College of Pharmacology, the Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Medical University, Chongqing, China.
Library, Southwest Medical University, Luzhou, Sichuan, China.
Neurol Res. 2021 Sep;43(9):767-777. doi: 10.1080/01616412.2021.1934317. Epub 2021 Jun 3.
Increasing evidence suggests that long-noncoding RNAs can exert neuroprotective effects in cerebral ischemia-reperfusion injury. Levels of the long noncoding RNA ANRIL (ANRIL) are reportedly altered in ischemic stroke (IS) patients, but its role in IS requires further clarification. This study was designed to explore the mechanistic function of ANRIL in IS.
, HT22 cells was treated with an oxygen-glucose deprivation/reperfusion (OGD/R). , brain ischemia/reperfusion was induced by 60-minute transient middle cerebral artery occlusion/reperfusion (MCAO/R) IS model in C57/BL6 mice. Additionally, cells were transfected with si-ANRIL, pcDNA3.1-ANRIL, pcDNA3.1-NF-κB, or appropriate negative controls, and si-ANRIL and pcDNA3.1-NF-κB were administered into the lateral ventricles in MCAO/R model mice. Cell viability and apoptosis were detected via MTT and flow cytometry assays. mRNA and protein expression of NF-κB were detected via qRT-PCR and Western blotting. IL-1β, IL-6, TNF-a, and iNOS levels were detected via ELISA. In addition, infarcted area and neuronal injury were evaluated via TTC, Nissl, and immunofluorescent staining.
We found that ANRIL knockdown increased cell viability and reduced apoptosis . Additionally, we found that ANRIL knockdown decreased p-P65, P65, IL-1β, IL-6, TNF-a, and iNOS levels, whereas these effects were reversed by NF-κB overexpression both and .
our results suggest that ANRIL knockdown attenuates neuroinflammation by suppressing the expression of NF-κB both and vivo model of IS, sugguesting that ANRIL might be a potentially viable therapeutictarget to diminish neuroinflammation in IS patients.
越来越多的证据表明，长非编码 RNA 可以在脑缺血再灌注损伤中发挥神经保护作用。据报道，长非编码 RNA ANRIL（ANRIL）的水平在缺血性脑卒中（IS）患者中发生改变，但它在 IS 中的作用尚需进一步阐明。本研究旨在探讨 ANRIL 在 IS 中的作用机制。
在 HT22 细胞中进行氧葡萄糖剥夺/再灌注（OGD/R）处理。在 C57/BL6 小鼠中诱导 60 分钟短暂性大脑中动脉闭塞/再灌注（MCAO/R）IS 模型，诱导脑缺血/再灌注。此外，用 si-ANRIL、pcDNA3.1-ANRIL、pcDNA3.1-NF-κB 或适当的阴性对照转染细胞，并将 si-ANRIL 和 pcDNA3.1-NF-κB 注入 MCAO/R 模型小鼠的侧脑室。通过 MTT 和流式细胞术检测细胞活力和细胞凋亡。通过 qRT-PCR 和 Western blot 检测 NF-κB 的 mRNA 和蛋白表达。通过 ELISA 检测 IL-1β、IL-6、TNF-a 和 iNOS 水平。此外，通过 TTC、Nissl 和免疫荧光染色评估梗死面积和神经元损伤。
我们发现，ANRIL 敲低可增加细胞活力并减少细胞凋亡。此外，我们发现，ANRIL 敲低可降低 p-P65、P65、IL-1β、IL-6、TNF-a 和 iNOS 水平，而 NF-κB 过表达可逆转这些作用。
我们的研究结果表明，ANRIL 敲低通过抑制 NF-κB 的表达减轻神经炎症，无论是在体内还是体外 IS 模型中，这表明 ANRIL 可能是减少 IS 患者神经炎症的潜在治疗靶点。
