inflammatory Bacterial Toxin to Macrophages
复制标题
DOI：
10.1248/cpb.c19-00917
复制
发表时间：
2020-04-01
影响因子：
1.7
通讯作者：
Niidome, Takuro
中科院分区：
文献类型：
--
作者：
Harada, Ayaka;Tsutsuki, Hiroyasu;Niidome, Takuro
聚（D，L-丙交酯-共-乙醇酸）（PLGA）是一种合成共聚物，已用于设计微米/纳米颗粒作为蛋白质和核酸等大分子的载体，这些大分子可以通过内吞途径内化。然而，很难控制细胞内递送到靶细胞器。在这里，我们报告了一个细胞内传递系统的纳米粒子修饰的细菌细胞毒素的内质网（ER）和抗炎活性的纳米粒子。枯草杆菌酶细胞毒素（SubAB）是某些肠出血性大肠杆菌（EHEC）菌株中的细菌毒素，其在亚致死浓度下切割宿主ER伴侣BiP并抑制巨噬细胞中的核因子-κ B（NF-κ B）活化和一氧化氮（NO）产生。通过pH敏感性连接，用寡聚组氨酸标记的（6 x His标记的）重组SubAB（SubAB-PLGA）修饰PLGA纳米颗粒，并检查其在巨噬细胞系J774.1细胞中向ER的易位、对诱导型NO合酶（iNOS）的影响以及由脂多糖（LPS）诱导的肿瘤坏死因子（TNF）-α细胞因子水平。与游离SubAB相比，SubAB-PLGA在J774.1细胞中的BiP切割和ER应激标志物C/EBP同源蛋白（CHOP）的诱导中显著有效。此外，SubAB-PLGA减弱LPS刺激的iNOS和TNF-α诱导。我们的研究结果为蛋白质递送到巨噬细胞提供了有用的信息，并可能鼓励纳米颗粒治疗炎症性疾病的治疗应用。
Poly(D,L-lactide-co-glycolic) acid (PLGA) is a synthetic copolymer that has been used to design micro/nanoparticles as a carrier for macromolecules, such as protein and nucleic acids, that can be internalized by the endocytosis pathway. However, it is difficult to control the intracellular delivery to target organelles. Here we report an intracellular delivery system of nanoparticles modified with bacterial cytotoxins to the endoplasmic reticulum (ER) and anti-inflammatory activity of the nanoparticles. Subtilase cytotoxin (SubAB) is a bacterial toxin in certain enterohemorrhagic Escherichia coli (EHEC) strains that cleaves the host ER chaperone BiP and suppresses nuclear factor-kappaB (NF-kappa B) activation and nitric oxide (NO) generation in macrophages at sub-lethal concentration. PLGA-nanoparticles were modified with oligo histidine-tagged (6 x His-tagged) recombinant SubAB (SubAB-PLGA) through a pH-sensitive linkage, and their translocation to the ER in macrophage cell line J774.1 cells, effects on inducible NO synthase (iNOS), and levels of tumor necrosis factor (TNF)-alpha cytokine induced by lipopolysaccharide (LPS) were examined. Compared with free SubAB, SubAB-PLGA was significantly effective in BiP cleavage and the induction of the ER stress marker C/EBP homologous protein (CHOP) in J774.1 cells. Furthermore, SubAB-PLGA attenuated LPS-stimulated induction of iNOS and TNF-alpha. Our findings provide useful information for protein delivery to macrophages and may encourage therapeutic applications of nanoparticles to the treatment of inflammatory diseases.