Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, National Clinical Research Center for Child Health and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children's Hospital of Chongqing Medical University, Chongqing, China.
Institute of Aging, Key Laboratory of Alzheimer's Disease of Zhejiang Province, Zhejiang Provincial Clinical Research Center for Mental Disorders, School of Mental Health and Kangning Hospital, The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
J Alzheimers Dis. 2022;88(4):1469-1485. doi: 10.3233/JAD-220321.
Interleukin-10 (IL-10) is a classic anti-inflammatory cytokine that exerts its effects via the receptor complexes IL-10RA and IL-10RB. Loss of IL-10RB results in many diseases. Moreover, IL-10RB is closely associated with neuronal survival and synaptic formation. However, the regulation of IL-10RB gene expression remains elusive.
To investigate whether the expression of IL-10RB gene is increased in brain of Alzheimer's disease (AD) and its transcriptional regulation.
We examined the gene expression of AD patient brain from public database and detected the protein expression of AD model mouse brain by western blot. We constructed a variety of reporter gene plasmids with different lengths or mutation sites, tested the promoter activity and defined the functional region of the promoter with the luciferase reporter assay. The protein-DNA binding between transcription factors and the promoter was analyzed using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA).
We found that the IL-10RB is elevated in the brain of AD patient and AD model mice. The minimal promoter of the IL-10RB gene is located in the -90 to +51 bp region (relative to the transcriptional start site) and is sufficient for high-level expression of the IL-10RB gene. Transcription factors Sp8 and Sp9 bind to the IL-10RB promoter in vitro. The overexpression or knockdown of Sp8 and Sp9 affected the IL-10RB promoter activity and its gene expression.
Our study functionally characterized the promoter of the IL-10RB gene and demonstrated that Sp8 and Sp9 regulated its expression.
白细胞介素-10（IL-10）是一种经典的抗炎细胞因子，通过受体复合物 IL-10RA 和 IL-10RB 发挥作用。IL-10RB 的缺失会导致多种疾病。此外，IL-10RB 与神经元存活和突触形成密切相关。然而，IL-10RB 基因表达的调控仍不清楚。
研究阿尔茨海默病（AD）患者大脑中 IL-10RB 基因的表达是否增加及其转录调控。
我们从公共数据库中检查了 AD 患者大脑的基因表达，并通过 Western blot 检测了 AD 模型小鼠大脑的蛋白质表达。我们构建了具有不同长度或突变位点的各种报告基因质粒，通过荧光素酶报告基因检测法测试启动子活性并确定启动子的功能区域。使用染色质免疫沉淀（ChIP）和电泳迁移率变动分析（EMSA）分析转录因子与启动子之间的蛋白-DNA 结合。
我们发现 AD 患者和 AD 模型小鼠大脑中 IL-10RB 升高。IL-10RB 基因的最小启动子位于-90 至+51 bp 区域（相对于转录起始位点），足以高水平表达 IL-10RB 基因。转录因子 Sp8 和 Sp9 在体外与 IL-10RB 启动子结合。Sp8 和 Sp9 的过表达或敲低会影响 IL-10RB 启动子活性及其基因表达。
我们的研究功能表征了 IL-10RB 基因的启动子，并证明 Sp8 和 Sp9 调节其表达。
