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在脂多糖诱导的脓毒症细胞模型中,miR-1184通过靶向TRADD来调节炎症反应和细胞凋亡。

miR-1184 regulates inflammatory responses and cell apoptosis by targeting TRADD in an LPS-induced cell model of sepsis.

作者信息

Ling Ping, Tang Rong, Wang Huazhu, Deng Xiuqin, Chen Jianli

机构信息

Pediatric Intensive Care Unit, Guiyang Maternal and Child Health Care Hospital, Guiyang, Guizhou 550003, P.R. China.

出版信息

Exp Ther Med. 2021 Jun;21(6):630. doi: 10.3892/etm.2021.10062. Epub 2021 Apr 15.

Abstract

MicroRNAs (miRs) have been reported to be potential clinical biomarkers for sepsis. miR-1184 is a multifunctional microRNA that exerts roles in the development of various diseases. However, the role of miR-1184 in children with sepsis remain unknown. In the present study, THP-1 cells were stimulated with 1 µg/ml lipopolysaccharide (LPS) for 24 h to establish an sepsis model. Reverse transcription-quantitative PCR was used to evaluate the expression of miR-1184 in clinical specimens, and of IL-6, TNF-α, IL-1β, miR-1184 and TNF receptor type 1-associated DEATH domain protein (TRADD) in cells with and without LPS treatment. Cell apoptosis was assessed using flow cytometry. Binding between miR-1184 and TRADD was predicted using bioinformatics software, and a luciferase reporter assay was performed to verify the interaction between miR-1184 and TRADD in LPS-induced THP-1 cells. In addition, western blot analysis was performed to detect TRADD and proteins associated with the NF-κB pathway. The results showed that miR-1184 was downregulated in the blood of children with sepsis and LPS-induced THP-1 cells. Overexpression of miR-1184 alleviated the LPS-induced production of inflammatory cytokines and cell apoptosis. Moreover, TRADD was verified to be a direct target of miR-1184. Upregulation of TRADD reversed the effects of miR-1184 on the LPS-induced inflammatory response and apoptosis of THP-1 cells. Furthermore, the NF-κB pathway was shown to be associated with the regulatory role of miR-1184 in sepsis. The present study provides evidence that miR-1184 exerts inhibitory effects on inflammatory responses and apoptosis in sepsis by targeting TRADD, which suggests that miR-1184 may be a novel potential target for the therapy of children with sepsis.

摘要

据报道,微小RNA(miR)是脓毒症潜在的临床生物标志物。miR-1184是一种多功能微小RNA,在多种疾病的发展过程中发挥作用。然而,miR-1184在脓毒症患儿中的作用尚不清楚。在本研究中,用1μg/ml脂多糖(LPS)刺激THP-1细胞24小时以建立脓毒症模型。采用逆转录定量PCR评估临床标本中miR-1184的表达,以及LPS处理和未处理细胞中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、miR-1184和I型肿瘤坏死因子受体相关死亡结构域蛋白(TRADD)的表达。使用流式细胞术评估细胞凋亡。使用生物信息学软件预测miR-1184与TRADD之间的结合,并进行荧光素酶报告基因测定以验证LPS诱导的THP-1细胞中miR-1184与TRADD之间的相互作用。此外,进行蛋白质印迹分析以检测TRADD和与核因子-κB(NF-κB)途径相关的蛋白质。结果表明,脓毒症患儿血液和LPS诱导的THP-1细胞中miR-1184表达下调。miR-1184过表达减轻了LPS诱导的炎性细胞因子产生和细胞凋亡。此外,TRADD被证实是miR-1184的直接靶标。TRADD上调逆转了miR-1184对LPS诱导的THP-1细胞炎性反应和凋亡的影响。此外,NF-κB途径显示与miR-1184在脓毒症中的调节作用有关。本研究提供了证据表明miR-1184通过靶向TRADD对脓毒症中的炎性反应和凋亡发挥抑制作用,这表明miR-1184可能是脓毒症患儿治疗的新型潜在靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8bef/8082660/36e046501110/etm-21-06-10062-g00.jpg

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