Zhu Yan, Liu Zhan, Peng Yu-Ping, Qiu Yi-Hua
Department of Physiology, School of Medicine, and Co-innovation Center of Neuroregeneration, Nantong University, 19 Qixiu Road, Nantong, Jiangsu Province 226001, China.
Department of Physiology, School of Medicine, and Co-innovation Center of Neuroregeneration, Nantong University, 19 Qixiu Road, Nantong, Jiangsu Province 226001, China.
Int Immunopharmacol. 2017 Sep;50:353-360. doi: 10.1016/j.intimp.2017.07.017. Epub 2017 Jul 25.
Neuroinflammation plays an important role in the pathogenesis of Parkinson's disease. Interleukin (IL)-10 is one of the most important and best anti-inflammatory cytokines. The objective of this report is to investigate whether IL-10 has any role in protecting ventral mesencephalic (VM) neurons in in vitro model of neuroinflammation. In this study, primary neuron-enriched culture was prepared from the VM tissues of E14 embryos of rats. The cells were pretreated with IL-10 (15 or 50ng/mL) for 1h followed by lipopolysaccharide (LPS, 50ng/mL) application. We found LPS induced neuronal apoptosis and loss while pretreatment with IL-10 reduced neuronal damage after exposure of LPS toxicity. Furthermore, signal transduction pathways related to IL-10 in VM neurons were studied in inflammatory condition. We used both shRNA and pharmacologic inhibition to determine the role of the IL-10 receptor (IL-10R) and its downstream signaling pathways in LPS-induced VM neuronal toxicity. Silence of the IL-10R gene in VM neurons abolished IL-10 mediated protection and the properties of anti-inflammatory and anti-apoptosis. IL-10 also induced phosphorylation of signal transducer and activator of transcription (STAT) 3 in VM neurons. Pretreatment with the specific Janus kinase (JAK) inhibitor reduced STAT3 phosphorylation and blocked IL-10 mediated protection against LPS. These findings suggest that IL-10 provides neuroprotection by acting via IL-10R and its down-stream JAK-STAT3 signal pathways in VM neurons.
神经炎症在帕金森病的发病机制中起重要作用。白细胞介素（IL）-10是最重要且最有效的抗炎细胞因子之一。本报告的目的是研究在神经炎症的体外模型中IL-10是否对保护中脑腹侧（VM）神经元有作用。在本研究中，从大鼠E14胚胎的VM组织制备了富含原代神经元的培养物。细胞先用IL-10（15或50ng/mL）预处理1小时，然后施加脂多糖（LPS，50ng/mL）。我们发现LPS诱导神经元凋亡和损失，而用IL-10预处理可减少LPS毒性暴露后的神经元损伤。此外，在炎症条件下研究了VM神经元中与IL-10相关的信号转导途径。我们使用短发夹RNA（shRNA）和药物抑制来确定IL-10受体（IL-10R）及其下游信号通路在LPS诱导的VM神经元毒性中的作用。VM神经元中IL-10R基因的沉默消除了IL-10介导的保护作用以及抗炎和抗凋亡特性。IL-10还诱导VM神经元中信号转导和转录激活因子（STAT）3的磷酸化。用特异性Janus激酶（JAK）抑制剂预处理可降低STAT3磷酸化并阻断IL-10介导的对LPS的保护作用。这些发现表明，IL-10通过在VM神经元中经由IL-10R及其下游的JAK-STAT3信号通路发挥作用来提供神经保护。
