Jebali A, Rashidi M, Keikha R, Daliri K, Outeiro T F
Department of Medical Nanotechnology, Faculty of Advanced Sciences and Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
Department of Pharmacology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran; The Health of Plant and Livestock Products Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Neurologia (Engl Ed). 2024 May;39(4):321-328. doi: 10.1016/j.nrleng.2021.08.003.
The aim of this study was to compare the effect of five types of PEGlated nanoliposomes (PNLs) on α-synuclein (α-syn) fibrillization, attenuation of microglial activation, and silence of the SNCA gene, which encodes α-syn.
To evaluate the inhibition of α-syn fibrillization, we used standard in vitro assay based on Thioflavin T (ThT) fluorescence. Next, to evaluate the attenuation of microglial activation, the concentration of TNF-a and IL-6 was quantified by ELISA assay in BV2 microglia cells treated with 100nM A53T α-syn and PNLs. In order to determine the silencing of the SNCA, real-time PCR and Western blot analysis was used. Finally, the efficacy of PNLs was confirmed in a transgenic mouse model expressing human α-syn.
ThT assay showed both PNL1 and PNL2 significantly inhibited a-syn fibrillization. ELISA test also showed the production of TNF-a and IL-6 was significantly attenuated when microglial cells treated with PNL1 or PNL2. We also found that SNCA gene, at both mRNA and protein levels, was significantly silenced when BV2 microglia cells were treated with PNL1 or PNL2. Importantly, the efficacy of PNL1 and PNL2 was finally confirmed in vivo in a transgenic mouse model.
In conclusion, the novel multifunctional nanoliposomes tested in our study inhibit α-syn fibrillization, attenuate microglial activation, and silence SNCA gene. Our findings suggest the therapeutic potential of PNL1 and PNL2 for treating synucleinopathies.
本研究的目的是比较五种聚乙二醇化纳米脂质体（PNLs）对α-突触核蛋白（α-syn）纤维化、小胶质细胞激活的减弱以及编码α-syn的SNCA基因沉默的影响。
为了评估对α-syn纤维化的抑制作用，我们使用了基于硫黄素T（ThT）荧光的标准体外试验。接下来，为了评估小胶质细胞激活的减弱情况，在用100nM A53T α-syn和PNLs处理的BV2小胶质细胞中，通过ELISA测定法对TNF-α和IL-6的浓度进行定量。为了确定SNCA的沉默情况，使用了实时PCR和蛋白质印迹分析。最后，在表达人α-syn的转基因小鼠模型中证实了PNLs的疗效。
ThT试验表明PNL1和PNL2均显著抑制α-syn纤维化。ELISA试验还表明，当用PNL1或PNL2处理小胶质细胞时，TNF-α和IL-6的产生显著减弱。我们还发现，当BV2小胶质细胞用PNL1或PNL2处理时，SNCA基因在mRNA和蛋白质水平均显著沉默。重要的是，最终在转基因小鼠模型的体内证实了PNL1和PNL2的疗效。
总之，我们研究中测试的新型多功能纳米脂质体抑制α-syn纤维化，减弱小胶质细胞激活，并使SNCA基因沉默。我们的研究结果表明PNL1和PNL2在治疗突触核蛋白病方面具有治疗潜力。
