Long non-coding RNA NEAT1 contributes to lipopolysaccharide-induced inflammation and apoptosis of human middle ear epithelial cells via regulating the miR-301b-3p/TLR4 axis.
Long non-coding RNA NEAT1 contributes to lipopolysaccharide-induced inflammation and apoptosis of human middle ear epithelial cells via regulating the miR-301b-3p/TLR4 axis.
复制标题
DOI：
10.3892/etm.2021.10795
复制
发表时间：
2021-12
影响因子：
2.7
通讯作者：
Long R
中科院分区：
文献类型：
--
作者：
Liu Z;Lu T;Liu S;Zhang F;Yang J;Dai S;Ruan B;Long R
急性中耳炎（AOM）是一种常见的儿童感染性疾病，伴有中耳炎和感染的体征和症状。已有研究表明，长链非编码RNA核富集丰富转录本1（NEAT 1）参与多种炎症反应，并发挥重要的调控作用。本研究的重点是NEAT 1在脂多糖（LPS）诱导的人中耳上皮细胞（HMEECs）中的生物学功能及其分子机制。逆转录-定量PCR检测NEAT 1、miR-301 b-3 p和TLR 4蛋白表达。进行双荧光素酶报告基因测定以研究miR-301 b-3 p与NEAT 1或TLR 4的组合。采用流式细胞仪检测细胞活力，ELISA法检测细胞凋亡，流式细胞仪检测促炎因子IL-1β、TNF-α和IL-6水平。细胞活力显着降低，而细胞凋亡和炎症增加，在LPS刺激的HMEEC。功能分析表明，在LPS处理后，NEAT 1被上调，而NEAT 1的敲低显著增加细胞活力，减轻细胞凋亡和炎症。从机制上讲，NEAT 1直接结合并负调控miR-301 b-3 p表达，而miR-301 b-3 p抑制剂消除了NEAT 1敲低对细胞凋亡和炎症的抑制作用。TLR 4作为miR-301 b-3 p的靶点，受NEAT 1和miR-301 b-3 p的调控。TLR 4过表达减轻NEAT 1沉默诱导的炎症抑制。拯救实验表明NEAT 1通过抑制miR-301 b-3 p促进TLR 4表达。总之，本研究结果提示NEAT 1可能通过调节miR-301 b-3 p/TLR 4轴来减轻LPS诱导的HMEECs炎症和凋亡，并可能为AOM的临床治疗提供新的治疗靶点。
Acute otitis media (AOM) is a common infectious disease in children that is accompanied by signs and symptoms of middle ear inflammation and infection. Previous studies have shown that the long non-coding (lnc)RNA nuclear-enriched abundant transcript 1(NEAT1) participates in various inflammatory conditions and plays an important regulatory role. The focus of the present study was the biological function of NEAT1 and underlying molecular mechanism in lipopolysaccharide (LPS)-induced human middle ear epithelial cells (HMEECs). The expression of NEAT1, miR-301b-3p and toll-like receptor 4 (TLR4) protein were determined by reverse transcription-quantitative PCR and western blot assays, respectively. Dual-luciferase reporter assay was performed to investigate the combination of miR-301b-3p and NEAT1 or TLR4. In addition, cell viability, apoptosis and the levels of pro-inflammatory factors (IL-1β, TNF-α and IL-6) were measured by Cell Counting Kit-8 assay, flow cytometry and ELISA, respectively. Cell viability was significantly decreased, whereas apoptosis and inflammation were increased in LPS-stimulated HMEECs. Functional analyses demonstrated that NEAT1 was upregulated following LPS treatment, whereas knockdown of NEAT1 significantly increased cell viability and alleviated apoptosis and inflammation. Mechanistically, NEAT1 directly bound to and negatively regulated miR-301b-3p expression, whereas miR-301b-3p inhibitors abolished the inhibitory effect of NEAT1 knockdown on cell apoptosis and inflammation. As a target of miR-301b-3p, TLR4 was regulated by NEAT1 and miR-301b-3p. TLR4 overexpression alleviated NEAT1 silencing-induced inflammatory suppression. Rescue experiments demonstrated that NEAT1 promoted TLR4 expression by inhibiting miR-301b-3p. Collectively, the results of the present study suggested that NEAT1 may attenuate LPS-induced inflammation and apoptosis in HMEECs by modulating the miR-301b-3p/TLR4 axis, and may provide a new therapeutic target for the clinical treatment of AOM.
登录
查看更多内容
影响因子：
64.5
作者：
Gomez JA;Wapinski OL;Yang YW;Bureau JF;Gopinath S;Monack DM;Chang HY;Brahic M;Kirkegaard K
通讯作者：
Kirkegaard K
影响因子：
28.2
作者：
Karreth FA;Pandolfi PP
通讯作者：
Pandolfi PP
影响因子：
7.5
作者：
Guo, Hao;Li, Ming;Xu, Li-Juan
通讯作者：
Xu, Li-Juan
DOI：
10.26355/eurrev_202002_20195
发表时间：
2020-01-01
影响因子：
3.3
作者：
Chen, X-X;Jiang, Y. J.;Li, J-J
通讯作者：
Li, J-J
坂崎克罗诺杆菌通过 TLR4 调节 NLRP3 炎性体表达，诱导坏死性小肠结肠炎。
影响因子：
3
作者：
Chen, Zhenhui;Zhang, Yiduo;Fan, Hongying
通讯作者：
Fan, Hongying