Background

Ischemic stroke is one of the major diseases of the cerebral vasculature. Currently, Ischemic stroke is the leading cause of neurological disability worldwide and has a high morbidity and mortality rate. The NF-κB interacting lncRNA (NKILA), the recently identified, is a key booster of NF-κB pathway. Accumulating studies have shown that NKILA plays a cancer suppressor in a variety of malignancies by regulating the NF-κB pathway. Nevertheless, the role of NKILA in ischemic stroke remains to be elucidated.

Methods

We constructed a mouse model of middle cerebral artery occlusion-reperfusion (MCAO/R). TTC staining and dry and wet weight method were used to evaluate infarction and water content of brain tissue. RT-qPCR was performed to detect NKILA expression in cerebral infarction tissues. After labeling astrocytes and neurons with GFAP and NeuN, respectively, EDU and TUNEL staining were performed. Inflammatory factor levels were detected by ELISA. Commercial kits were used to detect the levels of oxidative stress-related factors. In in vitro, the HT22/U251 cell co-culture model was used for oxygen-glucose deprivation and re-introduction (OGD/R) to verify the effect of NKILA on neuronal cell inflammation and oxidative stress through astrocytes.

Results

In in vivo experiments, NKILA significantly reduced cerebral infarction volume, brain water content and neurological score caused by MCAO/R. Moreover, NKILA blocked the activation of the NF-κB pathway, and inhibited astrocyte proliferation and neuron apoptosis as well as inflammation and oxidative stress. In in vitro experiments, NKILA significantly inhibited NF-κB pathway in HT22 cells. In addition, NKILA alleviated the inflammatory response and oxidative stress of U251 cells mediated by HT22 cells after OGD/R, and promoted U251 cell proliferation and inhibit their apoptosis.

Conclusions

In summary, we found for the first time that NKILA alleviates inflammatory response and oxidative stress after cerebral ischemia/reperfusion by blocking the activation of NF-κB pathway.

中文翻译：

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LncRNA NKILA通过抑制NF-κB通路缓解脑缺血/再灌注后星形胶质细胞炎症和神经元氧化应激

背景

缺血性中风是脑血管系统的主要疾病之一。目前，缺血性中风是全球神经功能障碍的主要原因，并且具有很高的发病率和死亡率。最近发现的 NF-κB 相互作用的 lncRNA (NKILA) 是 NF-κB 通路的关键助推器。越来越多的研究表明，NKILA 通过调节 NF-κB 通路在多种恶性肿瘤中发挥抑癌作用。然而，NKILA 在缺血性卒中中的作用仍有待阐明。

方法

我们构建了大脑中动脉闭塞再灌注（MCAO/R）的小鼠模型。采用TTC染色和干湿重法评价脑组织梗死和含水量。进行RT-qPCR以检测脑梗死组织中NKILA的表达。分别用 GFAP 和 NeuN 标记星形胶质细胞和神经元后，进行 EDU 和 TUNEL 染色。通过ELISA检测炎症因子水平。商业试剂盒用于检测氧化应激相关因子的水平。在体外， HT22/U251细胞共培养模型用于氧-葡萄糖剥夺和重新引入（OGD/R），以验证NKILA通过星形胶质细胞对神经元细胞炎症和氧化应激的影响。

结果

在体内实验中，NKILA显着降低了MCAO/R引起的脑梗死体积、脑含水量和神经系统评分。此外，NKILA 阻断 NF-κB 通路的激活，并抑制星形胶质细胞增殖和神经元凋亡以及炎症和氧化应激。在体外实验中，NKILA显着抑制HT22细胞中的NF-κB通路。此外，NKILA减轻了OGD/R后HT22细胞介导的U251细胞的炎症反应和氧化应激，促进U251细胞增殖并抑制其凋亡。

结论

总之，我们首次发现NKILA通过阻断NF-κB通路的激活来减轻脑缺血/再灌注后的炎症反应和氧化应激。