Department of Preventive Medicine, School of Public Health, Fujian Medical University, Fuzhou 350122, China; The Key Laboratory of Environment and Health, School of Public Health, Fujian Medical University, Fuzhou 350122, China; Department of Nutrition and Food Hygiene, Faculty of Naval Medicine, The Second Military Medical University, Shanghai 200433, China.
Department of Preventive Medicine, School of Public Health, Fujian Medical University, Fuzhou 350122, China; The Key Laboratory of Environment and Health, School of Public Health, Fujian Medical University, Fuzhou 350122, China; Fujian Provincial Key Laboratory of Environmental Factors and Cancer, School of Public Health, Fujian Medical University, Fuzhou 350122, China.
Ecotoxicol Environ Saf. 2021 Jan 15;208:111424. doi: 10.1016/j.ecoenv.2020.111424. Epub 2020 Oct 22.
Emerging evidences having suggested that particular lncRNAs have a potential effect on PD progression through provoking damage and inflammatory responses of microglia/ dopaminergic cells. In addition, paraquat can be accumulated in human body through various approaches and have an increased risk for Parkinson's disease. However, the specific role and mechanism of lncRNA related to neurotoxic in the progression of PD is unclear. In our study, a mouse PD model was established induced by the intraperitoneal injection of paraquat (5 mg/kg and 10 mg/kg) every three days (10 times). We determined differential expression of lncRNA AK039862 and its potential targeted genes Pafah1b1/Foxa1 in PD mouse model, then we used fluorescence in situ hybridization (FISH) to visualize the cellular distribution of AK039862. Short interfering RNAs (siRNAs) and overexpression plasmids were designed for knockdown or overexpression of AK039862. To simulate the coexisting dopaminergic cells and microglia cells in vitro, we applied several non-contact co-culture models, including conditioned medium and Transwell co-culture systems. Cytotoxicity of PQ was evaluated using bv2 cells with the concentrations: 30, 60 μM, and mn9d cells with the concentrations: 50, 100 μM. As a result, we depicted multiple interesting individual and interactive features of inflammatory lncRNA AK039862 involved in PQ-induced cellular functional effects. First, we detected that AK039862 contributed to the neuronal injury process in PQ-treated mice and co-localization of AK039862 with dopaminergic cells in vivo. And interestingly, we demonstrated that PQ significantly inhibited microglia and dopaminergic cells proliferation and microglia migration in vitro. Further research indicated that the PQ-induced low expression of AK039862 rescued microglia proliferation and migration inhibition via the AK039862/Pafah1b1/Foxa1 pathway. Meanwhile, AK039862 also participated in the interaction between microglia and dopaminergic cells with PQ treatment in non-contact co-culture models. In summary, we found that PQ inhibited the proliferation and migration of microglial cells, and elucidated AK039862 played a key role in PQ-induced neuroinflammatory damage through Pafah1b1/Foxa1. Finally, inflammatory AK039862 is involved in the complex communication between microglia and dopaminergic cells in the environment of PQ damage.
越来越多的证据表明,特定的长链非编码 RNA 通过引发小胶质细胞/多巴胺能细胞的损伤和炎症反应,对 PD 的进展有潜在影响。此外,马拉硫磷可以通过各种途径在人体内蓄积,患帕金森病的风险增加。然而,lncRNA 与神经毒性在 PD 进展中的关系尚不清楚。在我们的研究中,通过腹腔注射马拉硫磷(5mg/kg 和 10mg/kg)每三天一次(共 10 次)建立了 PD 小鼠模型。我们确定了 PD 小鼠模型中长链非编码 RNA AK039862 及其潜在靶向基因 Pafah1b1/Foxa1 的差异表达,然后使用荧光原位杂交(FISH)观察 AK039862 的细胞分布。设计了短干扰 RNA(siRNA)和过表达质粒来敲低或过表达 AK039862。为了模拟体外共存的多巴胺能细胞和小胶质细胞,我们应用了几种非接触共培养模型,包括条件培养基和 Transwell 共培养系统。用浓度为 30、60μM 的 bv2 细胞和浓度为 50、100μM 的 mn9d 细胞评估 PQ 的细胞毒性。结果表明,我们描绘了炎性 lncRNA AK039862 参与 PQ 诱导的细胞功能效应的多个有趣的个体和相互作用特征。首先,我们检测到 AK039862 有助于 PQ 处理小鼠的神经元损伤过程,并在体内与多巴胺能细胞共定位。有趣的是,我们证明 PQ 显著抑制体外小胶质细胞和多巴胺能细胞的增殖和小胶质细胞的迁移。进一步的研究表明,PQ 诱导的 AK039862 低表达通过 AK039862/Pafah1b1/Foxa1 通路挽救小胶质细胞增殖和迁移抑制。同时,AK039862 也参与了非接触共培养模型中 PQ 处理时小胶质细胞与多巴胺能细胞之间的相互作用。总之,我们发现 PQ 抑制小胶质细胞的增殖和迁移,并阐明 AK039862 通过 Pafah1b1/Foxa1 在 PQ 诱导的神经炎症损伤中发挥关键作用。最后,炎性 AK039862 参与了 PQ 损伤环境中小胶质细胞和多巴胺能细胞之间的复杂通讯。