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鉴定 lncRNA/circRNA-miRNA-mRNA 网络,探讨蓖麻毒素诱导 RAW264.7 细胞炎症的作用。

Identification of a lncRNA/circRNA-miRNA-mRNA network to explore the effects of ricin toxin-induced inflammation in RAW264.7 cells.

机构信息

Changchun Veterinary Research Institute, Chinese Academy of Agricultural Science, Changchun, 130122, China.

Department of Epidemiology and Biostatistics, School of Public Health, No. 1163 Xinmin Street, Jilin University, Changchun, 130021, China.

出版信息

Toxicon. 2021 Nov;203:129-138. doi: 10.1016/j.toxicon.2021.10.007. Epub 2021 Oct 18.

Abstract

Ricin toxin (RT) is a ribosome-inactivating protein derived from the beans of the castor oil plant. Our previous studies have reported that RT can induce the production of inflammatory cytokines and cause inflammatory injury in RAW264.7 cells. In order to explore the various biological processes that long noncoding RNA (lncRNA), circular RNA (circRNA) and micro RNA (miRNA) as endogenous non-coding RNAs (ceRNAs) may participate in the pro-inflammatory mechanism, RT (20 ng/mL) treated and normal RAW264.7 cells were firstly sequenced by RNA-seq. By comparing the differentially expressed genes, we obtained 10 hub genes and enriched the inflammatory-related signaling pathways. Based on our results, we concluded a lncRNA/circRNA-miRNA-mRNA network. Finally, we verified the key genes and pathways by qRT-PCR, WB and ELISA. From the experiment results, an opening MAPK signaling pathway in TNF signaling pathway via TNFR2 was found involved in RT-induced inflammation. This work provides a reference for searching for ceRNA targets or therapeutic drugs in RT-induced inflammatory injury in the future.

摘要

蓖麻毒素(RT)是一种核糖体失活蛋白,来源于巴豆植物的豆荚。我们之前的研究报告称,RT 可以诱导炎症细胞因子的产生,并在 RAW264.7 细胞中引起炎症损伤。为了探索长链非编码 RNA(lncRNA)、环状 RNA(circRNA)和 micro RNA(miRNA)等内源性非编码 RNA(ceRNA)可能参与促炎机制的各种生物学过程,我们首先通过 RNA-seq 对 RT(20ng/mL)处理和正常 RAW264.7 细胞进行了测序。通过比较差异表达基因,我们获得了 10 个枢纽基因,并富集了炎症相关的信号通路。基于我们的结果,我们构建了一个 lncRNA/circRNA-miRNA-mRNA 网络。最后,我们通过 qRT-PCR、WB 和 ELISA 验证了关键基因和通路。从实验结果中发现,RT 诱导的炎症通过 TNFR2 打开了 TNF 信号通路中的 MAPK 信号通路。这项工作为未来寻找 RT 诱导的炎症损伤中 ceRNA 靶标或治疗药物提供了参考。

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