Jiang Shiyu, Zhang Xiang, Xu Jianghui, Liu Yi, Chen Wei, Zhang Jun, Wang Jing
Department of Lymphoma, Fudan University Shanghai Cancer Center, Fudan, China.
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
Endocr Metab Immune Disord Drug Targets. 2024 Nov 29. doi: 10.2174/0118715303321231240905073202.
The significance of circular RNAs (circRNAs) in diabetic complications has been established. However, their role in basal and diabetic states, as well as cognitive dysfunction, requires further investigation.
BV-2 microglial cells were exposed to high glucose (50mM) and insulin (2μM) for 48 hours. The levels of interleukin-1beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factoralpha (TNF-α) were assessed through quantitative polymerase chain reaction (qPCR), western blot, and ELISA. CircRNA and messenger RNA (mRNA) sequencing were performed, and the data were analyzed. Differentially expressed circRNAs and mRNAs were identified using qPCR. The circRNA-miRNA interaction was predicted using Miranda and TargetScan software, and their levels were quantified by qPCR.
The results demonstrated a significant increase in mRNA and protein levels of IL-1β, IL-6, and TNF-α in BV2 cells treated with glucose and insulin. Five circRNAs (four upregulated and one downregulated) were identified in both glucose and insulin groups compared to the control. Further qPCR analysis revealed marked increases in the levels of chr17:40159331- 40159711+ and chr2:72800499-72801858- (mmu_circ_0010164) in both treatment groups. Competitive endogenous RNA networks showed significant upregulation of mRNA levels of mitochondrial transcription termination factor 1b (Mterf1b) and G protein subunit gamma 4 (Gng4), accompanied by a decrease in mmu-miR-6918-3p and mmu-miR-7043-3p levels in the glucose and insulin groups compared to the control. Knockdown of mmu_circ_0010164 significantly inhibited the inflammatory response induced by glucose and insulin in BV-2 microglial cells.
These findings indicate that both glucose and insulin can elicit inflammatory responses in BV2 cells through the modulation of mmu_circ_0010164 levels. The underlying mechanism may involve potential downstream targets of mmu_circ_0010164, specifically mmu-miR-7043-3p/Gng4 and mmu-miR-6918-3p/Mterf1b. This provides novel insights into the treatment of glucose-induced neuroinflammation.
环状RNA（circRNAs）在糖尿病并发症中的重要性已得到证实。然而，它们在基础状态和糖尿病状态以及认知功能障碍中的作用仍需进一步研究。
将BV-2小胶质细胞暴露于高糖（50mM）和胰岛素（2μM）环境中48小时。通过定量聚合酶链反应（qPCR）、蛋白质免疫印迹法和酶联免疫吸附测定（ELISA）评估白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的水平。进行环状RNA和信使核糖核酸（mRNA）测序，并对数据进行分析。使用qPCR鉴定差异表达的环状RNA和mRNA。使用Miranda和TargetScan软件预测环状RNA-微小RNA（miRNA）相互作用，并通过qPCR对其水平进行定量。
结果表明，用葡萄糖和胰岛素处理的BV2细胞中，IL-1β、IL-6和TNF-α的mRNA和蛋白质水平显著升高。与对照组相比，在葡萄糖组和胰岛素组中均鉴定出5种环状RNA（4种上调，1种下调）。进一步的qPCR分析显示，两个处理组中chr17:40159331-40159711+和chr2:72800499-72801858-（mmu_circ_0010164）的水平显著升高。竞争性内源性RNA网络显示，与对照组相比，葡萄糖组和胰岛素组中线粒体转录终止因子1b（Mterf1b）和G蛋白亚基γ4（Gng4）的mRNA水平显著上调，同时mmu-miR-6918-3p和mmu-miR-7043-3p水平降低。敲低mmu_circ_0010164可显著抑制葡萄糖和胰岛素诱导的BV-2小胶质细胞炎症反应。
这些发现表明，葡萄糖和胰岛素均可通过调节mmu_circ_0010164水平在BV2细胞中引发炎症反应。潜在机制可能涉及mmu_circ_0010164的潜在下游靶点，特别是mmu-miR-7043-3p/Gng4和mmu-miR-6918-3p/Mterf1b。这为葡萄糖诱导的神经炎症治疗提供了新的见解。
