Department of Neurology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China.
Department of Neurology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China.
Int J Pharm. 2019 Aug 15;567:118449. doi: 10.1016/j.ijpharm.2019.118449. Epub 2019 Jun 18.
In the present study, we have prepared microRNA(miR)-124-loaded Rabies virus glycoprotein (RVG)29 surface-conjugated polymeric nanoparticles (NPs) to improve neuroinflammation in Parkinson's disease (PD). We hypothesize that an increase in the intracellular concentration of miR-124 will result in a better prognosis for Parkinson's disease. Minimal toxicity for the RVG29 NPs was observed at concentrations <100 µg/mL, while the cell viability of cells treated with blank NPs at concentrations of 200 µg/mL markedly decreased, indicating the safety of the carrier system. Results showed that mRNA levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-α), and interleukin (IL)-6, significantly increased upon lipopolysaccharide (LPS) administration. However, the mRNA levels of these cytokines reflected those of the miR-NPs-treated control group, indicating the influence of miR-124 exposure. After transfection with miR-NPs, levels of pro-inflammatory cytokines and neuroprotective molecules were reduced and increased, respectively. Administration of LPS significantly increased the levels of mitogen activated protein kinase kinase kinase (MEKK)3 and P-P65 levels, while transfection with miR-NPs significantly reduced the expression of both MEKK3 and P-P65, reflecting that of the control. This research has revealed that miR-124 could target both the MEKK3 and nuclear factor kappa light chain enhancer of activated B cell (NF-Kb) pathways, while also reducing inflammatory cytokine levels. In addition, a 3-fold decrease in apoptosis was observed in miR-NP transfected cells. The exogenous delivery of miR-NPs significantly downregulated MEKK3 expression in animal studies, as outlined by immunohistochemical staining (IHC). Overall, miR-NPs have the potential to inhibit pro-inflammatory signaling and enhance neuroprotection in PD.
在本研究中,我们制备了装载 microRNA(miR)-124 的狂犬病病毒糖蛋白 (RVG)29 表面偶联聚合物纳米颗粒 (NPs),以改善帕金森病 (PD) 中的神经炎症。我们假设细胞内 miR-124 浓度的增加将导致帕金森病的预后更好。在浓度<100μg/mL 时,RVG29 NPs 的毒性最小,而空白 NPs 处理的细胞在浓度为 200μg/mL 时的细胞活力明显下降,表明载体系统的安全性。结果表明,在脂多糖 (LPS) 给药后,诱导型一氧化氮合酶 (iNOS)、肿瘤坏死因子-α (TNF-α) 和白细胞介素 (IL)-6 的 mRNA 水平显著增加。然而,这些细胞因子的 mRNA 水平反映了 miR-NPs 处理对照组的水平,表明 miR-124 暴露的影响。转染 miR-NPs 后,促炎细胞因子和神经保护分子的水平分别降低和增加。LPS 给药显著增加丝裂原激活蛋白激酶激酶激酶 (MEKK)3 和 P-P65 水平,而转染 miR-NPs 则显著降低 MEKK3 和 P-P65 的表达,反映了对照组的情况。这项研究表明,miR-124 可以靶向 MEKK3 和核因子 kappa 轻链增强子的激活 B 细胞 (NF-Kb) 途径,同时降低炎症细胞因子水平。此外,在 miR-NP 转染的细胞中观察到凋亡减少了 3 倍。免疫组织化学染色 (IHC) 表明,外源性递送 miR-NPs 可显著下调动物研究中 MEKK3 的表达。总的来说,miR-NPs 有可能抑制 PD 中的促炎信号并增强神经保护作用。